Гиспидин-синтазы и их применение

Группа изобретений относится к области биотехнологии, конкретно к белкам биосинтеза люциферина грибов, представляющим собой гиспидин-синтазу. Предложены новые гиспидин-синтазы, кодирующие их нуклеиновые кислоты, кассеты экспрессии и клетки-хозяевы для получения указанных ферментов, применение гиспидин-синтаз в биосинтезе люциферина грибов и в способе биосинтеза предлюциферина грибов в системах in vitro и in vivo. Изобретения обеспечивают эффективный способ синтеза гиспидина и люциферина грибов и могут быть использованы для получения автономных биолюминесцентных систем, обладающих видимым свечением. 7 н. и 12 з.п. ф-лы, 9 ил., 6 табл., 25 пр.

СПОСОБ ПОЛУЧЕНИЯ ХИРАЛЬНЫХ 2-АРИЛМОРФОЛИНОВ

Изобретение относится к новому способу получения хиральных 2-(4-аминофенил)морфолинов формулы. Технический результат: разработан новый способ получения хиральных 2-(4-аминофенил)морфолинов, обладающих сродством к рецепторам, ассоциированным со следовыми аминами (TAARs), включающий стадию ферментативного восстановления кетона формулыс образованием хирального спирта формулыс последующими стадиями получения целевого продукта формулы I. 7 з.п. ф-лы, 15 пр.

СПОСОБ ПОЛУЧЕНИЯ СОДЕРЖАЩИХ ЛИГНИН ПОЛИМЕРОВ, ЛИГНИН-СОДЕРЖАЩИЕ ПОЛИМЕРЫ

Сущность изобретения: полимеры, которые содержат лигнин и органические соединения, получают тем, что лигнин полимеризуют с органическими соединениями, которые содержат по меньшей мере 3 атома углерода, а также кислород-, азотсодержащую функцию и/или связи, отличные от простой в присутствии радикально оксиляющих ферментов и окислителей, которые образуют их субстраты.

Способ получения сесквитерпенового производного

Способ получения @ -кофигурированных замещенных фенилпропанола

СПОСОБ ПОЛУЧЕНИЯ S-КОНФИГУРИРОВАННЫХ ЗАМЕЩЕННЫХ ФЕНИЛПРОПАНОЛА общей формулы Б -( Нг- clH- ciHzон. тГ ОНз где R- водород или метоксил, водород, трет-бутил, третбутоксил или метоксил отличающийся тем, что, с целью получения (-) -энантиомеров -целевых соединений, соответствующее замещенное коричного альдегида подвергают микробиологическому гидрированию с помощью дрожжей СО Saccharomyces cerevisiae : при 1040°С под воздействием аэрации возс духом в присутствии сахарозы с последуюр ей экстракцией полученного продукта хлористым метиленом и вьщелением целевого продукта упариванием. со 00 СП ...

VERBESSERTE INDOL-BIOSYNTHESE

D-N-CARBAMOYLAMINOSÄUREAMIDOHYDROLASE UND HYDANTOINASE

Enzymatisches Verfahren zur Reduktion von Oxoverbindungen, insbesondere Acetophenon und dafür geeignetes Enzym

MIKROBISCHE HERSTELLUNG VON BRENZKATECHINEN.

Fraktionierung von Allylisothiocyanat und P-Hydroxybenzylisothiocyanat aus Senf

Zur Herstellung von Allylisothiocyanat und p-Hydroxybenzylisothiocyanat kann weißer oder schwarzer Senfsamen fraktioniert werden. Es sind Verfahren zur Herstellung dieser Fraktionen aus Senfsamen-Ausgangsstoff offenbart. Der Saat-Ausgangsstoff wird mit Wasser vermischt, um eine aktivierte Suspension zu ergeben, in welcher im Saat-Ausgangsstoff vorhandene Glykosinolate durch das Enzym Myrosinase zu Isothiocyanaten hydrolysiert und abgebaut werden. Danach kann die übrig gebliebene Suspension sterilisiert und weiter auf herkömmliche Weise verarbeitet werden, um verbesserte fertige Senfprodukte zu ergeben.

Verfahren zur Herstellung von ligninhaltigen Polymeren

Mikrobielle Esterase zur enantioselektiven Spaltung von 1-Arylalkylestern

A microbial esterase enantioselectively cleaves (S)-1-phenylethyl acetate and is obtained from Arthrobacter spec. (DSM 7034), Pseudomonas fluorescens (DSM 7033) or Bacillus subtilis (DSM 7035). Also disclosed are a process for enantioselectively cleaving an 1-arylalkylester of a carboxylic acid and a process for producing an enantiomerically pure 1-arylalkynol by using this esterase.

Biotechnologische Herstellung von Cannabinoiden

Die vorliegende Erfindung betrifft ein Verfahren zur rekombinanten Herstellung von Cannbigerolsäure in einem Wirtsorganismus mit Hilfe einer modifizierten Prenyltransferase. Ferner betrifft die Erfindung eine modifizierte Prenyltransferase, ein Nukleinsäuremolekül, welches für eine solche modifizierte Prenyltransferase kodiert und einen rekombinanten Organismus, der die modifizierte Prenyltransferase und/oder das Nukleinsäuremolekül umfasst.

Method for manufacturing binder-free moulded articles

Published without abstract.

VITAMIN D3

PREPARATION OF ALCOHOLS BY ENZYME-CATALYSED REDUCTION OF ALDEHYDES

Alcohols, such as aromatic and aliphatic alcohols, are prepared by reducing the corresponding aldehyde with an alcohol dehydrogenase enzyme and with a cofactor for the enzyme. The enzyme is immobilised by covalent bonding to a support material to ameliorate the problem of its inhibition by the aldehyde and hence increase its active life. The enzyme is preferably an enzyme of Enzyme Commission class 1.1.1.1. such as yeast alcohol dehydrogenase (YADH) or horse liver alcohol dehydrogenase (HLADH). The support material may, for example, be agarose or an oxirane-acrylic polymer.

MICROBIOLOGICAL OXIDATIONS

Production method for generating 3, 5-dihydroxy-4-methoxybenzyl alcohol from oyster meat

... [Problem] The purpose of the present invention is to provide a production method with which 3,5-dihydroxy-4-methoxybenzyl alcohol, which is completely undetected in raw oyster meat, can be produced at the stage of extracting oyster meat extract. [Solution] The present invention is characterized by producing 3,5-dihydroxy-4-methoxybenzyl alcohol from heat-treated oyster meat juice by heating, for six hours or longer at 98 to 100˚C, raw oyster meat in which 3,5-dihydroxy-4-methoxybenzyl alcohol is not detected when the oyster meat is in a raw state.

Metabolically engineered cells for the production of resveratrol or an oligomeric or glycosidically-bound derivative thereof

CYCLIC DIHYDROXY COMPOUNDS

PROCESS FOR THE PREPARATION OF OPTICALLY ACTIVE INTERMEDIATES FOR INSECTICIDAL COMPOUNDS

De-racemisation

Process for the preparation of prostaglandin precursors

MICROBIOLOGICAL OXIDATION PROCESSES

Isolation and activities of lignin depolymerases

Isolation and activities of lignin depolymerases.

A lignin depolymerase is effective for depolymerizing kraft lignin and delignifying kraft pulp. The lignin depolymerases can have pl values of about 4.9 and 5.8.

Isolation and activities of lignin depolymerases

FOR AN ENZYME TO THE CATALYSIS THE LIGNANBIOSYNTHESE CODING GENE AND USE OF IT

MODIFIED CYTOCHROM P450-MONOOXYGENASEN

PRODUCTION OF OPTICALLY ACTIVE ALCOHOLS WITH COMPLETE CELL CATALYSTS

SUCCINATUND MALONATSALZ OF TRANS-4 (1R, 3S) - 6CHLOR-3-PHENYLINDAN-1-YL) - 1,2,2TRIMETHYLPIPERAZIN AND the USE AS MEDICINE

MICROORGANISMS, THE ABILITY POSSESSIONS INDOL TO DEGRADIEREN AND EXCERPTS FROM IT, THE ONE INDOLOXIDASE ACTIVITY CREDIT

NEW CYTOCHROM P450-MONOOXYGENASEN AND THEIR USE FOR THE OXIDATION OF ORGANIC COMPOUNDS

MICRO-BIOLOGICAL PROCEDURE FOR the PRODUCTION OF 6-HYDROXYPYRAZINCARBONSÄURE

MIKROBI PRODUCTION OF PYROCATECHOLS.

PRODUCTION OF CELLS.

PROCEDURE FOR the PRODUCTION OF 4-HYDROXYZIMTALKOHOLEN

ELEKTRONENDONORSYSTEM FOR ENZYMES AND ITS APPLICATION DURING THE BIOCHEMICAL CONVERSION OF SUBSTRATES

PROCEDURE FOR THE PRODUCTION OF BUTANONE DERIVATIVES

PROCEDURE FOR THE PRODUCTION OF CONNECTIONS WITH HIGH MOLECULAR WEIGHT OF PHENOLIC CONNECTIONS ETC. AND APPLICATIONS THE SAME

Method comprising the indirect electrochemical regeneration of nad(p)h

Metabolically engineered cells for the production of pinosylvin

Metabolically engineered cells for the production of resveratrol or an oligomeric or glycosidically-bound derivative thereof

Production of stilbenoids

A method for the production of a stilbenoid, such as resveratrol or pinosylvin, by fermenting plant material such as grape must using a yeast having a metabolic pathway producing said stilbenoid, separating a solids waste material from said fermentation and extracting said stilbenoid.

METHOD FOR THE PRODUCTION OF RESVERATROL IN CELL CULTURES

Method of modulating metabolite biosynthesis in recombinant cells

MICROBIAL PRODUCTION OF CIS-DIHYDRODIOL AND PHENOL DERIVATIVES OF BENZOCYCLOBUTENE

Conversion of indene to (1s)-amino-(2r)-indanol free of any stereoisomer, by combination of dioxygenase bioconversion and chemical steps

PROCESS FOR PRODUCING 4-VINYLGUAIACOL BY BIODECARBOXYLATION OF FERULIC ACID

... 4-vinylguaiacol is produced using recombinant E.coli containing a decarboxylase gene from Bacillus pumilis in an aqueous fermentation broth and in an immobilized whole cell system. The 4-vinylguaiacol is extracted and recovered from an organic hydrocarbon solvent, preferably n-octane, whereby the product can readily be separated.

PROCESS FOR THE PREPARATION OF PHENOLIC CARBOXYLIC ACID DERIVATIVES BY ENZYMATIC CATALYSIS

The present invention relates to an improved process for the preparation of phenolic carboxylic acid derivatives catalysed by biocatalytic esterification, transesterification or amidation of a corresponding lower alkyl ester. Biocatalysis is performed in the presence of suitable enzymes, e.g. hydrolases, especially esterases, amidases, lipases and proteases.

METHOD OF MODULATING METABOLITE BIOSYNTHESIS IN RECOMBINANT CELLS

... ▓▓▓A method for modulating in a cell the expression of one or more genes involved ▓in the biosynthesis of a metabolite or a precursor for this metabolite by ▓inserting into a cell a nucleotide sequence coding for a transcription factor ▓comprising an AP2 DNA-binding domain, and/or by modifying the expression of a ▓nucleotide sequence coding for such a transcription factor already present in ▓the cell is provided. The method is useful for enhancing the biosynthesis of ▓secondary metabolites in plants such as alkaloids including terpenoid indole ▓alkaloids. Also provided is a method for enhancing stress tolerance in plants ▓by the use of such transcription factors.▓ ...

METHODS FOR THE PREPARATION OF OPTICALLY ACTIVE EPOXIDES AND VICINAL DIOLS FROM STYRENE EPOXIDES USING ENANTIOSELECTIVE EPOXIDE HYDROLASES DERIVED FROM YEASTS

The invention provides yeast strains, and polypeptides encoded by genes of such yeast strains, that have enantiospecific styrene epoxide hydrolase activity. The invention also features nucleic acid molecules encoding such polypeptides, vectors containing such nucleic acid molecules, and cells containing such vectors. Also embraced by the invention are methods for obtaining optically active styrene vicinal diols and optically active styrene epoxides.

METHOD FOR THE OXIDATION OF AROMATIC COMPOUNDS

The invention relates to a method for the biocatalytic oxidation of aromatic compounds, using recombinant micro-organisms expressing xylene monooxygenase, in a two-phase reaction medium.

AN ENZYME FOR CATALYZING BIOSYNTHESIS OF PIPERITOL AND/OR SESAMIN

... ²²²The present invention provides an enzyme that catalyzes the reaction producing ²piperitol from pinoresinol, and a reaction producing sesamin from piperitol. ²The invention also provides a gene that encodes such enzyme. Further, the ²invention provides a vector and transformant including a gene encoding the ²enzyme, and a producing method of the protein using the transformant.² ...

ENZYME CAPABLE OF REDUCING ACETOPHENONE AND OTHER CARBONYL COMPOUNDS AND PROCESSES THEREFOR

ELECTRON DONOR SYSTEM FOR ENZYMES AND ITS USE FOR THE BIOCHEMICAL CONVERSION OF SUBSTRATES

The invention relates to a novel electron donor system for enzymes with redox properties and to its use in enzyme-catalyzed oxidation reactions, such as the production of .omega. hydroxylated fatty acids. The invention further relates to an improved analysis method for fatty acid monoxygenases and to bioreactors and test kits wherein the inventive electron donor system is advantageously used.

MICROBIOLOGICAL OXIDATIONS

PROCESS FOR THE PREPARATION OF PROSTAGLANDIN PRECURSORS

A propargylic alcohol, enriched in the (R)-enantiomer, has formula (I) wherein R is C1-4 alkoxy, halogen, or C1-4 alkyl optionally substituted by OH or halogen. This may be prepared by the steps of: (a) enantioselective (R)- esterification of the racemic alcohol using an acyl donor and a first enzyme; (b) removal of the unreacted (S)-alcohol; and (c) enantioselective hydrolysis of the (R)-ester, using a second enzyme. The alcohol can be converted to a prostaglandin by known means.

METHOD COMPRISING THE INDIRECT ELECTROCHEMICAL REGENERATION OF NAD(P)H

MICROORGANISMS AND METHODS FOR THE FERMENTATION OF CANNABINOIDS

Disclosed herein are microorganism and methods that can be used for the synthesis of cannabigerolic acid (CBGA) and cannabinoids. The methods disclosed can be used to produce CBGA, ?9-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), cannabichromenic acid (CBCA), ?9 -tetrahydrocannabinol (THC), cannabidiol (CBD), cannabichromene (CBC). Enzymes useful for the synthesis of CBGA and cannabinoids, include but are not limited to acyl activating enzyme (AAE1), polyketide synthase (PKS), olivetolic acid cyclase (OAC), prenyltransferase (PT), THCA synthase (THCAS), CBDA synthase (CBDAS), CBCA synthase (CBCAS), HMG-Co reductase (HMG1), and/or farnesyl pyrophosphate synthetase (ERG20). The microorganisms can also have one or more genes disrupted, such as gene that that controls beta oxidation of long chain fatty acids.

BIOPRODUCTION OF AROMATIC CHEMICALS FROM LIGNIN-DERIVED COMPOUNDS

The teachings provided herein are generally directed to a method of converting lignin-derived compounds to valuable aromatic chemicals using an enzymatic, bioconversion process. The teachings provide a selection of (i) host cells that are tolerant to the toxic compounds present in lignin fractions; (ii) polypeptides that can be used as enzymes in the bioconversion of the lignin fractions to the aromatic chemical products; (iii) polynucleotides that can be used to transform the host cells to express the selection of polypeptides as enzymes in the bioconversion of the lignin fractions; and (iv) the transformants that express the enzymes.

MODIFICATION OF LIGNIN SYNTHESIS IN PLANTS

... 2118793 9305160 PCTABS00020 The biosynthesis of lignin in plants is regulated by insertion into the plant genome by altering the plant's ability to synthesize the enzyme O-methyl-transferase, an enzyme involved in the lignin biosynthetic pathway. Production of O-methyl-transferase may be enhanced by insertion into the plant genome by transformation of one or more additional copies of the O-methyl-transferase gene or production may be inhibited by insertion of a gene encoding anti-sense mRNA directed against the mRNA encoded by the endogenous O-methyl-transferase gene.

Enhance Indole Biosynthesis

RECOMBINANT PINORESINOL/LARICIRESINOL REDUCTASE, RECOMBINANT DIRIGENT PROTEIN, AND METHODS OF USE

Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated from Forsythia intermedia, Thuja plicata and Tsuga heterophylla, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases from these species. Accordingly, isolated DNA sequences are provided which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith (e.g., antisense dirigent protein or pinoresinol/lariciresinol reductase RNA or fragments of complementary dirigent protein or pinoresinol/lariciresinol reductase DNA which are useful as polymerase chain reaction primers or as probes for genes encoding dirigent proteins or pinoresinol/lariciresinol ...

INTERMEDIATE PRODUCT FOR PREPARATION OF LIGNIN POLYMERS AND USE THEREOF FOR PRODUCTION OF WOOD MATERIALS

The invention relates to an intermediate product for the preparation of polymerizates from lignin derivatives obtained from the cellulose industry. The intermediate product is produced by treating the lignin derivatives with enzymes capable of oxidizing phenol in the presence of oxidizing agents. According to the invention the lignin derivatives are subjected to the enzymatic treatment for more than 3 hours in the presence of air, or they are subjected to an enzymatic treatment for more than 10 min, whereby air or oxygen is introduced. The lignin polymers obtained from the polymerization of lignin derivatives performed in the presence of active intermediate products are suited for use in the production of binding agents for wood materials.

MEDICINAL USES OF PHENYLALKANOLS AND DERIVATIVES

A compound of formula (I), a pharmaceutically acceptable derivative thereof, wherein Ph is a phenyl radical R1 is H, OH, OC1-4alkyl, NO2; R2 is OH, OC1-4alkyl, OC=OC1-4alkyl or OC=OPh where the Ph can be optionally substituted by halogen, C1-3 alkyl or NO2; R1 and R2 along with the two carbon atoms of the phenyl ring to which they are attached can combine to form a 5 or 6 membered heterocyclic ring comprising 1 or 2 heteroatoms selected from O, S or N; R3 is an optionally substituted hydrocarbyl radical; R4 is H, CH3, OH or =O; when R4 is =O, then the carbon to which R4 is attached is not bonded to H; W is C(=O)-CH2, CH=CH-, CH2CO, CH(OH)-CH2, C(CH3)(OH)CH2, CH2CH(OH), CH2C(CH3)OH, CO, CHOH, C(CH3)(OH), CH2, CH2CH2; X is -CH-OH, C(CH3)OH, CH2, CH(CH3) or -C=O; Y is -CH-OH, C(CH3)OH, CH2, CH(CH3) or -C=O; provided that one of W, X or Y has an OH group.

Verfahren zur Reduktion von Epoxyketonen

Microbiological synthesis of methyl muconicacids

Microbiological synthesis. New prepn. e.g. of alpha, alpha'-dimethylmuconic acid No cardia (where R1, R2, R3 = H or CH3). Pref. strains of Nocardia are ATCC Nos. 19070 and 19071).

2, 3-dihydroxybenzoic acid opt methyl-substd - prodn by fermentation of methylbenzenes with nocardia microorganisms

Organic acids having the general formula: (in which R1, R2 and R3 are H or CH3) are produced by subjecting toluene opt. substd. with R1, R2 and R3 to fermentation by Nocardia corallina, salmonicolor or minima in presence of a nutrient liquid. The products can be used as chelating agents, metal de-activators, dye intermediates, etc.

Verfahren zur Herstellung eines Mycels

КОЛИЧЕСТВЕННОЕ ПРЕВРАЩЕНИЕ ИНДЕНА В (1S, 2R)-ОКСИД ИНДЕНА И (1S,2R)-ИНДАНДИОЛ ПУТЕМ СОЧЕТАНИЯ БИОЛОГИЧЕСКОГО ПРЕВРАЩЕНИЯ ПОД ДЕЙСТВИЕМ ГАЛОИДПЕРОКСИДАЗЫ И ХИМИЧЕСКИХ СТАДИЙ.

ЕЛЕКТРОФЕРМЕНТАТИВНИЙ СПОСІБ ОДЕРЖАННЯ 2,3-ДИГІДРОКСИФЕНІЛЬНИХ ПОХІДНИХ ТА СПОСІБ ЕЛЕКТРОХІМІЧНОЇ РЕГЕНЕРАЦІЇ NAD(P)H З NAD(P)+

Винахід належить до способів, що включають непряму електрохімічну регенерацію NAD(P)H при ферментативних перетвореннях субстратів, що каталізуються монооксигеназами. Об'єктом винаходу є спосіб електроферментативного одержання 2,3-дигідроксифенільних похідних при каталізі ферментом 2-гідроксибіфеніл-3-монооксигеназою при одночасному електрохімічному відновленні NAD(P)+, що утворився в результаті відновного розщеплення кисню.

METHOD FOR THE ELECTROENZYMATIC PRODUCTION OF 2,3-DIHYDROXYPHENYL DERIVATIVES AND METHOD FOE ELECTROCHEMICAL REGENERATION OF NAD(P)H FROM NAD(P)+

SOLID FORMS OF MENAQUINOLS

ТВЕРДЫЕ ФОРМЫ МЕНАХИНОЛОВ

Описан способ получения стабильной кристаллической формы менахинола восстановлением менахинона. Пункты: 1 независимый 3 зависимых Фиг. 6 ...

Preparation method of pulegone derivative

Method for catalytic synthesis of R-2-bromo-1-aryl alcohol using carrot root whole cells

Β - glucosidase high yield and its application in the conversion of resveratrol

Method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis

New application of GDSL protein to preparation of lipase

The invention discloses new application of GDSL protein to preparation of lipase. The invention provides application of the GDSL protein to (1) preparation of lipases or (2) degradation of a substrate of the lipase. The GDSL protein is the following protein in (a) or (b): (a) the protein comprises the amino acid sequence shown as the sequence 2; and (b) the protein is formed in the way that the amino acid sequence shown as the sequence 2 is subjected to substitution and/or deletion and/or addition of one or more amino acid residues, has lipase activity and is derived from the sequence 2. The GDSL protein has lipase activity. The application of the GDSL protein as the lipase has the advantages of higher reaction temperature, high heat stability, action under the alkaline condition, high pH stability, effects on the substrates of C2-C12 and high enzyme activity. The invention opens up a new way for the process flow of the lipase and has a great economic value.

Cresol trimer compounds as well as preparation method and application thereof

The invention relates to cresol trimer compounds as well as a preparation method and application thereof. The compounds with novel structure are prepared from Penicillium expansum 091006 separated from a root sample of Excoecaria agallocha which is a mangrove plant. Proved by experiments, the compounds can be used as a cell proliferation inhibitor or antitumor agent.

Biological preparation method (1R,2S)-2-(3,4- difluorophenyl) cyclopropylamine D- mandelic acid salt (I)

Process for preparing high-purity resveratrol from giant knotweed

Meyerozyma guilliermondii 3-J15 and application thereof

Use Of "Polyketide Synthases" Type Iii (Pks Iii) Recombinant Marine Brown Algae

A METHOD FOR THE BIOHEMICAL ISOLATION OF l-MENTHOL

USE OF “POLYKETIDE SYNTHASES” OF TYPE III (PKS III) RECOMBINING MARINE BROWN ALGAS

La présente invention concerne une méthode de production de composés polyphénoliques, i.e. le phloroglucinol ou l'un de ses dérivés, par une polyketide synthase de type III (PKSIII) d'algue brune marine. L'invention concerne également des acides nucléiques recombinants codant pour une polyketide synthase de type III (PKSIII) de l'algue brune Ectocarpus siliculosus (E. siliculosus), des vecteurs recombinants comprenant ces acides nucléiques, ainsi que des cellules hôtes comprenant ces vecteurs. Enfin, l'invention concerne une méthode de préparation de divers composés à l'aide des composés polyphénoliques produits selon la méthode précitée.

Synthesis of propargyl alcohol derivatives involves stereoselective acylation of a racemic propargyl alcohol in the presence of a lipase, solvent and acylating agent, via a Mitsunobu reaction on the mixture of alcohol and acetate

L'invention se rapporte à un nouveau procédé de synthèse énantiosélective de composés organiques de formule générale (I) : Le dit procédé étant industrialisable et permettant d'obtenir à partir d'un mélange d'alcools racémiques de formule (II), les alcools chiraux correspondants sous une forme énantiomériquement pure, avec un rendement chimique élevé.

CORYNEFORM BACTERIUM TRANSFORMANT AND METHOD FOR PRODUCING PHENOL USING SAME

Synthesis of bis-eugenol from eugenol with suspension cell of Kalopanax pictus Nakai

PURPOSE: A method for synthesizing bis-eugenol through bioconversion is provided to ensure antibacterial, skin whitening, and antioxidation effects. CONSTITUTION: A method for synthesizing bis-eugenol comprises: a step of preparing a culture liquid in which Kalopanax pictus is cultured; and a step of adding eugenol to the culture liquid and culturing. The method further comprises a step of adding EtOAc to the culture liquid and stirring; and a step of evaporating the EtOAc. The preparation of the culture liquid comprises a step of inducing callus from Kalopanax pictus using a solid medium; and a step of culturing the callus in a liquid medium at 15-30°C. COPYRIGHT KIPO 2012 ...

PROCESS FOR PREPARING HYDROQUINONE AND BACTERIA OF THE [...][...][...] OB3B ADAPTED FOR GROWTH IN METHANOL

Oxidoreductase and processes for producing 3-(p-hydroxyphenyl)-2-propenol derivatives and other compounds using the same

A novel enzyme which is obtained from a microorganism of a strain belonging to the genus Pseudomonas fluorescens and catalyzes the oxidation of substrates such as p-hydroxytoluene derivatives and p-alkylphenol derivatives; and processes for producing 3-(p-hydroxyphenyl)-2-propenol derivatives, p-hydroxybenzaldehyde derivatives, p-alkylphenol derivatives, optically active S(-)-1-(4-hydroxyphenyl) alcohol derivatives, etc., from those substrates using the enzyme.

REMOVING POLYPHENOL CONTAMINANTS FROM FEEDSTOCK-BASED POLYPHENOLS

Provided is a method of producing a mixture of pure feedstock-based native polyphenols from a feedstock. Contaminant polyphenols are first removed from an enzyme solution for converting feedstock to a product to produce a polyphenol reduced enzyme solution. The polyphenol reduced enzyme solution is combined with the feedstock and the feedstock is converted to a product and by-product. Heretofore, there has been no process available to reduce or remove the contaminant phenols introduced to the feedstock by commercial enzyme solutions. This method allows for the removal of contaminant phenols prior to introduction to the processing stream and subsequent harvesting of pure feedstock6based native polyphenols. The pure feedstock-based polyphenols are removed from the product or by-product to produce a pure mixture of feedstock-based polyphenols.

PRODUCTION OF ANTIMICROBIALS FROM VEGETABLE WASTE

The present invention relates to a process for preparing an extract with antimicrobial activity from by-products or waste products of the fruit and vegetable industry. The process comprises a step of mixing the by-products or waste products with at least one microorganism, preferably a bacterium of the genus Lactobacillus, a fermentation step and a subsequent extraction step.

METHOD FOR GENERATING 3,5-DIHYDROXY-4-METHOXYBENZYL ALCOHOL FROM PLANKTON

... [Problem] The purpose of the present invention is to provide a method for collecting seawater that contains plankton and generating DHMBA (dba), which is an antioxidant, from the plankton contained in the seawater. [Solution] The present invention is characterized in that: collected seawater containing plankton is filtered using a filter, the cell contents are removed from the plankton remaining on the filter, the removed cell contents are subsequently heated, and 3,5-dihydroxy-4-methoxybenzyl alcohol is produced from the heated product; and the plankton are assumed to be diatoms.

SESQUITERPENE SYNTHASES AND METHODS OF USE

The invention relates to sesquiterpene synthases and methods of their production and use. In one embodiment, the invention provides nucleic acids comprising a nucleotide sequence as described herein that encodes for at least one sesquiterpene synthases. In a further embodiment, the invention also provides for sesquiterpene synthases and methods of making and using these enzymes. For example, sesquiterpene synthases of the invention may be used to convert famesyl-pyrophosphate to various oxygenated and aliphatic sesquiterpenes including valencene, bicyclo-germacrene, cubebol and delta- cadinene.

METHODS OF OBTAINING OPTICALLY ACTIVE EPOXIDES AND VICINAL DIOLS FROM STYRENE OXIDES

The invention provides yeast strains, and polypeptides encoded by genes of such yeast strains, that have enantiospecific styrene epoxide hydrolase activity. The invention also features nucleic acid molecules encoding such polypeptides, vectors containing such nucleic acid molecules, and cells containing such vectors. Also embraced by the invention are methods for obtaining optically active styrene vicinal diols and optically active styrene epoxides.

DITERPENE CYCLASE AND METHODS OF USE

An enzyme having diterpene cyclase activity has been purified from P. elisabethae using a series of chromatogrqaphy steps. The purified enzyme has an apparent molecular weight of about 47 kilodaltons and an isoelectric point of about 5.1. The purified enzyme catalyzed the cyclization of geranyl geranyl diphosphate to elisabethatriene.

Microorganisms and methods for the biosynthesis of aromatics, 2,4-pentadienoate and 1,3-butadiene

The invention provides non-naturally occurring microbial organisms having a toluene, benzene, p-toluate, terephthalate, (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate, (2-hydroxy-4-oxobutoxy)phosphonate, benzoate, styrene, 2,4-pentadienoate, 3-butene-1ol or 1,3-butadiene pathway. The invention additionally provides methods of using such organisms to produce toluene, benzene, p-toluate, terephthalate, (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate, (2-hydroxy-4-oxobutoxy)phosphonate, benzoate, styrene, 2,4-pentadienoate, 3-butene-1ol or 1,3-butadiene.

Process for the stereoselective enzymatic reduction of keto compounds

In a process for the stereoselective, in particular enantioselective enzymatic reduction of keto compounds to the corresponding chiral hydroxy compounds, wherein the keto compounds are reduced with an enantioselective, NADH-specific oxidoreductase, a polypeptide is used for reducing the keto compounds, which polypeptide exhibits an R-ADH-signature H-[P; A]-[I; A; Q; V; L]-[G; K]-R at positions 204-208 and the following further structural features in their entirety: (i) an N-terminal Rossmann-Fold GxxxGxG, (ii) an NAG-motif at position 87, (iii) a catalytic triad consisting of S 139, Y 152 and K 156, (iv) a negatively charged amino acid moiety at position 37, (v) two C-terminal motifs in the dimerization domain [A; S]-S-F and [V; I]-DG-[G; A]-Y-[T; C; L]-[A; T; S]-[Q; V; R; L; P], (vi) Val or Leu at position 159 (4 positions downstream of K 156), (vii) Asn at position 178, and (viii) a proline moiety at position 188.

Buffer composition for catalyzing the preparation of calcitriol or calcifediol and method for preparing calcitriol or calcifediol using same

The present invention relates to a buffer composition for promoting production of calcitriol or calcifediol, and a method for producing calcitriol or calcifediol using the same. More particularly, the present invention relates to a buffer composition for promoting production of calcitriol or calcifediol comprising a metallic compound, an organic solvent, cyclodextrin, tris(hydroxymethyl)aminomethane, sodium succinate, sodium chloride, magnesium chloride, and water, and a method for producing calcitriol or calcifediol using the same. In the method for producing calcitiriol or calcifediol, the production yield of calcitriol or calcifediol is high, and the bioconversion is carried out in an enzyme reaction system instead of in a microorganism culture system. Thus, it is not required to maintain a sterile state. Also, the separation/purification following the completion of a biocatalytic reaction can be carried out in a cleaner state than the microorganism culture method. Accordingly, there is an advantage in that a cost required for separation is low and the quality is improved. Furthermore, the buffer composition for promoting production of calcitriol or calcifediol can provide a high productivity of calcitriol or calcifediol.

Production Of Pure Lignin From Lignocellulosic Biomass

The present invention is directed to a process of producing substantially pure lignin from lignocellulosic biomass, which comprises: pre-treating a lignocellulosic feedstock to produce a reactive lignin-carbohydrate mixture; biologically-reacting the carbohydrates in the mixture, separating remaining solids from the liquid fermentation products, and drying the resulting solids to yield a substantially pure lignin product. Optionally, the lignin product may be washed and subjected to a second hydrolysis step. Optionally, the lignin product may be further processed by hydrotreating and/or pyrolysis in order to yield desirable products such as fuel additives.

Method for producing microbial fermentation product

Disclosed is a method for producing 1-(2-hydroxyethyl) 2,5,5,8a-tetramethyldecahydronaphthalene-2-ol represented by formula (2), wherein microbial conversion is carried out using a compound(s) represented by formula (1a) and/or (1b) as a substrate, the resulting culture product, in which microorganisms obtained by the microbial conversion are contained, and a solvent having an SP value within the range of 7.5 to 9.0 [(cal/cm 3 ) 1/2 ] are mixed together, and subsequently the aqueous phase is removed therefrom.

KETOREDUCTASE POLYPEPTIDES FOR THE REDUCTION OF ACETOPHENONES

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. 1Lactobacillus. A method for stereoselectively reducing a 2′ ,6′-substituted acetophenone substrate , optionally substituted at one or more positions selected from the group consisting of 3′ , 4′ , and 5′ , to the corresponding substituted (S)-1-phenethanol , which comprises contacting the substrate with an engineered ketoreductase polypeptide under reaction conditions suitable for stereoselectively reducing or converting the substrate to the corresponding substituted (S)-1-phenethanol product , wherein the engineered ketoreductase polypeptide is derived from a wild-type ketoreductase and is capable of stereoselectively reducing acetophenone to (S)-1-phenethanol with a percent stereomeric excess of at least about 90%.2. The method of claim 1 , wherein the substrate is 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone and the corresponding substituted (S)-1-phenethanol product is (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl)ethanol.3. The method of claim 1 , wherein the (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl)ethanol is formed in greater than 99% stereomeric excess.4. The method of claim 1 , wherein at least about 95% of the substrate is reduced to the product in less than 24 hours when the method is conducted with at least 200 g/L of substrate and with less than 2 g/L of the polypeptide.5. The method of claim 1 , wherein the method is carried out with whole cells that express the engineered ketoreductase polypeptide claim 1 , or an extract or lysate of such cells.6. The method of claim 1 , wherein the engineered ketoreductase polypeptide is isolated and/or ...

CORYNEFORM BACTERIUM TRANSFORMANT AND PROCESS FOR PRODUCING PHENOL USING THE SAME

A phenol-producing transformant constructed by transferring a gene which encodes an enzyme having tyrosine phenol-lyase activity into as a host can efficiently produce phenol from a saccharide. Specifically, preferred is a process which comprises a step of reacting the transformant in a reaction mixture containing a saccharide under reducing conditions, and a step of collecting phenol from the reaction mixture. 1. A phenol-producing transformant constructed by transferring a gene which encodes an enzyme having tyrosine phenol-lyase activity into a coryneform bacterium as a host.2Pantoea agglomeransCitrobacter braakiiDesulfitobacterium hafnienseChloroflexus aurantiacusNostoc punctiformeTreponema denticola.. The transformant of claim 1 , wherein the gene which encodes an enzyme having tyrosine phenol-lyase activity is a gene derived from claim 1 , a gene derived from claim 1 , a gene derived from claim 1 , a gene derived from claim 1 , a gene derived from claim 1 , or a gene derived from3. The transformant of claim 1 , wherein the gene which encodes an enzyme having tyrosine phenol-lyase activity is the DNA of the following (a) or (b).(a) a DNA consisting of the base sequence of SEQ ID NO: 36, a DNA consisting of the base sequence of SEQ ID NO: 39, a DNA consisting of the base sequence of SEQ ID NO: 42, a DNA consisting of the base sequence of SEQ ID NO: 45, a DNA consisting of the base sequence of SEQ ID NO: 48, or a DNA consisting of the base sequence of SEQ ID NO:(b) a DNA which hybridizes to a DNA consisting of a complementary base sequence of any of the DNAs of (a) under stringent conditions and which encodes a polypeptide having tyrosine phenol-lyase activity4. The transformant of claim 1 , wherein the following gene (c) and/or gene (d) on the chromosome of the coryneform bacterium as the host has a disruption or deletion.(c) a gene which encodes an enzyme having prephenate dehydratase activity(d) a gene which encodes an enzyme having phenol 2-monooxygenase ...

ENANTIOSELECTIVE REDUCTION

The invention relates to a process for the preparation of a chiral compound, comprising enantioselectively reducing a carbon-carbon double bond of an α,β-unsaturated compound in a mixture comprising both an E isomer and a Z isomer of the α,β-unsaturated compound, wherein both E isomer and Z isomer are converted in the presence of a hydrogenation catalyst. 1. Process for the preparation of a chiral compound , comprising enantioselectively reducing a carbon-carbon double bond of an α ,β-unsaturated compound in a mixture comprising both an E isomer and a Z isomer of the α ,β-unsaturated compound , wherein both E isomer and Z isomer are converted in the presence of a catalyst , and wherein the reduction is carried out under isomerising conditions.2. Process according claim 1 , wherein the molar ratio Z isomer to E isomer in the mixture at the start of the process is in the range of 5:95 to 95:5 claim 1 , in particular 10:90 to 90:10 claim 1 , more in particular 20:80 to 80:20.3. Process according to claim 1 , wherein the chiral compound is formed with an enantiomeric excess of at least 50% claim 1 , in particular of at least 80% claim 1 , more in particular of at least 90%.4. Process according to claim 1 , wherein the catalyst is a biocatalyst claim 1 , in particular an enzyme which enzyme may be present in an organism or isolated from an organism claim 1 , and which enzyme preferably is an oxidoreductase claim 1 , more preferably an ene reductase.5. Process according to claim 4 , wherein the biocatalyst is an enzyme and reduction is carried out in the presence of a cofactor regeneration system for the enzyme.6. Process according to claim 4 , wherein the enzyme is a substrate unspecific enzyme.7. Process according to claim 4 , wherein the enzyme is a substrate specific enzyme.8. Process according to claim 4 , wherein the enzyme is selected from the group of ene reductases HYE1 claim 4 , HYE2 claim 4 , P1 and LTB4DH.9. Process according to claim 1 , wherein the ...

PRODUCTION OF INDUSTRIALLY RELEVANT COMPOUNDS IN PROKARYOTIC ORGANISMS

Disclosed herein are methods for producing compounds (such as 3,4-dihydroxybenzoate, catechol, cis,cis-muconate, or β-carboxy-cis,cis-muconic acid) utilizing biosynthetic pathways in prokaryotic organisms expressing one or more heterologous genes. In some embodiments, the method includes expressing a heterologous asbF gene (for example, a gene having dehydroshikimate dehydratase activity) in a prokaryotic cell under conditions sufficient to produce the one or more compounds and purifying the compound. In additional embodiments, the method further includes expressing one or more of a heterologous 3,4-DHB decarboxylase gene, a heterologous catechol 1,2-dioxygenase gene, and a heterologous 3,4-DHB dioxygenase gene in the prokaryotic cell and purifying the compound. 1. A method for producing a compound utilizing dehydroshikimate as a precursor , wherein the compound is selected from 3 ,4-dihydroxybenzoate (3 ,4-DHB) , catechol , cis ,cis-muconate , and β-carboxy-cis ,cis-muconic acid , comprising:expressing a heterologous gene encoding a protein having dehydroshikimate dehydratase (DHSase) activity in a prokaryotic cell that is a phototroph under conditions sufficient to produce the compound; andpurifying the compound.2. The method of claim 1 , wherein the phototroph is a cyanobacterium.3SynechocystisSynechocystisSynechococcusSpirulinaAnabaena variabilis.. The method of claim 2 , wherein the cyanobacterium is PCC6803 claim 2 , PCC9714 claim 2 , sp. claim 2 , sp. claim 2 , or4Bacillus thuringiensis, Bacillus cereusBacillus anthracis.. The method of claim 1 , wherein the heterologous gene encoding a protein having dehydroshikimate dehydratase (DHSase) activity comprises a gene from claim 1 , or5. The method of claim 1 , wherein the heterologous gene encoding a protein having dehydroshikimate dehydratase (DHSase) activity comprises the nucleic acid sequence set forth as SEQ ID NO: 1 or SEQ ID NO: 3.6. The method of claim 1 , wherein the heterologous gene encoding a protein ...

CORYNEFORM BACTERIUM TRANSFORMANT AND PROCESS FOR PRODUCING PHENOL USING THE SAME

Provided is a phenol-producing transformant constructed by transferring a gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity into as a host. Also provided is a process for producing phenol, which comprises a step of allowing the transformant to react in a reaction mixture containing 4-hydroxybenzoate or a salt thereof under reducing conditions, and a step of collecting phenol from the reaction mixture. 1Corynebacterium glutamicum. A phenol-producing transformant constructed by transferring a gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity into as a host.2Bacillus subtilis, Bacillus atrophaeus, Bacillus subtilisCitrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae, Enterobacter hormaechei, Enterobacter sakazakii, Escherichia coli, Escherichia fergusonii, Paenibacillus polymyxaPantoea ananatis.. The transformant of claim 1 , wherein the gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity is a gene derived from subsp. spizizenii claim 1 , claim 1 , or3. The transformant of claim 1 , wherein the gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity is the DNA of the following (a) or (b).(a) a DNA consisting of the base sequence of SEQ ID NO: 16, SEQ ID NO: 23, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 35, SEQ ID NO: 38, SEQ ID NO: 41, SEQ ID NO: 44, NO: 47, SEQ ID NO: 50, or SEQ ID NO: 53(b) a DNA which hybridizes to a DNA consisting of a complementary base sequence of any of the DNAs of (a) under stringent conditions and which encodes a polypeptide having 4-hydroxybenzoate decarboxylase activity.4Corynebacterium glutamicumCorynebacterium glutamicum. The transformant of claim 1 , wherein the as the host is a strain of in which a gene which encodes an enzyme having phenol 2-monooxygenase activity on the chromosome is disrupted or deleted.5Corynebacterium glutamicumCorynebacterium glutamicum. The transformant of claim 1 , wherein the as the host ...

Coryneform bacterium transformant and process for producing phenol using the same

Provided is a phenol-producing transformant constructed by transferring a gene which encodes an enzyme having tyrosine phenol-lyase activity into a coryneform bacterium as a host. Also provided is a process for producing phenol, which comprises a step of allowing the transformant to react in a reaction mixture containing tyrosine, a salt thereof, or an ester thereof under reducing conditions, and a step of collecting phenol from the reaction mixture.

CORYNEFORM BACTERIUM TRANSFORMANT AND PROCESS FOR PRODUCING PHENOL USING THE SAME

Provided is a phenol-producing transformant constructed by transferring a gene which encodes an enzyme having chorismate-pyruvate lyase activity and a gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity into a coryneform bacterium as a host. Also provided is a process for producing phenol, which comprises a step of allowing the transformant to react in a reaction mixture containing a saccharide under reducing conditions, and a step of collecting phenol from the reaction mixture. 1. A phenol-producing transformant constructed by transferring a gene which encodes an enzyme having chorismate-pyruvate lyase activity and a gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity into a coryneform bacterium as a host.2Escherichia coliPseudomonas putidaAcinetobacter baumanniiAzotobacter vinelandiiChromohalobacter salexigensCitrobacterCitrobacter koseriCitrobacter youngaeEnterobacter cloacaeMarinobacter aquaeoleiMarinomonas mediterraneaPantoea ananatisPseudoalteromonas haloplanktisRalstonia eutrophaShewanella putrefaciensThiobacillus denitrificans.. The transformant of claim 1 , wherein the gene which encodes an enzyme having chorismate-pyruvate lyase activity is a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from members of the genus claim 1 , such as and ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; a gene derived from ; or a gene derived from3Bacillus subtilisBacillus atrophaeusBacillus subtilisspizizeniiCitrobacter koseriEnterobacter aerogenesEnterobacter cloacaeEnterobacter hormaecheiEnterobacter sakazakiiEscherichia coliEscherichia fergusoniiPaenibacillus polymyxaPantoea ananatis.. The transformant of claim 1 , wherein the gene which encodes an enzyme having 4-hydroxybenzoate decarboxylase activity is a gene derived from claim 1 , a gene derived from claim 1 , ...

Method for Producing Probiotically Derived Compounds

A method for producing naturally derived beneficial compounds including dispersing a microbiological culture media including at least one live probiotic organism, and at least one nutraceutical and/or at least one nutritive agent in distilled water to form a broth, incubating the broth at a predetermined temperature for a select period of time to induce probiotic activity; halting the probiotic activity, and separating the desired compound from the broth. 1. A process for naturally deriving a beneficial compound comprising:preparing a microbiological culture comprising at least one live probiotic organism and at least one nutritive agent or at least one nutraceutical agent;incubating the microbiological culture to initiate probiotic activity;halting the probiotic activity;harvesting a waste byproduct of the probiotic activity; andseparating the beneficial compound from the waste byproduct.2LactobacillusBifidobacteriumEnterococcusStreptococcus thermophilus. The process of wherein the at least one live probiotic organism is selected from the group consisting of species claim 1 , species claim 1 , species claim 1 , claim 1 , and combinations thereof.3. The process of wherein the microbiological culture is incubated at a temperature of from about 35° C. to about 40° C.4. The process of wherein the microbiological culture is incubated for a period of from about 24 hours to about 240 hours.5. The process of wherein the probiotic activity is halted by adding organic ethanol.6. A process for naturally deriving at least one B vitamin coenzyme comprising:{'i': Lactobacillus', 'Bifidobacterium', 'Enterococcus', 'Streptococcus thermophilus, 'preparing a microbiological culture comprising at least one live probiotic organism selected from the group consisting of species, species, species, , and combinations thereof, at least one nutritive agent, and at least one source of a B vitamin;'}incubating the microbiological culture to initiate probiotic activity;halting the probiotic ...

Metabolically Engineered Cells For The Production of Pinosylvin

A genetically engineered micro-organism having an operative metabolic pathway producing cinnamoyl-CoA and producing pinosylvin therefrom by the action of a stilbene synthase is used for pinosylvin production. Said cinnamic acid may be formed from L-phenylalanine by a L-phenylalanine ammonia lyase (PAL) which is one accepting phenylalanine as a substrate and producing cinammic acid therefrom, preferably such that if the PAL also accepts tyrosine as a substrate and forms coumaric acid therefrom, the ratio Km(phenylalanine)/Km(tyrosine) for said PAL is less than 1:1 and if said micro-organism produces a cinammate-4-hydroxylase enzyme (C4H), the ratio K(PAL)/K(C4H) is at least 2:1. 163-. (canceled)64. A micro-organism having an operative metabolic pathway producing cinnamoyl-CoA and producing pinosylvin therefrom by the action of a stilbene synthase , wherein said cinnamic acid is formed from L-phenylalanine by a L-phenylalanine ammonia lyase (PAL) which is one accepting phenylalanine as a substrate and producing cinammic acid therefrom , such that if the PAL also accepts tyrosine as a substrate and forms coumaric acid therefrom , the ratio Km(phenylalanine)/Km(tyrosine) for said PAL is less than 1:1.65. A micro-organism as claimed in claim 64 , wherein if said micro-organism produces a cinammate-4-hydroxylase enzyme (C4H) claim 64 , the ratio K(PAL)/K(C4H) is at least 2:1.66. A micro-organism as claimed in claim 65 , wherein cinnamoyl-CoA is formed in a reaction catalysed by an enzyme in which ATP and CoA are substrates and ADP is a product.67. A micro-organism as claimed in claim 66 , wherein cinnamoyl-CoA is formed in a reaction catalysed by a 4-coumarate-CoA ligase or cinnamate-CoA ligase.68. A micro-organism as claimed in claim 67 , wherein said 4-coumarate-CoA ligase or cinnamate-CoA ligase is expressed in said micro-organism from nucleic acid coding for said enzyme which is not native to the micro-organism. ...

Microorganisms And Methods For Producing Substituted Phenols

The present invention is concerned with methods for producing vanillin, feroyl-CoA, ferulic acid, coniferyl aldehyde and/or coniferyl alcohol. Also, the invention relates to microorganisms useful in such production method, and to the construction of such microorganisms. 1. Method for producing a product selected from the group consisting of vanillin , feruloyl-CoA , ferulic acid , coniferyl aldehyde and coniferyl alcohol , comprising adding an educt selected from the group consisting of eugenol , coniferyl alcohol , coniferyl aldehyde and ferulic acid to a microorganism of order Actinomycetales , wherein the microorganism does not comprise a gene coding for a vanillin dehydrogenase.2. The method according to claim 1 , wherein the vanillin dehydrogenase has an amino acid sequence according to any of SEQ ID 1 claim 1 , 2 or 3.3. The method according to claim 1 , wherein the microorganism comprises a vanillin dehydrogenase gene inactivated by an insert or a deletion.4. The method according to claim 1 , wherein the microorganism comprises a gene selected from the group consisting of:{'i': Pseudomonas', 'Penicillium simplicissimium', 'Rhodococcus jostii, 'an eugenol hydroxylase gene of sp. DSMZ 7063, a vanillyl alcohol oxidase gene of , and an eugenol oxidase gene of RHA1,'}{'i': Pseudomonas', 'Rhodococcus opacus', 'Acinetobacter', 'Rhodococcus erythropolis', 'Lactobacillus plantarum', 'Gordonia polyisoprenivorans, 'sub': —', '—, 'a coniferyl alcohol dehydrogenase gene of sp. DSMZ 7063, an aromatic alcohol dehydrogenase gene (adhA; accession number AB213394) of TKN14, an aryl alcohol dehydrogenase gene of sp. ADP1, an aryl alcohol dehydrogenase gene of PR4, WCFS 1 (lp3054; accession number CAB69495) and Kd2 (adhA, accession number ZP09272609.1),'}{'i': Pseudomonas', 'Acinetobacter', 'Arabidopsis thaliana', 'Pseudomonas putida', 'Gluconobacter oxydans, 'sub': —', '—, 'a coniferyl aldehyde dehydrogenase of sp. DSMZ 7063, a benzaldehyd dehydrogenase of sp. ADP1, an aldehyd- ...

POLYPEPTIDES HAVING DEMETHYLATING ACTIVITY

The present invention relates to fungal enzymes, more in particular of polyphenoloxidases (PPOs), and is based on a newly discovered enzymatic activity of a class of PPOs, i.e. de-methylation of R-substituted mono- or di-methoxyphenolsuch as present in lignin, lignin derived compounds and/or in lignocellulosic biomass. This newly discovered enzymatic activity renders these enzymes highly suitable for a plethora of applications in industry. Provided herein are methods of demethylation, processes to increase the reactivity of lignin or lignin-comprising biomass, processes of conversion lignin or lignin comprising biomass to value added products, processes for degrading and/or modifying (hemi-)cellulose in a hemicellulose-comprising substrate, and expression vectors, host cells and liquids, pastes or solid formulations and compositions for use in the demethylation method of the invention. 2. A polyphenoloxisase according to that comprises or consists of an amino acid sequence that is at least 70% identical to the amino acid sequence of at least one of SEQ ID NO: 37 claim 1 , 41 and 45.3. A polyphenoloxisase according to or that is an enzyme that releases methanol from a R-substituted di-methoxyphenol represented by formula [1].4Myceliophthora.. A polyphenoloxisase according to any one of the preceding claims that is obtainable from a fungus claim 1 , preferably a5Myceliophthora thermophila C. A polyphenoloxisase according to any one of the preceding claims that is obtainable from 1.6. A polyphenoloxisase according to any one of the preceding claims claim 1 , wherein R is an organic moiety.7. A method of demethylation of an R-substituted mono- or di-methoxyphenol represented by formula [1] claim 1 , [2] claim 1 , or [3] claim 1 , comprising the step of contacting a substrate comprising said R-substituted mono- or di-methoxyphenol with a polyphenoloxidase of any one of the - claim 1 , wherein R can be any single atom or chemical moiety.8. A method according to claim 7 , ...

METHODS OF MAKING CAPSINOIDS BY BIOSYNTHETIC PROCESSES

Provided herein are methods of making capsinoids including providing a capsiate synthase in a mixture or cellular system, feeding 8-methyl-6-nonenoyl-CoA, 6E-8-methylnonenoic acid or 8-methylnonanoic acid into the mixture or cellular system, feeding vanillyl alcohol into the mixture or cellular system, and collecting capsinoids from the mixture or cellular system. 1. A method of producing a capsinoid , the method comprising:(a) expressing a capsiate synthase (CS) in a cellular system;(b) adding 8-methyl-6-nonenoyl-CoA and vanillyl alcohol to the cellular system; and(c) incubating the cellular system for a sufficient time to produce the capsinoid.2. A method of producing dihydrocapsiate , the method comprising:(a) expressing a capsiate synthase (CS) and an acyltransferase (ACS) in a cellular system;(b) adding 8-methylnonanoic acid and vanillyl alcohol to the cellular system; and(c) incubating the cellular system for a sufficient time to produce the dihydrocapsiate.3. A method of producing capsiate , the method comprising:(a) expressing a capsiate synthase (CS) and an acyltransferase (ACS) in a cellular system;(b) adding 6E-8-methylnonanoic acid and vanillyl alcohol to the cellular system; and(c) incubating the cellular system for a sufficient time to produce the capsiate.4. A method of producing a capsinoid , the method comprising:(a) expressing a capsiate synthase (CS) and an acyltransferase (ACS) in a cellular system;(b) adding a medium chain fatty acid and vanillyl alcohol to the cellular system; and(c) incubating the cellular system for a sufficient time to produce the capsinoid.5Capsicum. The method of any one of to , wherein the CS amino acid sequence is derived from a plant of the genus.6Capsicum. The method of claim 5 , wherein the genus plant is a ghost chili plant.7. The method of any one of to claim 5 , wherein the CS comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 1.8. The method of claim 7 , wherein the CS comprises the amino ...

Provision of malonyl-coa in coryneform bacteria and method for producing polyphenoles and polyketides with coryneform bacteria

A coryneform bacteria cell with an increased provision of Malonyl-CoA compared to its archetype, wherein the regulation and/or expression of one or more of genes fasB, gltA, accBC and accD1, and/or the functionality of the enzyme encoded by each gene is modified in a targeted manner. The cell may have one or more targeted modifications, including reduced or eliminated functionality of the fatty acid synthase FasB, mutation or partial or complete deletion of the fatty acid synthase encoding gene fasB, and/or reduced functionality of the promoter operatively linked to the citrate synthase gene gtIA, among other targeted modifications.

KETOREDUCTASE POLYPEPTIDES FOR THE REDUCTION OF ACETOPHENONES

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. 1Lactobacillus. A method for stereoselectively reducing 2 ,6-dichloro-3′-fluoroacetophenone substrate , optionally substituted at one or more positions selected from the group consisting of 3′ , 4′ , and 5′ , to the corresponding substituted (S)-1-phenethanol , which comprises contacting the substrate with an engineered ketoreductase polypeptide under reaction conditions suitable for stereoselectively reducing or converting the substrate to the corresponding substituted (S)-1-phenethanol product , wherein the engineered ketoreductase polypeptide is derived from a wild-type ketoreductase comprising an amino acid sequence at least 90% identical to the reference sequence of SEQ ID NO: 2 , 4 , or 98; and wherein said ketoreductase is capable of stereoselectively reducing 2 ,6-dichloro-3′-fluoroacetophenone to (S)-1-(2 ,6-dichloro-3-fluorophenyl) ethanol.2. The method of claim 1 , wherein the (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol is formed in greater than 90% stereomeric excess.3. The method of claim 1 , wherein the (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol is formed in greater than 99% stereomeric excess.4. The method of claim 1 , wherein at least 95% of the substrate is reduced to the product in less than 24 hours when the method is conducted with at least 200 g/L of substrate and with less than 2 g/L of the polypeptide.5. The method of claim 1 , wherein the method is carried out with whole cells that express the engineered ketoreductase polypeptide claim 1 , or an extract or lysate of such cells.6. The method of claim 1 , wherein the ...

Method of Preparing Piceatannol Using Bacterial Cytochrome P450 and Composition Therefor

Provided is a method of preparing piceatannol, and more particularly, to a method of preparing piceatannol from resveratrol using bacterial cytochrome P450 BM3 (CYP102A1) or mutants thereof, and a composition and a kit therefor. 1B. megaterium.. A method of preparing piceatannol , the method comprising reacting at least one enzyme selected from a group consisting of wild-type CYP102A1 and mutants of CYP102A1 with resveratrol wherein CYP102A1 is a cytochrome P450 from2. The method of claim 1 , further comprising adding an NADPH-generating system.3. The method of claim 2 , wherein the NADPH-generating system comprises glucose 6-phosphate claim 2 , NADP+ claim 2 , and yeast glucose 6-phosphate.4. The method of claim 1 , wherein the resveratrol is trans-resveratrol.5. The method of claim 1 , wherein the mutants of CYP102A1 are prepared by substituting one or more amino acids of SEQ ID NO:16 from the group consisting of:{'sup': 'th', 'substituting 47amino acid arginine (R) of wild-type CYP102A1 with one amino acid selected from a group consisting of alanine, valine, leucine, isoleucine, proline, methionine, phenylalanine, and tryptophan,'}{'sup': 'st', 'substituting 51amino acid tyrosine (Y) of wild-type CYP102A1 with one amino acid selected from a group consisting of phenylalanine, alanine, valine, leucine, isoleucine, proline, methionine, tryptophan,'}{'sup': 'th', 'substituting 64amino acid glutamic acid (E) of wild-type CYP102A1 with one amino acid selected from a group consisting of glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine,'}{'sup': 'th', 'substituting 74amino acid alanine (A) of wild-type CYP102A1 with one amino acid selected from a group consisting of glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine,'}{'sup': 'st', 'substituting 81amino acid phenylalanine (F) of wild-type CYP102A1 with one amino acid selected from a group consisting of alanine, valine, leucine, isoleucine, proline, methionine, and ...

CHIMERIC ENZYMES FOR CONVERSION OF LIGNIN-DERIVED CHEMICALS

Disclosed herein are enzymes useful for the dealkylation of aromatic substrates, including the conversion of guaiacol or guaethol to catechol. Methods of converting aromatic substrates found in lignin-based feedstocks such as pyrolysis oil into products such as catechol are also disclosed. Also presented herein are methods for rapidly evolving and optimizing genetic regions. 1. An isolated DNA molecule encoding a chimeric enzyme comprising a cytochrome P450 polypeptide and a catechol 1 ,2-dioxygenase polypeptide.2Amycolatopsis. The isolated DNA molecule of claim 1 , wherein the chimeric enzyme comprises at least 90% of the amino acids of the cytochrome P450 polypeptide GcoA from sp. ATCC 39116.3Acinetobacter baylyi. The isolated DNA molecule of claim 2 , wherein the chimeric enzyme further comprises at least 90% of the amino acids of the catechol 1 claim 2 ,2-dioxygenase CatA from ADP1.4. The isolated DNA molecule of claim 1 , wherein the chimeric enzyme has an amino acid sequence at least 90% identical to SEQ ID NO:2 or SEQ ID NO:10.5. The isolated DNA molecule of claim 1 , wherein the chimeric enzyme has the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:10.6. The isolated DNA molecule of claim 1 , further comprising an exogenous promoter operably linked to the DNA molecule.7. An expression vector comprising the DNA molecule of .8. A host cell that expresses a recombinant polypeptide encoded by the DNA molecule of .9PseudomonasAcinetobacter.. The host cell of claim 8 , wherein the cell is from a strain of or10P. putida.. The host cell of claim 9 , wherein the cell is11. An isolated chimeric enzyme polypeptide encoded by the DNA molecule of .12. A method for removing an alkyl group from an aromatic substrate claim 1 , comprising contacting a material containing the aromatic substrate with a chimeric enzyme comprising a cytochrome P450 polypeptide and a catechol 1 claim 1 ,2-dioxygenase polypeptide to generate a dealkylation product.13. The method of claim 12 , ...

Mutants of unspecific peroxygenase with high monooxygenase activity and uses thereof

The invention relates to an unspecific peroxygenase of the Agrocybe aegerita fungus, obtained by means of directed molecular evolution to facilitate the functional expression thereof in an active, soluble and stable form. The peroxygenase described in the invention shows a significant improvement in the functional expression thereof, improved monooxygenase activity and reduced peroxidase activity, in relation to the monooxygenase and peroxidase activities showed by the unspecific wild-type peroxygenase of A. aegerita . The peroxygenase of the invention is useful in chemical processes, including industrial transformations such as the selective oxyfunctionalisation of carbon-hydrogen bonds of various organic compounds.

A COMPOSITION OF PHOTOAUTOTROPHIC MICROORGANISMS AND CHEMOHETEROTROPHIC MICROORGANISMS IN A BIOFILM

A composition of microorganisms, comprising 1. A composition of microorganisms in a biofilm , comprising:photoautotrophic microorganisms which produce oxygen by photosynthetic water oxidation andchemoheterotrophic microorganisms which respire oxygen,wherein the photoautotrophic microorganisms and the chemoheterotrophic microorganisms are comprised in the biofilm, the biofilm further comprising components which were secreted by the photoautotrophic microorganisms and/or the chemoheterotrophic microorganisms.2. The composition of claim 1 , wherein the photoautotrophic microorganisms and/or the chemoheterotrophic microorganisms are capable of catalyzing the conversion of a substrate into a product.3. The composition of claim 2 , wherein in case of the chemoheterotrophic microorganisms the substrate is a substrate which is not naturally metabolized by the chemoheterotrophic microorganisms claim 2 , wherein the ability of converting the substrate was introduced into the chemoheterotrophic microorganisms by genetic modification.4. The composition of claim 2 , wherein in case of the photoautotrophic microorganisms the substrate is a substrate which is not naturally metabolized by the photoautotrophic microorganisms claim 2 , wherein the ability of converting the substrate was introduced into the photoautotrophic microorganisms by genetic modification.5. The composition of claim 1 , wherein the photoautotrophic microorganisms are algae and/or cyanobacterium.6Synechocystis.. The composition of claim 1 , wherein the photoautotrophic microorganisms are from the genus7. The composition of claim 1 , wherein the chemoheterotrophic microorganisms are bacteria.8Pseudomonas.. The composition of claim 1 , wherein the chemoheterotrophic microorganisms are from the genus9. The composition of claim 1 , wherein the biofilm is adhered to a surface of a carrier.10. The composition of claim 1 , wherein the carrier is a flat carrier claim 1 , a tube or a capillary.11. The composition of ...

FUNCTIONAL EXPRESSION OF MONOOXYGENASES AND METHODS OF USE

Methods and compositions for the oxidation of short alkanes by engineered microorganisms expressing recombinant enzymes is described, along with methods of use. 1. A synthetic polynucleotide for a soluble diiron monooxygenase enzyme which can be expressed in a microorganism of interest , comprising at least one monooxygenase coding region encoding a diiron monooxygenase enzyme , the at least one monooxygenase coding region linked to at least one promoter which will function in the microorganism of interest.2. The synthetic polynucleotide of claim 1 , further comprising at least one protein folding chaperone coding region encoding at least one protein folding chaperone claim 1 , the at least one protein chaperone coding region linked to at least one promoter which will function in the microorganism of interest.3. The synthetic polynucleotide of claim 1 , further comprising at least one mutation claim 1 , wherein the at least one mutation increases specificity for a monooxygenase substrate and/or increases production of a chemical.4. The synthetic polynucleotide of claim 2 , further comprising at least one mutation claim 2 , wherein the at least one mutation increases specificity for a monooxygenase substrate and/or increases production of a chemical.5. The synthetic polynucleotide of or claim 2 , wherein the soluble diiron monooxygenase enzyme is at least 60% identical to SEQ ID NO: 7 or SEQ ID NO: 9 or SEQ ID NO: 11 or SEQ ID NO: 13 or SEQ ID NO: 58 or SEQ ID NO: 60 or SEQ ID NO: 87 or SEQ ID NO: 89 or SEQ ID NO: 91 or SEQ ID NO: 93 or SEQ ID NO: 95 or SEQ ID NO: 97 or SEQ ID NO: 99 or SEQ ID NO: 101 or SEQ ID NO: 103 or SEQ ID NO: 105 or SEQ ID NO: 107 or SEQ ID NO: 109 or SEQ ID NO: 111 or SEQ ID NO: 113 or SEQ ID NO: 115 or SEQ ID NO: 117 or SEQ ID NO: 143 or SEQ ID NO: 145 or SEQ ID NO: 147 or SEQ ID NO: 149 or SEQ ID NO: 151 or SEQ ID NO: 153.6. The synthetic polynucleotide of or claim 2 , wherein the at least one mutation is a E to N at position 240 in SEQ ID ...

Compositions Having Dicamba Decarboxylase Activity and Methods of Use

Compositions and methods comprising polynucleotides and polypeptides having dicamba decarboxylase activity are provided. Further provided are nucleic acid constructs, host cells, plants, plant cells, explants, seeds and grain having the dicamba decarboxylase sequences. Various methods of employing the dicamba decarboxylase sequences are provided. Such methods include, for example, methods for decarboxylating an auxin-analog, method for producing an auxin-analog tolerant plant, plant cell, explant or seed and methods of controlling weeds in a field containing a crop employing the plants and/or seeds disclosed herein. Methods are also provided to identify additional dicamba decarboxylase variants. 1. A recombinant polypeptide having dicamba decarboxylase activity comprising:(a) an amino acid sequence having a similarity score of at least 548 for any one of SEQ ID NO: 51, 89, 79, 81, 95, or 100, wherein the similarity score is generated using the BLAST alignment program, with the BLOSUM62 substitution matrix, a gap existence penalty of 11, and a gap extension penalty of 1;(b) an amino acid sequence having at least 60%, 70%, 75%, 80% 90%, 95% or 100% sequence identity to any one of SEQ ID NOS: 1, 2, 4, 5, 16, 19, 21, 22, 26, 28, 30, 21, 32, 33, 34, 35, 36, 41, 43, 44, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 79, 81, 87, 88, 89, 91, 108, 109, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, or 129; or,(c) an amino acid sequence having at least 60%, 70%, 75%, 80% 90%, or 95% sequence identity to SEQ ID NO: 1, 2, 4, 5, 16, 19, 21, 22, 26, 28, 30, 21, 32, 33, 34, 35, 36, 41, 43, 44, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 79, 81, 87, 88, 89, 91, 108, 109, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, or 129;wherein the recombinant polypeptide comprises an active site having a catalytic residue geometry as set forth in Table 3 or having a substantially similar ...

HYDROXYSTILBENE PRODUCTION METHOD

The present invention is directed to provide novel methods of producing hydroxystilbene. According to the present invention, as a method of producing a compound having the general formula (II), 3. The method according to claim 1 , wherein Ris —H.4. The method according to claim 3 , wherein the compound having the general formula (II) is piceatannol.5. The method according to claim 1 , wherein the HPAH is a recombinant protein or a natural protein.6. The method according to claim 1 , wherein at least one selected from the group consisting of a purified HPAH recombinant protein claim 1 , a purified HPAH natural protein claim 1 , an extract from a bacterium expressing an endogenous HPAH claim 1 , an extract from a microorganism expressing an exogenous HPAH claim 1 , a bacterium expressing an endogenous HPAH claim 1 , and a microorganism expressing an exogenous HPAH is administered to the compound that is a substrate.78.-. (canceled)9. The method according to claim 6 , wherein the bacterium expressing an endogenous HPAH or the microorganism expressing an exogenous HPAH is administered to the compound in the presence of a surfactant.1013.-. (canceled)14. The method according to claim 2 , wherein Ris —H.15. The method according to claim 14 , wherein the compound having the general formula (II) is piceatannol.16. The method according to claim 14 , wherein the HPAH is a recombinant protein or a natural protein.17. The method according to claim 14 , wherein at least one selected from the group consisting of a purified HPAH recombinant protein claim 14 , a purified HPAH natural protein claim 14 , an extract from a bacterium expressing an endogenous HPAH claim 14 , an extract from a microorganism expressing an exogenous HPAH claim 14 , a bacterium expressing an endogenous HPAH and a microorganism expressing an exogenous HPAH is administered to the compound that is a substrate.18. The method according to claim 17 , wherein the bacterium expressing an endogenous HPAH or the ...

STILBENOID PRENYLTRANSFERASES FROM PLANTS

The process and system led to the identification of prenyltransferase genes from elicitor-treated peanut hairy roots. One of the prenyltransferases, AhR4DT-1 catalyzes a key reaction involved in the biosynthesis of prenylated stilbenoids, in which resveratrol is prenylated at its C-4 position to form arachidin-2, while another, AhR3′DT-1, was able to add the prenyl group to C-3′ of resveratrol. Each of these prenyltransferases has a high specificity for stilbenoid substrates, and their subcellular location in the plastid was confirmed by fluorescence microscopy. Structure analysis of the prenylated stilbenoids suggest that these two prenyltransferase activities represent the first committed steps in the biosynthesis of a large number of prenylated stilbenoids and their derivatives in peanut. 1. An isolated or purified nucleic acid molecule comprising:a gene sequence that encodes a polypeptide having stilbenoid prenyltransferase activity.2. The molecule of wherein the gene sequence is AhR4DT-1.3. The molecule of wherein the gene sequence is AhR3′DT-1.4. The molecule of wherein the gene sequence is AhR3′DT-2.5. The molecule of wherein the gene sequence is AhR3′DT-3.6. The molecule of wherein the gene sequence is AhR3′DT-4.7. A method of producing a prenylated stilbenoid in an organism claim 1 , cell or tissue claim 1 , the method comprising:expressing a gene sequence that encodes a polypeptide having stilbenoid prenyltransferase activity.8. The method of wherein the gene sequence that encodes a polypeptide having stilbenoid prenyltransferase activity is over-expressed.9. The method of wherein the gene sequence is AhR4DT-1.10. The method of wherein the gene sequence is AhR3′DT-1.11. The method of wherein the gene sequence is AhR3′DT-2.12. The method of wherein the gene sequence is AhR3′DT-3.13. The method of wherein the gene sequence is AhR3′DT-4.14. The method of wherein the organism is peanut.15. The method of wherein the organism is tobacco.16. The method of wherein ...

PROCESS AND COMPOSITION

The present invention relates to a process for recovering and purifying resveratrol produced by microbial fermentation. More particularly, the present invention relates to a process for recovering and purifying resveratrol produced by yeast fermentation. 1. A process for recovering and purifying resveratrol from a microbial fermentation broth said process comprising:(a) increasing the pH of the fermentation broth to about 10 to about 12;(b) separating and removing the host microbes such that a substantially microbe free liquid remains;(c) decreasing the pH of the substantially microbe free liquid such that crude resveratrol is precipitated;(d) separating the precipitated crude resveratrol;(e) dissolving the crude resveratrol recovered in step (c) in a purification solvent to form a crude resveratrol containing solution;(f) contacting the solution produced in step (e) with one or more absorbents; and(g) crystallizing the purified resveratrol from the purification solvent, said purified resveratrol having a lower concentration of impurities including bisnoryangonin than the crystals separated in step (c).2. A process according to where the one or more absorbents are selected from carbon claim 1 , optionally activated carbon claim 1 , ion exchange resins and/or AlO.3. A process according to where step (f) comprises the steps of:{'sub': 2', '3, 'i) contacting the solution produced in step (e) with one or more absorbents selected from ion exchange resins and/or AlO; and'}ii) contacting the solution produced in step i) with carbon, optionally activated carbon.4. A process according to any one of to where the pH to which the fermentation broth is adjusted in step (a) is about 11.5. A process according to any one of to wherein the host microbes are removed by ultrafiltration claim 1 , pressure filtration claim 1 , vacuum drum filtration claim 1 , and cross-flow filtration.6. A process according to wherein the host microbes are removed by ultrafiltration using an ultrafilter ...

ENZYMES AND METHODS FOR PROCESSING LIGNIN AND OTHER AROMATIC COMPOUNDS

Enzymes for depolymerizing lignin. The enzymes include dehydrogenases, β-etherases, and glutathione lyases. The dehydrogenases can comprise one or more or LigD, LigO, LigN, and LigL. The β-etherases can comprise one or more of LigE, LigF, LigP, and BaeA. The glutathione lyases can comprise any one or more of LigG and a number of non-stereospecific, optionally recombinant glutathione lyases derived from Sphingobium sp. SYK-6, , and other microorganisms. The enzymes can be combined in compositions and/or used in methods of processing lignin or other aromatic compounds in vitro. 1. A method of processing lignin , comprising contacting lignin comprising β-O-4 ether linkages in vitro with:a dehydrogenase comprising at least one of LigD, LigO, LigN, and LigL;a β-etherase comprising at least one of LigE, LigF, LigP, and an enzyme comprising a first polypeptide having an amino acid sequence of SEQ ID NO:40 or an amino acid sequence at least about 95% identical thereto and a second polypeptide having an amino acid sequence of SEQ ID NO:42 or an amino acid sequence at least about 95% identical thereto; and [{'sub': 'Nu', 'SEQ ID NO:18 (NaGST);'}, {'sub': 'Nu', 'residues 21-313 of SEQ ID NO:20 (recombinant NaGST);'}, {'sub': 'Nu', 'SEQ ID NO:22 (SYK6GST);'}, {'sub': 'Nu', 'residues 21-324 of SEQ ID NO:24 (recombinant SYK6GST);'}, 'SEQ ID NO:26 (ecYghU);', 'residues 21-313 of SEQ ID NO:28 (recombinant ecYghU);', 'SEQ ID NO:30 (ecYfcG);', 'SEQ ID NO:32 (ssYghU);', 'SEQ ID NO:34 (GST3); and', 'SEQ ID NO:36 (PcUre2pB1)., 'a glutathione lyase comprising any one or more of LigG and a non-stereospecific glutathione lyase comprising an amino acid sequence at least about 80% identical to any of2. The method of claim 1 , wherein the glutathione lyase comprises the non-stereospecific glutathione lyase.3. The method of claim 1 , wherein the glutathione lyase comprises the non-stereospecific glutathione lyase and the non-stereospecific glutathione lyase comprises an amino acid sequence at ...

IMMOBILIZED KETOREDUCTASES AND PROCESS FOR MAKING AND USING IMMOBILIZED KETOREDUCTASE

The invention is directed to immobilized ketoreductases and methods of making and using them. Enzymes are protein molecules which serve to accelerate the chemical reactions of living cells (often by several orders of magnitude). Without enzymes, most biochemical reactions would be too slow to even carry out life processes. Enzymes display great specificity and are not permanently modified by their participation in reactions. Since they are not changed during the reactions, enzymes can be cost effectively used as catalysts for a desired chemical transformation. 1. An immobilized ketoreductase comprising:a recombinant ketoreductase; anda resin, wherein the recombinant ketoreductase is attached to the resin and wherein the immobilized ketoreductase is stable in a solvent system that comprises at least 90% of organic solvents.2. An immobilized ketoreductase of wherein the immobilized ketoreductase is stable in a solvent system that comprises at least 95% of organic solvents.3. An immobilized ketoreductase of wherein the recombinant ketoreductase is attached to a resin by adsorption.4. An immobilized ketoreductase of wherein the recombinant ketoreductase is attached to a resin by covalent bonds.5. An immobilized ketoreductase of wherein the recombinant ketoreductase is attached to a resin by ionic bonds.6. An immobilized ketoreductase of wherein the resin comprises polymethacrylate with epoxide functional groups claim 1 , polymethacrylate with amino epoxide functional groups claim 1 , styrene/DVB copolymer or polymethacrylate with octadecyl functional groups claim 1 , silica claim 1 , carboxylic group or quaternary ammonia.7. An immobilized ketoreductase of wherein the resin comprises styrene/DVB copolymer claim 3 , silica or polymethacrylate with octadecyl functional groups.8. An immobilized ketoreductase of wherein the resin is selected from the group consisting of: LEWATIT VPOC 1600 claim 3 , SEPABEAD EXE120 claim 3 , DIAION HP2MG claim 3 , IMMOBEAD-EC1 claim 3 , ...

CHEMICAL ENGINEERING PROCESSES AND APPARATUS FOR THE SYNTHESIS OF COMPOUNDS

The present invention provides methods for producing cannabinoids and cannabinoid analogs as well as a system for producing these compounds. The inventive method is directed to contacting a compound according to Formula I or Formula II with a cannabinoid synthase. 1. A system for producing cannabinoid products , comprising:a fermentor holding a cell culture medium comprising genetically engineered yeast or bacterial cells that are (a) producing a cannabinoid synthase selected from the group consisting of tetrahydrocannabinolic acid (THCA) synthase and cannabidiolic acid (CBDA) synthase and (b) secreting the cannabinoid synthase into the culture medium; anda bioreactor containing a reaction mixture comprising the cannabinoid synthase and 3-(3,7-dimethylocta-2,6-diene-1-yl)-2,4-dihydroxy-6-propyl-1-benzoic acid (CBGVA) and configured to form one or more cannabinoid products.2. The system of claim 1 , further comprising a control mechanism configured to control one or more properties of the reaction mixture so as to modify the amount of the one or more cannabinoid products claim 1 , wherein the one or more properties of the reaction mixture are selected from the group consisting of ionic strength claim 1 , pH claim 1 , temperature claim 1 , and pressure.3. The system of claim 2 , wherein the property of the reaction mixture is pH.4. The system of claim 3 , wherein the pH is from 4 to less than 8.5. The system of claim 4 , wherein the pH is between 4.0 and 6.0.6. The system of claim 2 , wherein the control mechanism comprises a processing circuit and a display to control the one or more properties of the reaction mixture.7. The system of claim 1 , further comprising a control mechanism configured to regulate a property of the culture medium containing the cannabinoid synthase claim 1 , wherein the property of the culture medium containing the cannabinoid synthase is one or more of oxygen level of the culture medium claim 1 , rate of agitation of the culture medium claim ...

Host Cells and Methods for Oxidizing Aromatic Amino Acids

The present invention provides for a method of producing an oxidation product of an aromatic amino acid in a genetically modified host cell. The method comprises culturing the genetically modified host cell under a suitable condition such that the culturing results in the genetically modified host cell producing oxidation product of an aromatic amino acid. The host cell comprises an enzyme capable of catalyzing the oxidation of aromatic amino acid. In some embodiments, the host cell is capable of biosynthesizing BH4 or MH4 from GTP. 1. A method of producing one or more oxidation products of an aromatic amino acid in a genetically modified host cell comprising: (a) culturing the genetically modified host cell under a suitable condition such that the culturing results in the genetically modified host cell producing one or more oxidation products of an aromatic amino acid.2. The method of claim 1 , wherein the host cell comprises an enzyme capable of catalyzing the oxidation of aromatic amino acid.3. The method of claim 1 , wherein the aromatic amino acid is tyrosine or tryptophan.4. The method of claim 3 , wherein the aromatic amino acid is tyrosine and the one or more oxidation products are L-DOPA claim 3 , dopamine claim 3 , 3 claim 3 ,4-dihydroxyphenylacetaldehyde claim 3 , 3 claim 3 ,4-dihydroxypehylethanol (hydroxytyrosol) claim 3 , reticuline claim 3 , thebaine claim 3 , and/or morphine.5. The method of claim 4 , wherein the host cell is capable of biosynthesizing BH4 or MH4 from GTP.6. The method of claim 3 , wherein the aromatic amino acid is tryptophan and one or more oxidation products are 5-hydroxytryptophan claim 3 , serotonin claim 3 , and/or melatonin.7. The method of claim 6 , wherein the host cell is capable of biosynthesizing BH4 or MH4 from GTP.8. A genetically modified host cell capable of producing one or more oxidation products of an aromatic amino acid.9. The host cell of claim 8 , wherein the aromatic amino acid is tyrosine.10. The host cell of ...

METHOD FOR PREPARING p-VINYL PHENOLS

A biocatalytic method is provided for preparing p-vinyl phenols by a three-step, one-pot reaction according to the following reaction scheme: 110.-. (canceled)12. The method according to claim 11 , wherein the reaction is performed at a pH of about 8 to 9.13. The method according to claim 12 , wherein the reaction is performed at a pH of about 8.14. The method according to claim 11 , wherein in step (b) the catalytic action uses a catalyst comprising a tyrosine ammonia-lyase (TAL).15. The method according to claim 14 , wherein the tyrosine ammonia-lyase (TAL) is used in a form of whole cells containing recombinant enzyme.16. The method according to claim 11 , wherein in step (c) the catalytic action uses a catalyst comprising a ferulic acid decarboxylase (FAD).17. The method according to claim 16 , wherein the ferulic acid decarboxylase (FAD) is used in a form of whole cells containing recombinant enzyme.18. The method according to claim 11 , wherein the one-pot reaction is performed in an aqueous buffer system in a presence of a water-immiscible co-solvent.19. The method according to claim 18 , wherein the co-solvent comprises diethyl ether.20. The method according to claim 19 , wherein the diethyl ether is present in an amount of 5% (v/v) based on the aqueous buffer system.21. The method according to claim 11 , wherein the phenol (1) is substituted in at least one of ortho- and meta-positions with at least one substituent R selected from halogens claim 11 , Calkyl claim 11 , and Calkoxy. The invention relates to a biocatalytic method for preparing p-vinyl phenols.Vinyl phenol derivatives serve as useful components in polymer chemistry and may be used, for example, for forming dielectric layers in the production of chemical and biological sensors. Halogenated derivatives are used, for example, for producing flame retardants as well as chalcones, a well-known class of organic compounds having a wide range of biological activities.A selective para-vinylation of non- ...

METHOD FOR THE ENZYMATIC CONVERSION OF A PHENOL SUBSTRATE INTO A CORRESPONDING CATECHOL PRODUCT

A method for the enzymatic conversion of a phenol substrate into a corresponding catechol product comprises the step of incubating the phenol substrate with a tyrosinase enzyme, or a functional derivative thereof, in a reaction mixture, for a period of time sufficient to allow the enzyme convert at least some of the phenol substrate into the catechol product. 1Ralstonia solanacearum. A method for the enzymatic conversion of a halophenol into a halocatechol , the method comprising the steps of incubating the halophenol substrate with a tyrosinase enzyme , or a functional derivative thereof , in a reaction mixture , in which the reaction mixture comprises ascorbic acid or its sodium salt , for a period of time sufficient to allow the enzyme convert at least some of the halophenol substrate into the halocatechol product.2. A method according to in which the period of time is sufficient to allow the enzyme to convert at least 90% of the halophenol into a halocatechol.3. A method according to in which the reaction mixture comprises at least 10 mM of the halophenol substrate.4. A method according to in which the reaction mixture comprises at least 20 mM ascorbic acid.5. A method according to claim 1 , in which the halophenol is 3-halophenol or 4-halophenol claim 1 , and the product is a 3-halocatechol or 4-halocatechol claim 1 , respectively.6Ralstonia solanacearumRalstonia solanacearum. A method according to in which the functional derivative of the tyrosinase enzyme is an engineered variant having 1 to 5 amino acid alterations compared with the tyrosinase enzyme claim 1 , wherein the or each alternation is selected from insertion claim 1 , additional claim 1 , deletion and substitution of an amino acid.7Ralstonia solanacearum. A method according to in which the functional derivative of the tyrosinase enzyme is selected from the group consisting of Y119F claim 1 , V153A claim 1 , D317Y and L330V (RV145); T1831 claim 1 , F185Y claim 1 , N322S claim 1 , and T359M (RVC10); ...

Mutant enzymes

This invention relates to mutant enzymes with enhanced properties and processes for oxidation of organic compound substrates using such enzymes.

Isolated Codon Sequence

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 1. A method for producing enzymes and products , wherein the method comprises:inserting a gene into an expression system and thereby yielding a modified host;optimizing the modified host and thereby yielding the enzymes and the products;purifying the enzymes and products; andwherein the gene contains a codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleic acid sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34; or an codon-optimized nucleic acid sequence of nucleic acids 5052 to 6671 of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acids 5052 to 6671 of SEQ ID NO: 34.2Cannabis. The method of claim 1 , wherein the products are found in a plant.3. The method of claim 1 , wherein the products comprise at least one of: cannabinoids claim 1 , terpenoids claim 1 , stilbenoids claim 1 , flavonoids claim 1 , phenolic amides claim 1 ...

Isolated Codon Sequence

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 1. A host organism , wherein the host organism comprises:the host organism that is in a modified form in comparison to a natural state of the host organism via gene insertion, and thereby contains an increased yield of product in comparison to a yield of product in the natural state of the host organism;a plurality of pathways in the host organism, wherein a first pathway of the plurality of pathways generates the product;a plurality of substrates for deriving the product; anda codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleic acid sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34;2. The host organism of claim 2 , wherein the host organism is a bacteria claim 2 , plant claim 2 , alga claim 2 , or fungus.3S. cerevisiae, E. coli, P. pastorisis, N. crassa, S. pombe, R. palmatum, C. Tonga, O. sativa, A. arborescens, A. terrus, C. sativa, S. griseus, S. erythera, S. coelicolor, S. toxytruini, S. cellulosum, P. ...

SOLID FORMS OF MENAQUINOLS

Disclosed are solid forms of menaquinol and processes for obtaining them by chemical or enzymatic reduction of menaquinone. Said solid forms possess high stability to oxidation which allows effective use of menaquinol in the most common formulations wherein vitamin K2 is used. 2. The process of wherein step a) is performed at a temperature between 2° C. and 75° C.3. The process of wherein the solution of sodium dithionite has a pH ranging between 3 and 8.4. The process of claim 1 , further comprising washing of the crystalline form obtained in step c) with a solution of ascorbic acid followed by drying of the crystals at a temperature not exceeding 40° C.5. The process of wherein claim 1 , in the compound of formula (II) claim 1 , n is 6.6. The process of wherein claim 1 , in the compound of formula (II) claim 1 , n is 3.7. A crystalline form of menaquinol 7 having a DSC profile as shown in claim 1 , showing an endothermic peak at a temperature of 75° C. or higher claim 1 , and an FT-IR spectrum as reported in claim 1 , having a broad peak at 3340-3350 cm claim 1 , three characteristic peaks at 2964 claim 1 , 2916 claim 1 , and 2851 cm claim 1 , and sharp peaks at 1599 claim 1 , 1504 claim 1 , 1326 claim 1 , 1182 claim 1 , 1149 claim 1 , 1096 claim 1 , 1046 claim 1 , 980 claim 1 , 950 claim 1 , and 752 cm.8. A crystalline form of menaquinol 7 having a DSC profile as shown in claim 1 , showing an endothermic peak at a temperature of 78.4° C. or higher and a second endothermic event in the range of 39-45° C.10BacillusBacillus subtilis, Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus megaterium, Bacillus pumilusBacillus licheniformis.. The process of wherein the reducing step is performed using a micro-organism of the genus claim 9 , selected from the group consisting of claim 9 , and11PseudomonasEscherichiaEnterobacter.. The process of wherein the reducing step is performed using a micro-organism selected from the group consisting of a micro-organism ...

BIOSYNTHESIS OF CANNABINOIDS AND CANNABINOID PRECURSORS

Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro. 171-. (canceled)72. A method for producing a cannabinoid or cannabinoid precursor comprising culturing a host cell that comprises a heterologous polynucleotide encoding a polyketide synthase (PKS) , wherein the PKS comprises a sequence that is at least 95% identical to SEQ ID NO: 7.73. The method of claim 72 , wherein relative to the sequence of SEQ ID NO: 7 claim 72 , the PKS comprises an amino acid substitution at a residue corresponding to position 28 claim 72 , 34 claim 72 , 50 claim 72 , 70 claim 72 , 71 claim 72 , 76 claim 72 , 88 claim 72 , 100 claim 72 , 151 claim 72 , 203 claim 72 , 219 claim 72 , 285 claim 72 , 359 claim 72 , and/or 385 in SEQ ID NO: 7.74. The method of claim 72 , wherein the PKS comprises:a) the amino acid P at a residue corresponding to position 28 in SEQ ID NO: 7;b) the amino acid Q at a residue corresponding to position 34 in SEQ ID NO: 7;c) the amino acid N at a residue corresponding to position 50 in SEQ ID NO: 7;d) the amino acid M at a residue corresponding to position 70 in SEQ ID NO: 7;e) the amino acid Y at a residue corresponding to position 71 in SEQ ID NO: 7;f) the amino acid I at a residue corresponding to position 76 in SEQ ID NO: 7;g) the amino acid A at a residue corresponding to position 88 in SEQ ID NO: 7;h) the amino acid P or T at a residue corresponding to position 100 in SEQ ID NO: 7;i) the amino acid P at a residue corresponding to position 151 in SEQ ID NO: 7;j) the amino acid K at a residue corresponding to position 203 in SEQ ID NO: 7;k) the amino acid C at a residue corresponding to position 219 in SEQ ID NO: 7;l) the amino acid A at a residue corresponding to position 285 in SEQ ID NO: 7;m) the amino acid M at a residue corresponding to position 359 in SEQ ID NO: 7; and/orn) the amino acid M at a residue corresponding to position 385 in SEQ ID NO: 7.75. The method of claim 72 , wherein ...

METHOD TO INCREASE THE YIELD OF PRODUCTS IN PLANT MATERIAL

A method to increase the production of products of interest in plant material including plant cultures, such as, for example, cell suspension cultures, root cultures, and hairy root cultures is provided. In one embodiment, the method is to contacting the plant material with a precursor or xenobiotic when producing a product of interest from a plant. In another embodiment the plant material is also contacted with a trapping agent. The process may also provide for contacting an elicitor of the product of interest with the plant material. An embodiment provides for contacting an elicitor, precursor and trapping agent with the plant material. The ability to produce novel compounds such as glucosides and glucuronides is provided. 1. A method of increasing the production of a product of interest from plant material , the method comprising identifying a product of interest , providing plant material capable of producing said product of interest , contacting said plant material with a precursor of said product of interest and producing an increased amount of said product of interest compared to said method in which said precursor is not contacted with said plant material.2. The method of claim 1 , further comprising contacting said plant material with a trapping agent.3. The method of claim 1 , further comprising contacting said plant material with an elicitor.4. The method of claim 1 , further comprising contacting said plant material with a trapping agent and an elicitor.5. The method of claim 1 , wherein said product of interest is selected from a phenolic claim 1 , alkaloid or terpenoid compound.6. The method of claim 1 , wherein said product of interest is selected from a stilbenoid claim 1 , flavonoid or alkaloid.7. The method of claim 1 , wherein said precursor comprises chyrsin or naringenin or squalene.8. The method of claim 1 , wherein said product of interest is a stilbenoid claim 1 , said precursor comprises piceatannol and further comprising contacting said ...

Optimized microbial cells for production of melatonin and other compounds

Described herein are recombinant microbial host cells comprising biosynthetic pathways and their use in producing oxidation products and downstream products, e.g., melatonin and related compounds, as well as enzyme variants, nucleic acids, vectors and methods useful for preparing and using such cells. In specific aspects, the present invention relates to monooxygenases, e.g., amino acid hydroxylases, with a modified cofactor-dependency, and to enzyme variants and microbial cells providing for an improved supply of cofactors. 2. The variant of claim 1 , wherein(i) the mutation in the residue corresponding to residue E147 is an amino acid substitution selected from 147K, 147R and 147H;(ii) the mutation in the residue corresponding to residue N242 is the amino acid substitution 242I; and(iii) the mutation in the residue corresponding to P244 is selected from 244C, 244D, 244L and 244Q.3. The variant of claim 2 , wherein the mutation in the residue corresponding to E147 is E147K claim 2 , the mutation in the residue corresponding to N242 is N242I claim 2 , and the mutation in the residue corresponding to P244 is P244C.4Homo sapiens. The variant of claim 3 , comprising a segment which has at least 95% sequence identity to the segment corresponding to residues E147 to T460 of TPH having the sequence of SEQ ID NO:3.5. The variant of claim 1 , which provides for a tryptophan hydroxylation activity which is at least 110% of that of the native or parent TPH.6. A nucleic acid sequence encoding the variant TPH of .7. A recombinant microbial cell comprising the nucleic acid sequence of .8. A TPH having the sequence of SEQ ID NO:13 except for E2K claim 6 , N97I and P99C mutations.9. A nucleic acid sequence encoding the TPH of .10. A recombinant microbial cell comprising the nucleic acid sequence of .11E. coli. The recombinant microbial cell of claim 10 , further comprising nucleic acid sequences encoding a pterin-4a-carbolamine dehydratase (PCD) and a GTP cyclohydrolase I (GCH1) ...

METHOD FOR MASS-PRODUCING VINIFERIN USING STEVIOSIDE FROM CELL CULTURE OF GRAPEVINE TISSUE

The present invention relates to a method for mass-production of viniferin using stevioside from cell culture of grape tree tissue. Viniferin is known to be effective for protection of liver, anticancer, antioxidant, and skin whitening, have an effect of inhibiting oxidation of low-density lipoprotein and high-density lipoprotein and inhibiting the proliferation and migration of vascular smooth muscle cells. Therefore, the present invention is very useful for the mass production of viniferin among the useful substances (stilbene compounds) from a callus derived from the anther tissue of the grape plant, which is very important for the related industries. 1. A method of mass production of viniferin derived from a grape tree , the method comprising the steps of:(a) transplanting a grape tree tissue fragment on a callus induction medium to induce a grape tree callus;(b) culturing the induced grape tree callus in a growth medium to grow; and(c) adding an activity-inducing agent and a solubilizing agent to the grown callus, followed by shaking culture.2. The method according to claim 1 , wherein the activity-inducing agent and the solubilizing agent in step (c) are methyl jasmonate and stevioside claim 1 , respectively.3. The method according to claim 2 , wherein the methyl jasmonate and stevio side are added at a concentration of 50 μM to 350 μM and 40 mM to 60 mM claim 2 , respectively.4. The method according to claim 1 , wherein the viniferin is delta (δ)-viniferin or epsilon (ε)-viniferin.5. A composition for mass production of viniferin from a grape tree callus claim 1 , the composition comprising methyl jasmonate (MeJA) and stevioside as active ingredients.6. The composition according to claim 5 , the composition comprising 50 μM to 350 μM methyl jasmonate and 40 mM to 60 mM stevioside.7. A method of mass production of resveratrol derived from a grape tree claim 5 , the method comprising the steps of:(a) transplanting a grape tree tissue fragment on a callus ...

Modified Plants for Producing Aroma/Fine/Specialty Chemicals

The present disclosure describes genetically modified plants that contain one or more exogenous genes associated with aroma/fine/specialty compound biosynthesis, which are capable of producing aroma/fine/specialty compounds. 182-. (canceled)83. A modified plant comprising one or more exogenous polynucleotides encoding one or more enzymes associated with an aroma compound biosynthesis , a fine compound biosynthesis , a specialty compound biosynthesis or a combination thereof.84. The modified plant of claim 83 , wherein the one or more enzymes is a synthase claim 83 , a reductase claim 83 , a transferase claim 83 , or a combination thereof.85. The modified plant of claim 84 , whereinthe synthase is a phenylacetaldehyde synthase, a propenylphenol synthase, an allylphenol synthase, a biologically active fragment or variant thereof or a combination thereof;the reductase is a phenylacetaldehyde reductase or a biologically active fragment or variant thereof; andthe transferase is an acyl transferase or a biologically active fragment or variant thereof86Larrea tridentateLarrea tridentata. The modified plant of claim 85 , wherein the propenyl phenol synthase is propenylphenol synthase or a biologically active fragment or variant thereof and the allylphenol synthase is allylphenol synthase or a biologically active fragment or variant thereof.87. The modified plant of claim 83 , wherein the aroma compounds include at least one or more metabolites associated with 2-phenyl ethanol synthesis.88. The modified plant of claim 83 , wherein the modified plant produces a 2-phenyl ethanol claim 83 , an allyl phenol claim 83 , a propenyl phenol or a combination thereof.89. The modified plant of claim 83 , wherein the modified plant is selected from the group consisting of a poplar claim 83 , a poplar hybrid claim 83 , an aspen claim 83 , and a red alder.90. The modified plant of claim 83 , wherein the modified plant produces aroma compounds that are sequestered in the modified plant as ...

Polypeptides Having Peroxygenase Activity

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 143-. (canceled)44. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted , linear or branched , alkane or saturated fatty acid having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3 , comprising contacting the alkane or saturated fatty acid with hydrogen peroxide and a polypeptide having peroxygenase activity and at least 95% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2 , wherein the alkane or saturated fatty acid is optionally substituted with one or two substituents selected from the group consisting of halogen , hydroxyl , carboxyl , amino , nitro , cyano , thiol , sulphonyl , formyl , acetyl , methoxy , ethoxy , phenyl , benzyl , xylyl , carbamoyl and sulfamoyl.45. The method of claim 44 , wherein an alkane is hydroxylated.46. The method of claim 45 , wherein the alkane is pentane claim 45 , hexane claim 45 , heptane claim 45 , octane claim 45 , nonane claim 45 , decane claim 45 , undecane claim 45 , dodecane claim 45 , tridecane claim 45 , tetradecane claim 45 , pentadecane or hexadecane claim 45 , or an isomer thereof.47. The method of claim 45 , wherein the alkane is unsubstituted.48. The method of claim 45 , wherein the alkane is linear.49. The method of claim 45 , wherein the alkane is converted to a diol by introduction of two hydroxy groups.50. The method of claim 44 , wherein a saturated fatty acid is hydroxylated.51. The method of claim 50 , wherein the saturated fatty acid is selected from the group consisting of butanoic acid claim 50 , pentanoic acid claim 50 , hexanoic acid claim 50 , heptanoic acid claim 50 , octanoic acid claim 50 , nonanoic acid ...

Polypeptides Having Peroxygenase Activity

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 143-. (canceled)44. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted , linear or branched , alkane or saturated fatty acid having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3 , comprising contacting the alkane or saturated fatty acid with hydrogen peroxide and a polypeptide having peroxygenase activity and at least 95% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2 , wherein the alkane or saturated fatty acid is optionally substituted with one or two substituents selected from the group consisting of halogen , hydroxyl , carboxyl , amino , nitro , cyano , thiol , sulphonyl , formyl , acetyl , methoxy , ethoxy , phenyl , benzyl , xylyl , carbamoyl and sulfamoyl.45. The method of claim 44 , wherein an alkane is hydroxylated.46. The method of claim 45 , wherein the alkane is pentane claim 45 , hexane claim 45 , heptane claim 45 , octane claim 45 , nonane claim 45 , decane claim 45 , undecane claim 45 , dodecane claim 45 , tridecane claim 45 , tetradecane claim 45 , pentadecane or hexadecane claim 45 , or an isomer thereof.47. The method of claim 45 , wherein the alkane is unsubstituted.48. The method of claim 45 , wherein the alkane is linear.49. The method of claim 45 , wherein the alkane is converted to a diol by introduction of two hydroxy groups.50. The method of claim 44 , wherein a saturated fatty acid is hydroxylated.51. The method of claim 50 , wherein the saturated fatty acid is selected from the group consisting of butanoic acid claim 50 , pentanoic acid claim 50 , hexanoic acid claim 50 , heptanoic acid claim 50 , octanoic acid claim 50 , nonanoic acid ...

PROCESS FOR FUNGAL MODIFICATION OF LIGNIN AND PREPARING WOOD ADHESIVES WITH THE MODIFIED LIGNIN AND WOOD COMPOSITES MADE FROM SUCH ADHESIVES

Disclosed herein are method to modify the lignin with particular fungal species, and procedure to synthesize phenolic adhesives with the modified lignin as raw materials, and the adhesives compositions and methods for making adhesive compositions, and methods for making lingo-cellulosic composites from renewable materials. Four fungi in examples are (Spreng.) Pat. (FTK 329A), (Bres.) Parmasto (FTK 332A), (Ellis & Everh.) Kotl. & Pouz. (FTK 76A) and (Pers.) Bondartsev (FTK 122B). Lignin used in examples are organosolv lignin, Kraft lignin, and ammonium lignosulfonate. The present invention includes methods to (1) modify of lignin with fungi; (2) in-situ polymerize modified lignin-phenol-formaldehyde to generate bio-modified lignin-phenol-formaldehyde adhesive in liquid form, and (3) manufacture composite panels with bio-modified lignin-phenol-formaldehyde resins. 1. A process for the preparation of polymer adhesives comprising the steps of:{'i': Lenzites elegans, Phanerochaete cremea, Pycnoporellus alboluteus', 'Meruliopsis taxicola, 'providing a fungal species selected from the group consisting of and and combinations thereof;'}providing a lignin;preparing each of the fungal species;preparing the lignin for fungal species modification of the lignin;mixing the prepared fungal species and the prepared lignin to produce a lignin/fungal species suspension;incubating the suspension;separating the suspension into a solid comprising the fungal species and a liquid comprising the modified lignin; andtreating the liquid to produce a reactive solid for the polymer adhesive.2. The process of claim 1 , further comprising:providing at least one phenolic compound, at least one formaldehyde compound, an alkali metal hydroxide and water;mixing the at least one phenolic compound, the at least one formaldehyde compound, the alkali metal hydroxide, the water and the reactive solid to produce a methylolation medium (at a pH of about 10 or less);maintaining the medium at a methylolation ...

RECOVERY, DECARBOXYLATION, AND PURIFICATION OF CANNABINOIDS FROM ENGINEERED CELL CULTURES

Methods of recovering cannabinoids from cell cultures include methods comprising steps of separating the cell culture at a temperature above the melting point of the cannabinoid to separate a light phase comprising liquid state cannabinoid from a heavy phase; and methods comprising treating the cell culture at a temperature below the melting point of the cannabinoid to separate a light phase from a heavy phase comprising solid state cannabinoid. Other methods include contacting the culture with a water-miscible solvent to form a water-miscible phase and an aqueous phase, separating the two phases and recovering the cannabinoid. Other methods include contacting the culture with a water-immiscible solvent to form a water-immiscible phase and an aqueous phase, separating the two phases, and recovering the cannabinoid. Other methods include washing the inner surface of a fermentation vessel with alkaline solution to recover cannabinoid attached to the vessel surface. Various methods make use of aqueous solvent systems comprising no organic solvent, aqueous solvent systems comprising added water-miscible organic solvent, and dual-phase aqueous/water-immiscible solvent systems. 133-. (canceled)34. A method of recovering a cannabinoid from a cell culture , the method comprising:a) providing a cell culture comprising cells engineered to produce a cannabinoid in a culture medium;b) treating the culture at a temperature below the melting point of the cannabinoid to generate a first pellet comprising cells, insoluble cellular material, and the cannabinoid, and a first supernatant comprising culture medium;c) removing the first supernatant from the first pellet;d) adding a water-immiscible organic solvent to the first pellet to generate a solvent-extracted pellet mixture;e) separating, the solvent-extracted pellet mixture to generate a heavy phase and a light phase, wherein the light phase comprises the water-immiscible organic solvent and the cannabinoid; andf) recovering the ...

PHLOROGLUCINOL-RESISTANT CELL, IN PARTICULAR YEAST

The present invention relates to a living cell, preferably a host cell, that is phloroglucinol resistant, said host cell being characterized in that it overexpresses a polypeptide selected from (i) transmembrane transporters of the ABC family, in particular transmembrane transporters of the PDR subfamily, (ii) transcription factors which control the expression of the transmembrane transporters of the PDR subfamily, and (iii) transmembrane transporters of the MFS family; preferably a polypeptide selected from SNQ2, STE6, PDR3, AQR1, DTR1, FLR1, QDR1, YHK8 and ATR1; and more preferably the SNQ2 transporter. The present invention also relates to a method for producing a phloroglucinol-resistant recombinant host cell. The present invention also relates to a method for producing phloroglucinol. 1. Living cell , preferably host cell , that is phloroglucinol resistant , characterized in that it withstands a phloroglucinol concentration , in its culture medium , of greater than or equal to 1 g·l.2. Living cell claim 1 , preferably host cell claim 1 , according to claim 1 , characterized in that it overexpresses at least one membrane transporter claim 1 , or at least one transcription factor which controls the expression of said membrane transporter.3. Living cell claim 1 , preferably host cell claim 1 , according to claim 1 , characterized in that said membrane transporter belongs to the ABC transporter family claim 1 , preferably to the PDR subfamily claim 1 , or to the MFS transporter family.4. Living cell claim 1 , preferably host cell claim 1 , according to claim 1 , characterized in that said transporter or said transcription factor is selected from SNQ2 claim 1 , STE6 claim 1 , PDR3 claim 1 , AQR1 claim 1 , DTR1 claim 1 , FLR1 claim 1 , QDR1 claim 1 , YHK8 and ATR1.5. Living cell claim 1 , preferably host cell claim 1 , according to claim 1 , characterized in that it withstands a phloroglucinol concentration claim 1 , in its culture medium claim 1 , of greater than or ...

METHOD FOR PRODUCING RETINOID FROM MICROORGANISM

The present invention relates to a method for producing retinoid from a microorganism, and more specifically, to a method for effectively obtaining retinoid, which lacks stability, from a microorganism by cultivating the microorganism capable of producing retinoid in a medium containing a lipophilic substance, and separating retinoid from the lipophilic substance. 1. A method for production of retinoid from a microorganism , comprising:culturing a microorganism having retinoid producing efficacy in a medium containing a lipophilic substance; andisolating the retinoid from the lipophilic substance.2. The method according to claim 1 , wherein the microorganism is bacteria claim 1 , fungi claim 1 , isolated animal cell or a combination thereof.3Escherichiabacilluscorynebacteriumkluyveromyces. The method according to claim 1 , wherein the microorganism is the genus claim 1 , the genus claim 1 , the genus claim 1 , yeast claim 1 , or a combination thereof.5. The method according to claim 1 , wherein the lipophilic substance is octane claim 1 , decane claim 1 , dodecane claim 1 , tetradecane claim 1 , phytosqualane claim 1 , mineral oil claim 1 , isopropyl myristate claim 1 , cetyl ethylhexanoate claim 1 , dioctanoyl decanoyl glycerol claim 1 , squalane claim 1 , or a combination thereof.6. The method according to claim 1 , wherein a ratio by volume of the medium to the lipophilic substance ranges from 1:0.2 to 3.0.7. The method according to claim 1 , wherein the culturing is performed while agitating.8. The method according to claim 1 , wherein the medium further includes glycerol.9. The method according to claim 1 , wherein the medium further includes glucose.10. The method according to claim 1 , wherein the isolating includes removing cells from a culture solution and then isolating the retinoid from dodecane.11. The method according to claim 1 , wherein the retinoid is at least one selected from a group consisting of retinal claim 1 , retinol claim 1 , retinyl ester ...

Production Of Enantiopure alpha-Hydroxy Carboxylic Acids From Alkenes By Cascade Biocatalysis

The invention provides compositions comprising an alkene epoxidase and a selective epoxide hydrolase, such as a recombinant microorganism comprising a first heterologous nucleic acid encoding an alkene epoxidase and a second heterologous nucleic acid encoding a selective epoxide hydrolase. Exemplary alkene epoxidases include StyAB, while exemplary selective epoxide hydrolases include epoxide hydrolases from Sphingomonas, Solanum tuberosum , or Aspergillus . The invention also provides non-toxic methods of making enantiomerically pure vicinal diols or enantiomerically pure alpha-hydroxy carboxylic acids using these compositions and microorganisms.

In vitro methods for processing lignin and other aromatic compounds

Enzymes for depolymerizing lignin. The enzymes include dehydrogenases, β-etherases, and glutathione lyases. The dehydrogenases can comprise one or more or LigD, LigO, LigN, and LigL. The β-etherases can comprise one or more of LigE, LigF, LigP, and BaeA. The glutathione lyases can comprise any one or more of LigG and a number of non-stereospecific, optionally recombinant glutathione lyases derived from Sphingobium sp. SYK-6, Novosphingobium aromaticivorans, Escherichia coli, Streptococcus sanguinis, Phanerochaete chrysosporium, and other microorganisms. The enzymes can be combined in compositions and/or used in methods of processing lignin or other aromatic compounds in vitro.

ENZYMES AND METHODS FOR DEALKYLATION OF SUBSTRATES

Disclosed herein are enzymes and organisms useful for the dealkylation of products derived from lignin depolymerization, including the conversion of guaiacol or guaethol to catechol or the conversion of anisole to phenol. Methods of converting guaiacol or guaethol to catechol or anisole to phenol using enzymes or organisms expressing the same are also disclosed. 1. A method for removing an alkyl group from an aromatic substrate , comprising contacting a material containing the aromatic substrate with a cytochrome P450 polypeptide and a reductase polypeptide to generate a dealkylation product.2. The method of claim 1 , wherein the contacting step comprises culturing a microorganism with the material containing the aromatic substrate claim 1 , wherein the microorganism expresses an exogenous gene encoding a cytochrome P450 polypeptide or a reductase polypeptide.3. The method of claim 1 , wherein the aromatic substrate comprises guaiacol claim 1 , anisole or guaethol.4. The method of claim 1 , wherein the material containing the aromatic substrate comprises products of lignin depolymerization.5. The method of claim 1 , wherein the material containing the aromatic substrate comprises a pyrolysis oil or bio-oil.6. The method of claim 1 , wherein at least one of the cytochrome P450 polypeptide or the reductase polypeptide is from a bacterium.7AmycolatopsisRhodococcus.. The method of claim 1 , wherein at least one of the cytochrome P450 polypeptide or the reductase polypeptide is from a bacterium from the genera or8. The method of claim 1 , wherein the cytochrome P450 polypeptide has an amino acid sequence at least 80% identical to SEQ ID No:2.9. The method of claim 1 , wherein the reductase polypeptide has an amino acid sequence at least 80% identical to SEQ ID NO:4.10. The method of claim 1 , wherein the dealkylation product is catechol or phenol.11. The method of claim 1 , further comprising isolating the dealkylation product.1220-. (canceled) This application claims ...

MICROORGANISMS AND METHODS FOR THE FERMENTATION OF CANNABINOIDS

Disclosed herein are microorganism and methods that can be used for the synthesis of cannabigerolic acid (CBGA) and cannabinoids. The methods disclosed can be used to produce CBGA, Δ-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), cannabichromenic acid (CBGA), Δ-tetrahydrocannabivarinic acid (THCVA), cannabidivarinic acid (CBDVA), cannabichromevarinic acid (CBCVA), Δ-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabichromene (CBC). Enzymes useful for the synthesis of CBGA and cannabinoids, include but are not limited to acyl activating enzyme (AAE1), polyketide synthase (PKS), olivetolic acid cyclase (OAC), prenyltransferase (PT), THCA synthase (THCAS), CBDA synthase (CBDAS), CBC A synthase (CBCAS), HMG-Co reductase (HMG1), and/or famesyl pyrophosphate synthetase (ERG20). The microorganisms can also have one or more genes disrupted, such as gene that that controls beta oxidation of long chain fatty acids. 1. A genetically modified microorganism comprising at least three polynucleotides that encode for:a) an amino acid sequence that is substantially identical, at least about 75% identical, at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 91% identical, at least about 92% identical, at least about 93% identical, at least about 94% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, or at least about 99% identical to SEQ ID NO: 27;b) an amino acid sequence that is substantially identical, at least about 75% identical, at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 91% identical, at least about 92% identical, at least about 93% identical, at least about 94% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, or at least about 99% identical to SEQ ID NO: 32; orc) combinations thereof.2 ...

KETOREDUCTASE POLYPEPTIDES FOR THE REDUCTION OF ACETOPHENONES

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. 1Lactobacillus. An engineered ketoreductase polypeptide derived from a wild-type ketoreductase having ketoreductase activity , wherein said engineered ketoreductase polypeptide is capable of converting in less than 24 hours at least 95% of the 2′ ,6′-dichloro-3′-fluoroacetophenone substrate to (S)-1-(2 ,6-dichloro-3-fluorophenyl) ethanol when carried out at an amount of less than 1% by weight with respect to the amount of the 2′ ,6′-dichloro-3′-fluoroacetophenone substrate , and wherein said engineered ketoreductase polypeptide comprises an amino acid sequence at least 90% identical to SEQ ID NO:2 , 4 , or 98 , and further wherein said amino acid sequence of said engineered ketoreductase polypeptide comprises at least one substitution at a position corresponding to a position in SEQ ID NO:2 , 4 , or 98 , selected from the following substitutions:the residue at the position corresponding to position 7 is an aromatic, non-polar, polar, constrained, or basic residue;the residue at the position corresponding to position 16 is a polar residue;the residue at the position corresponding to position 43 is a nonpolar or polar residue;the residue at the position corresponding to position 60 is an aromatic or non-polar, or aliphatic residue;the residue at the position corresponding to position 94 is a cysteine, non-polar or an aliphatic residue;the residue at the position corresponding to position 95 is a non-polar or aliphatic residue;the residue at the position corresponding to position 96 is a polar or acidic residue;the residue at the position corresponding to ...

METHOD FOR PRODUCING HIGH VALUE-ADDED COMPOUNDS FROM POLYETHYLENE TEREPHTHALATE

The present invention pertains to a method for producing high value-added compounds from polyethylene terephthalate. More specifically, the present invention demonstrates that a monomeric terephthalic acid obtained from the chemical hydrolysis of polyethylene terephthalate can be converted to high value-added aromatic compounds and aromatic-derived compounds, and ethylene glycol, which is another monomer of polyethylene terephthalate, can be converted to glycolic acid, which is a cosmetic material. The present invention is characterized by recycling polyethylene terephthalate waste into high value-added compounds. 1. A method of producing a high value-added compound from polyethylene terephthalate , comprising:producing terephthalic acid and ethylene glycol through hydrolysis of polyethylene terephthalate; andproducing one or more compounds selected from the group consisting of gallic acid, pyrogallol, catechol, muconic acid, and vanillic acid through bioconversion of the terephthalic acid in the presence of a biocatalyst, wherein protocatechuic acid is an intermediate produced by the bioconversion, orproducing glycolic acid through fermentation of the ethylene glycol.2. The method of claim 1 , wherein the hydrolysis of polyethylene terephthalate is performed by applying microwaves.3. The method of claim 1 , wherein the bioconversion of terephthalic acid to protocatechuic acid is performed using a microbe expressing terephthalic acid 1 claim 1 ,2-dioxygenase and 1 claim 1 ,2-dihydroxy-3 claim 1 ,5-cyclohexadiene-1 claim 1 ,4-dicarboxylate dehydrogenase as a biocatalyst.4. The method of claim 1 , wherein the bioconversion of terephthalic acid to gallic acid is performed using a microbe expressing terephthalic acid 1 claim 1 ,2-dioxygenase claim 1 , 1 claim 1 ,2-dihydroxy-3 claim 1 ,5-cyclohexadiene-1 claim 1 ,4-dicarboxylate dehydrogenase claim 1 , and p-hydroxybenzoate hydroxylase as a biocatalyst claim 1 , or using a combination of a microbe expressing terephthalic ...

OLIVETOL SYNTHASE VARIANTS AND METHODS FOR PRODUCTION OF OLIVETOLIC ACID AND ITS ANALOG COMPOUNDS

Described herein are non-natural olivetol synthase (OLS) variants, nucleic acids, engineered cells, method s for preparing cannabinoids, and compositions thereof. The non-natural olivetol OLS variants form desired cannabinoid precursor and products at increased rates, have higher affinity for pathway substrates, and/or byproducts are formed in lower amounts in their presence, as compared to wild type OLS. The OLS variants can be used to form linear polyketides, and can be expressed in an engineered cell having a pathway to form cannabinoids, which include CBGA, its analogs and derivatives. CBGA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions. 1. A non-natural olivetol synthase (OLS) comprising at least one amino acid variation as compared to a wild type olivetol synthase , wherein the non-natural olivetol synthase:(a) forms olivetolic acid or olivetol from malonyl-CoA and hexanoyl-CoA at a greater rate as compared to the wild type olivetol synthase;(b) has a higher affinity for hexanoyl-CoA and/or other acyl-CoA substrates as compared to the wild type olivetol synthase;(c) forms olivetolic acid analogs, olivetol analogs, variants thereof, or combinations thereof from malonyl-CoA and other acyl-CoAs at a greater rate as compared to the wild type olivetol synthase;(d) is characterized by a lower amount of one or more pyrone-based compounds being formed in the presence of the non-natural olivetol synthase (OLS) as compared to the wild type olivetol synthas, or(e) any combination of (a), (b), (c) or (d),wherein olivetolic acid or olivetol, analogs thereof, variants thereof, or acid derivatives of a polyketide are formed in the presence of olivetolic acid cyclase (OAC) not rate limited by amount or activity.2. The non-natural olivetol synthase of claim 1 , whereinthe pyrone-based hydrolysis compound is selected from pentyl diacetic acid lactone (PDAL), hexanoyl triacetic acid lactone (HTAL), and lactone analogs and ...

KETOREDUCTASE POLYPEPTIDES FOR THE REDUCTION OF ACETOPHENONES

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. 1. A recombinant ketoreductase polypeptide capable of stereoselectively reducing acetophenone to (S)-1-phenethanol , which comprises an amino acid sequence that is at least 95% identical to the reference sequence of SEQ ID NO: 119.2. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of stereoselectively reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol with a percent stereomeric excess of at least 99%.3. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate to the product at a rate greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.4. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (5)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol at a rate that is at least 450% greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.5. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (5)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol at a rate that is at least 1500% greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.6. The recombinant ketoreductase polypeptide of ...

AROMATIC PRENYLTRANSFERASE FROM CANNABIS

Nucleic acid molecules from has been isolated and characterized and encode polypeptides having aromatic prenyltransferase activity. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds. 2. (canceled)4. (canceled)5. (canceled)6. A vector claim 1 , constructs or expression system comprising the nucleic acid molecule as defined in .7. A process of transferring a prenyl group comprising: reacting a prenyl group acceptor molecule with a prenyl group donor molecule in presence of an aromatic prenyltransferase as defined in claim 3 , under suitable conditions for transferring the prenyl group from the prenyl group donor molecule to the prenyl group acceptor molecule.8. A method of altering levels of cannabinoid compounds in an organism claim 1 , cell or tissue comprising expressing a nucleic acid molecule as defined in claim 1 , or a part thereof claim 1 , to silence an aromatic prenyltransferase gene in the organism claim 1 , cell or tissue claim 1 , in comparison to a similar variety of organism claim 1 , cell or tissue grown under similar conditions but without the use of the nucleic acid molecule for silencing.9. An in vitro method of altering levels of cannabinoid compounds in a cell comprising expressing or over-expressing a nucleic acid molecule as defined in in the cell claim 1 , in comparison to a similar variety of cell grown under similar conditions but without the expressing or over-expressing of the nucleic acid molecule.10. An in vitro method of increasing levels of cannabinoid compounds in a cell comprising expressing or over-expressing a nucleic acid molecule encoding a polypeptide as defined in in the cell claim 3 , in comparison to a similar variety of cell grown under similar conditions but without the expressing or over-expressing of the nucleic acid molecule.11. The method of claim 9 , wherein the cell is or is from a microorganism ...

Optimized Microbial Cells for Production of Melatonin and Other Compounds

Described herein are recombinant microbial host cells comprising biosynthetic pathways and their use in producing oxidation products and downstream products, e.g., melatonin and related compounds, as well as enzyme variants, nucleic acids, vectors and methods useful for preparing and using such cells. In specific aspects, the present invention relates to monooxygenases, e.g., amino acid hydroxylases, with a modified cofactor-dependency, and to enzyme variants and microbial cells providing for an improved supply of cofactors. 1E. coliE. coli. A variant of GTP cyclohydrolase I (GCH1) having at least about 80% sequence identity to native GCH1 (SEQ ID NO:16) and comprising one or more mutations , wherein ,{'i': E. coli', 'E. coli, 'in an cell comprising a pterin-4α-carbinolamine dehydratase (PCD) and at least one of a tryptophan hydroxylase (TPH), a tyrosine hydroxylase (TH) and a phenylalanine hydroxylase (PheH), the variant provides for an increased hydroxylation activity of at least one of the TPH, TH and PheH as compared to native GCH1, and'}the mutation is not T198P.2. The variant of claim 1 , wherein at least one of the one or more mutations is in an amino acid residue in a segment selected from D97-E112 claim 1 , K121-D130 claim 1 , N170-H180 claim 1 , S193-L200 and S207-N222.3. The variant of claim 1 , wherein at least one of the one or more mutations is in an amino acid residue selected from the group consisting of D97 claim 1 , M99 claim 1 , T101 claim 1 , V102 claim 1 , A125 claim 1 , K129 claim 1 , N170 claim 1 , V179 claim 1 , T196 claim 1 , T198 claim 1 , S199 claim 1 , L200 claim 1 , S207 claim 1 , H212 claim 1 , E213 claim 1 , F214 claim 1 , L215 and H221.4. The variant of claim 1 , wherein the variant(a) comprises a mutation selected from D97V, D97L, D97A, D97T, M99C, M99T, M99V, M99L, M99I, T101I, T101V, T101L, V102M, N170K, N170D, N170L, V179A, V179M, T196I, T196V, T196L, T198I, T198V, T198S, T198L, S199Y, S199F, L200P, L200C, L200S, L200A, S207R, ...

METHOD FOR THE BIOCATALYTIC CYCLIZATION OF TERPENES AND CYCLASE MUTANTS EMPLOYABLE THEREIN

The present invention relates to novel mutants with cyclase activity and use thereof in a method for biocatalytic cyclization of terpenes, such as in particular for the production of isopulegol by cyclization of citronellal; a method for the preparation of menthol and methods for the biocatalytic conversion of further compounds with structural motifs similar to terpene. 4. The method of claim 2 , in which the compound of formula IV is selected from citronellal claim 2 , citral claim 2 , farnesol claim 2 , homofarnesol claim 2 , homofarnesol derivatives claim 2 , homofarnesylic acid claim 2 , geranylacetone claim 2 , melonal claim 2 , nonadienal claim 2 , and trimethyldecatetraene.5. The method of for the cyclization of terpenes and/or terpenoids claim 2 , and for the conversion of compounds of the general formula IV claim 2 , comprising utilizing:(a) said enzyme mutant;(b) a nucleic acid coding for said enzyme mutant;(c) an expression construct comprising said nucleic acid;(d) a recombinant vector comprising, under the control of at least one regulatory element, at least one of the nucleic acid of (b) or at least one of the expression construct of (c); or(e) a recombinant microorganism comprising the nucleic acid of (b), the expression construct of (c), or the recombinant vector of (d).6. The method of for the conversion of citronellal to isopulegol claim 5 , or for the conversion of squalene to hopene.7. The method of claim 1 , wherein up to 5% of the amino acid residues in said enzyme mutant are altered relative to SEQ ID NO: 209 by deletion claim 1 , insertion claim 1 , substitution claim 1 , addition claim 1 , inversion claim 1 , or a combination thereof.8. The method of claim 1 , wherein said mutation at the position corresponding to position F445 of the amino acid sequence of SEQ ID NO: 209 is a substitution selected from the group consisting of F445N claim 1 , F445Q claim 1 , F445L claim 1 , F445M claim 1 , F445E claim 1 , F445G claim 1 , F445S claim 1 , ...

Production of Stilbenoids

A method for the production of a stilbenoid, such as resveratrol or pinosylvin, by fermenting plant material such a grape must using a yeast having a metabolic pathway producing said stilbenoid, separating a solids waste material from said fermentation and extracting said stilbenoid.

DYNAMIC KNOCKDOWN OF CENTRAL METABOLISM FOR REDIRECTING GLUCOSE-6-PHOSPHATE FLUXES

Described herein are methods for dynamic redirection of metabolic flux in a cell from central metabolism towards production of heterologous products. 1. A method of redirecting flux of glucose-6-phosphate in a recombinant cell , the method comprising regulating activity of a phosphofructokinase-1 (pfk-1) in the recombinant cell.2. The method of claim 1 , further comprising expressing in the cell a heterologous pathway that can utilize a glycolytic intermediate.3. The method of or claim 1 , wherein the glycolytic intermediate is glucose-6-phosphate.4. The method of any one of - claim 1 , wherein the heterologous pathway comprises expressing a myo-inositol-1-phosphate synthase.5. The method of any one of - claim 1 , further comprising reducing expression of a glucose-6-phosphate dehydrogenase (zwf).6. The method of any one of - claim 1 , wherein the cell does not express glucose-6-phosphate dehydrogenase.7. The method of any one of - claim 1 , wherein regulating activity of the phosphofructokinase-1 protein comprises reducing the amount of phosphofructokinase-1 protein in the cell.8. The method of claim 7 , wherein the amount of phosphofructokinase-1 protein is reduced by at least 50%.9. The method of claim 7 , wherein the amount of phosphofructokinase-1 protein is reduced by at least 75%.10. The method of claim 7 , wherein the amount of phosphofructokinase-1 protein is reduced by at least 90%.11. The method of any one of - claim 7 , wherein reducing the amount of phosphofructokinase-1 protein comprises degrading the phosphofructokinase-1 protein.12. The method of any one of - claim 7 , wherein reducing the amount of phosphofructokinase-1 protein comprises targeting the phosphofructokinase-1 protein for degradation by a protease.13. The method of any one of - claim 7 , wherein the phosphofructokinase-1 protein is fused to a peptide tag.14. The method of claim 13 , wherein the peptide tag is an SsrA tag.15. The method of any one of - claim 13 , further comprising ...

USE OF TYPE III POLYKETIDE SYNTHASES AS PHLOROGLUCINOL SYNTHASES

Methods for producing phloroglucinol as well as methods of using type III polyketide synthases as phloroglucinol synthases, in particular the type III polyketide synthases of algae, such as eukaryotic ochrophyte algae, are described herein. In addition, polypeptides that have phloroglucinol synthase activity, the isolated nucleic acid molecules encoding these type III polyketide synthases, and the vectors and the host cells comprising such nucleic acid molecules are also described. 122.-. (canceled)23. A method comprising:using at least one polypeptide selected from type III polyketide synthases of algae as phloroglucinol synthase,{'i': 'Ectocarpus siliculosus', 'wherein the type III polyketide synthases of algae exclude type III polyketide synthases of selected from the group consisting of PKS1.Es, PKS2.Es and PKS3.Es.'}24. The method according to claim 23 , wherein said polypeptide is polyketide synthases of eukaryotic algae.25. The method according to claim 23 , wherein said polypeptide is type III polyketide synthases of ochrophyte algae.26. The method according to claim 23 , wherein said polypeptide comprises at least one amino acid sequence having at least 80% identity with a sequence selected from the group consisting of SEQ ID NO.: 1 and SEQ ID NO.: 3.27AureococcusSargassum. The method according to claim 25 , wherein said ochrophyte algae are selected from the group consisting of sp. and sp.28. The method according to claim 23 , wherein said polypeptide comprises at least one amino acid sequence having at least 85% identity with a sequence selected from the group consisting of SEQ ID NO.: 1 and SEQ ID NO.: 3.29. An isolated polypeptide with phloroglucinol synthase activity comprising at least one amino acid sequence having at least 80% identity with the sequence SEQ ID NO.: 3.30. The isolated polypeptide according to claim 29 , wherein the isolated polypeptide is recombinant.31. An isolated nucleic acid molecule encoding the isolated polypeptide according to ...

Recombinant production systems for aromatic molecules

The invention relates to the production of aromatic molecules in prokaryotic and eukaryotic hosts such as E. coli , yeasts, filamentous fungi, algae, microalgae, other plant cells.

Genetically Engineered Strain

The present disclosure discloses a genetically engineered strain, belonging to the technical field of bioengineering. L-amino acid oxidase genes, α-keto acid decarboxylase genes, alcohol dehydrogenase genes, and enzyme genes capable of reducing NAD(P) to NAD(P)H are introduced into the genetically engineered strain of the present disclosure. The present disclosure further discloses a construction method and application of a recombinant genetically engineered strain. When being applied to the biosynthesis of phenylethanoids, the method of the present disclosure has the characteristics of simple operation, low cost, and high synthesis efficiency and optical purity of the product, and has good industrialization prospects. 1Escherichia coli. A recombinant strain , which is capable of producing phenylethanoids at low cost , and capable of simultaneously expressing four enzymes which are L-amino acid oxidase , α-keto acid decarboxylase , alcohol dehydrogenase , and an enzyme capable of reducing NAD(P) to NAD(P)H.2Escherichia coliProteus mirabilisCosenzaea myxofaciens. The recombinant strain according to claim 1 , characterized in that the L-amino acid oxidase is from ATCC 29906 or ATCC 19692.3Escherichia coliProteus mirabilisLactococcus lactis. The recombinant strain according to claim 1 , characterized in that the α-keto acid decarboxylase is from ATCC 29906 or ATCC 19435.4Escherichia coliEscherichia coli. The recombinant strain according to claim 1 , characterized in that the alcohol dehydrogenase is from BL21(DE3).5Escherichia coli. The recombinant strain according to claim 1 , characterized in that the enzyme capable of reducing NAD(P) to NAD(P)H is formate dehydrogenase claim 1 , glucose dehydrogenase or phosphite dehydrogenase.6Escherichia coliKomagataella phaffiiBacillus subtilisPseudomonas abietaniphila. The recombinant strain according to claim 5 , characterized in that the enzyme capable of reducing NAD(P) to NAD(P)H is formate dehydrogenase from ATCC 76273 ...

ADDITIVE FOR RESINS

Provided are an abrasion resistance improver for inorganic filler-containing rubber composition capable of giving high abrasion resistance to a rubber composition containing an inorganic filler, a rubber composition containing the abrasion resistance improver for rubber composition, a tire using the rubber composition, and a method for producing the abrasion resistance improver for rubber composition. The present invention are concerned with [1] an abrasion resistance improver for inorganic filler-containing rubber composition, including, as an active ingredient, lignin having an aldehyde yield rate by alkaline nitrobenzene oxidation of 12% by mass or more; [2] a rubber composition containing the abrasion resistance improver as set forth above in [1], a rubber, and an inorganic filler; [3] a tire using the rubber composition as set forth above in [2]; [4] a method for producing an abrasion resistance improver for inorganic filler-containing rubber composition, including, as an active ingredient, lignin having an aldehyde yield rate by alkaline nitrobenzene oxidation of 12% by mass or more, the method including steps (A-1) to (A-3); and [5] a method for producing an abrasion resistance improver for inorganic filler-containing rubber composition, including, as an active ingredient, lignin having an aldehyde yield rate by alkaline nitrobenzene oxidation of 12% by mass or more, the method including steps (B-1) and (B-2). 1. A rubber composition comprising lignin having an aldehyde yield rate by alkaline nitrobenzene oxidation of 12% by mass or more , a rubber , and an inorganic filler.2. The rubber composition according to claim 1 , wherein the lignin has a weight average molecular weight of 500 or more and 30 claim 1 ,000 or less.3. The rubber composition according to claim 1 , wherein the content of lignin is 0.5 parts by mass or more and 30 parts by mass or less based on 100 parts by mass of the rubber.4. The rubber composition according to claim 1 , wherein the ...

Biosynthesis of Phenylpropanoid and Dihydrophenylpropanoid Derivatives

Provided herein are methods and compositions for producing phenylpropanoid derivatives, such as chalcones and stilbenes, and dihydrophenylpropanoid derivatives, such as dihydrochalcones and dihydrostilbenes, in microorganisms. In particular, the disclosure provides recombinant microorganisms and methods of use thereof for producing phenylpropanoid derivative compounds and dihydrophenylpropanoid derivative compounds.

METABOLICALLY ENGINEERED CELLS FOR THE PRODUCTION OF RESVERATROL OR AN OLIGOMERIC OR GLYCOSIDICALLY-BOUND DERIVATIVE THEREOF

A recombinant micro-organism producing resveratrol by a pathway in which phenylalanine ammonia lyase (PAL) produces trans-cinnamic acid from phenylalanine, cinnamate 4-hydroxylase (C4H) produces 4-coumaric acid from said trans-cinnamic acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA, or in which L-phenylalanine- or tyrosine-ammonia lyase (PAL/TAL) produces 4-coumaric acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including , or 1. A method for producing resveratrol or an oligomeric or glycosidically-bound derivative thereof comprising:a) cultivating a recombinant micro-organism comprising an engineered operative metabolic pathway producing resveratrol or an oligomeric or glycosidically-bound derivative thereof in a culture media comprising a carbon substrate, wherein the culture media does not require an external source of coumaric acid; andb) recovering the resveratrol or the oligomeric or glycosidically-bound derivative thereof from the culture media.2. The method of claim 1 , wherein the micro-organism is fungi or bacteria.3. The method of claim 2 , wherein the micro-organism is a fungus claim 2 , wherein the fungus is yeast.4Saccharomyes.. The method of claim 3 , wherein the yeast is from the genus5. The method of claim 1 , wherein the carbon substrate is a fermentable carbon substrate.6. The method of claim 5 , wherein the fermentable carbon substrate is monosaccharides claim 5 , oligosaccharides or polysaccharides.7. The method of claim 5 , wherein the fermentable carbon substrate is glucose claim 5 , fructose claim 5 , galactose claim 5 , xylose claim 5 , arabinose claim 5 , mannose claim 5 , sucrose claim 5 , lactose claim 5 , erythrose claim 5 , threose or ...

KETOREDUCTASE POLYPEPTIDES FOR THE REDUCTION OF ACETOPHENONES

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. 1. A recombinant ketoreductase polypeptide capable of stereoselectively reducing acetophenone to (S)-1-phenethanol , which comprises an amino acid sequence that is at least 80% identical to the reference sequence of SEQ ID NO: 119.2. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of stereoselectively reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol with a percent stereomeric excess of at least 99%.3. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate to the product at a rate greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.4. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol at a rate that is at least 450% greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.5. The recombinant ketoreductase polypeptide of claim 1 , wherein the polypeptide is further capable of reducing the substrate 2′ claim 1 ,6′-dichloro-3′-fluoroacetophenone to the product (S)-1-(2 claim 1 ,6-dichloro-3-fluorophenyl) ethanol at a rate that is at least 1500% greater than the rate capable by the ketoreductase polypeptide having the sequence of SEQ ID NO:6.6. The recombinant ketoreductase polypeptide of ...

MUTANT ENZYMES

This invention relates to mutant enzymes with enhanced properties and processes for oxidation of organic compound substrates using such enzymes. 122-. (canceled)23. A mutant CYP102A (Cytochrome P450 family 102A sub-family member) enzyme , wherein said CYP102A enzyme comprises a fusion of a heme monooxygenase domain comprising a P450 fold to a reductase domain , and said mutant enzyme comprises substitutions at one or more amino acid residue positions in the polypeptide chain of the wild-type CYP102A enzyme corresponding to (i) amino acid residue positions 47 , 51 and 307 of SEQ ID NO:2 and/or (ii) amino acid residue positions 330 or 403 of SEQ ID NO: 2 , thereby enhancing the monooxygenase activity and/or altering the product selectivity of the mutant enzyme.24. The mutant CYP102A enzyme according to claim 23 , wherein the reductase domain is a diflavin reductase domain.25. The mutant CYP102A enzyme according to claim 23 , wherein the wild-type CYP102A enzyme is capable of oxidising medium chain linear or branched fatty acids at sub-terminal positions.26. The mutant CYP102A enzyme according to claim 23 , comprising substitutions at amino acid residue positions 47 claim 23 , 51 and 307 of SEQ ID NO:2.27. The mutant CYP102A enzyme according to claim 26 , comprising a mutation selected from the group consisting of Q307H claim 26 , Q307N claim 26 , Q3075 claim 26 , Q307T and Q307Y.28. The mutant CYP102A enzyme according to claim 26 , comprising the mutations R47L and/or Y51F.29. The mutant CYP102A enzyme according to claim 23 , additionally comprising substitutions at one or more amino acid residue positions in the polypeptide chain of the wild-type CYP102A enzyme corresponding to positions 82 claim 23 , 87 claim 23 , 171 claim 23 , 263 claim 23 , 267 claim 23 , 319 claim 23 , 328 and 401 of SEQ ID NO: 2.30. The mutant CYP102A enzyme according to claim 29 , comprising mutations selected from one or more of A82L claim 29 , F87A claim 29 , F87G claim 29 , F87L claim 29 , ...

Polypeptides Having Peroxygenase Activity and Polynucleotides Encoding Same

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 129-. (canceled)30. An isolated polypeptide having peroxygenase activity , selected from the group consisting of:(a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2;(b) a polypeptide encoded by a polynucleotide that hybridizes under medium stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii);(c) a polypeptide encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, or the cDNA sequence thereof;(d) a variant of the mature polypeptide of SEQ ID NO: 2 comprising a substitution, deletion, and/or insertion at one or more positions; and(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has peroxygenase activity.31. A composition comprising the polypeptide of .32. A detergent composition claim 30 , comprising a surfactant and a polypeptide according to .33. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted claim 30 , linear or branched claim 30 , aliphatic hydrocarbon having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3 claim 30 , comprising contacting the aliphatic hydrocarbon with hydrogen peroxide and a polypeptide according to .34. A method for hydroxylation in position 2 or 3 of the terminal end of an acyl group of a lipid claim 30 , comprising contacting the lipid with hydrogen peroxide and a polypeptide according to .35. A method for introducing a hydroxy or a keto group at the second or third carbon of at least two ends of a substituted or ...

CHEMICAL ENGINEERING PROCESSES AND APPARATUS FOR THE SYNTHESIS OF COMPOUNDS

The present invention provides methods for producing cannabinoids and cannabinoid analogs as well as a system for producing these compounds. The inventive method is directed to contacting a compound according to Formula I or Formula II with a cannabinoid synthase. 2. The method of claim 1 , wherein Ris selected from the group consisting of linear CH claim 1 , CH claim 1 , CH claim 1 , CH claim 1 , CH claim 1 , CHand CH.3. The method of claim 2 , wherein Ris a linear CHgroup.4. The method of claim 1 , further comprising conjugating the cannabinoid synthase to a solid support.5. The method of claim 1 , wherein the cannabinoid or cannabinoid analog is a single enantiomer.6. The method of claim 1 , wherein the enantiomeric purity of the cannabinoid or cannabinoid analog is at least 95%.7. The method of claim 6 , wherein the enantiomeric purity of the cannabinoid or cannabinoid analog is at least 99%.8. The method of claim 1 , wherein the cannabinoid synthase is tetrahydrocannabinolic acid synthase (THCA synthase).9. The method of claim 1 , wherein the cannabinoid synthase is a recombinant cannabinoid synthase claim 1 , and the method further comprises producing the recombinant cannabinoid synthase.10. The method of claim 9 , wherein the step of producing recombinant THCA synthase comprises overexpressing the THCA synthase.11Escherichia coli.. The method of claim 10 , wherein the THCA synthase is expressed in yeast or in12. The method of claim 1 , wherein the cannabinoid synthase is CBDA synthase.13. The method of claim 1 , wherein the solvent comprises one or more of dimethyl sulfoxide (DMSO) claim 1 , dimethyl formamide (DMF) claim 1 , iso-propoyl alcohol or cyclodextrin claim 1 , and wherein the amount of the solvent in the reaction mixture is between 5% and 30% (w/v).14. The method of claim 1 , wherein the solvent is DMSO.15. The method of claim 1 , further comprising isolating the cannabinoid or a cannabinoid analog.16. The method of claim 1 , further comprising ...

Microorganisms Engineered to Produce Phenol and Its Derivatives

Novel methods for the in vivo production of phenol from renewable substrates using a recombinant microorganism (FIG. 1 ). Additionally, methods for the in vivo production of catechol and cis,cis-muconic acid from renewable substrates using a recombinant microorganism are disclosed. A host cell expresses at least one gene encoding a polypeptide that possesses isochorismate synthase activity, at least one gene encoding a polypeptide that possesses isochorismate pyruvate lyase activity, and at least one gene encoding a polypeptide that possesses salicylic acid decarboxylase activity. In the case of catechol, the host cell must additionally express at least one gene encoding a polypeptide that possesses phenol 2-monooxy-genase activity. In the case of cis,cis-muconic acid, the host cell must additionally express at least one gene encoding a polypeptide that possesses phenol 2-monooxygenase activity and at least one gene encoding a polypeptide that possesses catechol-1,2-dioxygenase activity.

A METHOD FOR PRODUCING RESVERATROL

Recombinant hosts and methods for producing resveratrol in recombinant hosts are disclosed herein. 1. A recombinant host comprising:(a) a gene encoding a 3-Deoxy-D-arabinoheptulosonate 7-phosphate (DAHP) synthase polypeptide;(b) a gene encoding a chorismate mutase polypeptide; and(c) a gene encoding an acetyl-CoA carboxylase polypeptide;wherein at least one of the genes is a recombinant gene,wherein the host is capable of producing a stilbene.2. The host of claim 1 , wherein the DAHP synthase polypeptide comprises a tyrosine-sensitive 3-Deoxy-D-arabinoheptulosonate 7-phosphate synthase (ARO4) polypeptide having at least 65% identity to the amino acid sequence set forth in SEQ ID NO: 1.3. The host of claim 2 , wherein the DAHP synthase polypeptide comprises an ARO4 polypeptide having a mutation that is K229L.4. The host of claim 1 , wherein the DAHP synthase polypeptide is not feedback inhibited.5. The host of claim 1 , wherein the host further comprises disruption of a gene encoding a phenylalanine-inhibited 3-Deoxy-D-arabinoheptulosonate 7-phosphate synthase (ARO3) polypeptide claim 1 , whereby the host does not express ARO3.6. The host of claim 5 , wherein the ARO3 polypeptide has at least 75% identity to the amino acid sequence set forth in SEQ ID NO: 37. The host of claim 1 , wherein the chorismate mutase polypeptide comprises a chorismate mutase (ARO7) polypeptide having at least 50% identity to the amino acid sequence set forth in SEQ ID NO: 4.8. The host of claim 7 , wherein the chorismate mutase polypeptide comprises an ARO7 polypeptide having a mutation that is T226I or T226N.9. The host of claim 1 , wherein the chorismate mutase polypeptide is not feedback inhibited.10. The host of claim 1 , wherein the acetyl-CoA carboxylase polypeptide comprises an acetyl-CoA carboxylase alpha (ACC1) polypeptide having at least 70% identity to the amino acid sequence set forth in SEQ ID NO: 6.11. The host of claim 10 , wherein the acetyl-CoA carboxylase polypeptide ...

FUSION PROTEINS USEFUL LN THE PRODUCTION OF CANNABINOIDS

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 113-. (canceled)14. An isolated codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34: or an isolated codon-optimized nucleic acid sequence of nucleic acids 5841 to 8867 of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acids 5841 to 8867 of SEQ ID NO: 34.15. An isolated codon-optimized nucleic acid sequence of SEQ ID NO: 35 or a nucleic acid sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 35; or an isolated codon-optimized nucleic acid sequence of nucleic acids 5052 to 6671 of SEQ ID NO: 35 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acids 5052 to 6671 of SEQ ID NO: 35.16. An isolated ...

BIOSYNTHETIC METHODS FOR THE MODIFICATION OF CANNABINOIDS

Provided is a method of modifying a first cannabinoid into a second cannabinoid or a non-cannabinoid. The method comprises combining the first cannabinoid with an enzyme that can modify the first cannabinoid into the second cannabinoid or non-cannabinoid under conditions where the first cannabinoid is modified into the second cannabinoid or non-cannabinoid. Also provided is a non-naturally occurring enzyme that can modify a first cannabinoid into a second cannabinoid or a non-cannabinoid. A nucleic acid encoding that enzyme is additionally provided. Further provided is a non-naturally occurring nucleic acid that encodes an enzyme having the enzymatic activity of the above non-naturally occurring enzyme. An expression cassette comprising that nucleic acid is additionally provided. A cell comprising the above expression cassette is further provided. Also provided is a plant expression cassette comprising the above-identified nucleic acid. 176-. (canceled)77. A nucleic acid encoding an enzyme that can modify a first cannabinoid into a second cannabinoid or a non-cannabinoid , wherein the nucleic acid comprises any one of SEQ ID NOs:1-50.7888-. (canceled)89. An expression cassette comprising the nucleic acid of .9091-. (canceled)92. A cell comprising the expression cassette of claim 89 , capable of expressing the enzyme that can modify a first cannabinoid into a second cannabinoid or a non-cannabinoid.93. The cell of claim 92 , which is a bacterial cell.94. The cell of claim 92 , which is a yeast cell.95Saccharomyces, Candida, Pichia, Schizosaccharomyces, Scheffersomyces, Blakeslea, Rhodotorula,Yarrowia.. The yeast cell of claim 95 , which is a species of or96. The cell of claim 92 , further comprising a THC biosynthetic pathway that allows the yeast cell to produce the first cannabinoid.97. The cell of claim 96 , wherein the cell can synthesize the first cannabinoid from a non-cannabinoid.98. The cell of claim 96 , wherein the cell comprises a recombinant geranyl ...

CHEMICAL ENGINEERING PROCESSES AND APPARATUS FOR THE SYNTHESIS OF COMPOUNDS

The present invention provides methods for producing cannabinoids and cannabinoid analogs as well as a system for producing these compounds. The inventive method is directed to contacting a compound according to Formula I or Formula II with a cannabinoid synthase. 2. The method of claim 1 , wherein Ris selected from the group consisting of linear CH claim 1 , CH claim 1 , CH claim 1 , CH claim 1 , CH claim 1 , CHand CH.3. The method of claim 2 , wherein Ris a linear CHgroup.4. The method of claim 1 , further comprising contacting the compound of Formula II with a cannabinoid synthase to obtain a cannabinoid or cannabinoid analog; andwherein the cannabinoid synthase is selected from the group consisting of a cannabidiolic acid (CBDA) synthase and a tetrahydrocannabinolic acid (THCA) synthase.5. The method of claim 4 , wherein the cannabinoid synthase is CBDA synthase.6. The method of claim 4 , further comprising isolating the cannabinoid or cannabinoid analog.7. The method of claim 4 , further comprising decarboxylating the cannabinoid or cannabinoid analog.8. The method of claim 4 , wherein the cannabinoid or cannabinoid analog is a single enantiomer.9. The method of claim 4 , wherein the enantiomeric purity of the compound of Formula II is at least 95%.10. The method of claim 9 , wherein the enantiomeric purity of the cannabinoid or cannabinoid analog is at least 99%. This application is a Continuation of U.S. application Ser. No. 15/430,122, filed Feb. 10, 2017, incorporated herein by reference in its entirety, which is a Continuation of U.S. application Ser. No. 15/242,189, filed Aug. 19, 2016, incorporated herein by reference in its entirety, which is a Continuation of U.S. application Ser. No. 15/171,517, filed Jun. 2, 2016, now U.S. Pat. No. 9,526,715, incorporated herein by reference in its entirety, which is a Continuation of U.S. application Ser. No. 14/836,339, filed Aug. 26, 2015, now U.S. Pat. No. 9,359,625 issued Jun. 7, 2016, incorporated herein by ...

STEREOSPECIFIC CARBONYL REDUCTASES

Stereospecific carbonyl reductases SCR1, SCR2, and SCR3 are described herein as are nucleotide sequences that encode these reductases. These stereospecific carbonyl reductases have anti-Prelog selectivity and have specificities that are useful for fine biochemical synthesis. 1Candida parapsilosis. A method of reducing a carbonyl substrate , comprising contacting the substrate with a purified polypeptide , the sequence of which comprises an amino acid sequence that has at least 70% identity to a stereospecific carbonyl reductase , wherein the polypeptide has carbonyl reductase activity and does not comprise SEQ ID NO:1 , in conditions suitable to catalyze the reduction of the carbonyl substrate.2. The method of claim 1 , wherein the reduction takes place in the presence of a coenzyme.3. The method of claim 2 , wherein the coenzyme is NADPH.4. The method of claim 1 , wherein the carbonyl substrate comprises an α-ketoester claim 1 , a β-ketoester claim 1 , an aryl ketone or an aliphatic ketone.5. The method of claim 4 , wherein the carbonyl substrate comprises an α-ketoester.6. The method of claim 5 , wherein the α-ketoester is methyl pyruvate claim 5 , methyl phenylglyoxylate claim 5 , ethyl pyruvate or ethyl benzoylformate.7. The method of claim 4 , wherein the carbonyl substrate comprises a β-ketoester.8. The method of claim of claim 7 , wherein the β-ketoester is ethyl trifluoroacetoacetate claim 7 , methyl acetoacetate claim 7 , methyl 3-oxovalerate claim 7 , methyl 4-fluorobenzoylacetate claim 7 , ethyl acetoacetate claim 7 , ethyl 3-oxovalerate claim 7 , ethyl 4-chloroacetoacetate claim 7 , ethyl benzoylacetate claim 7 , or ethyl 3 claim 7 ,4-dimethoxybenzoylacetate.9. The method of claim 4 , wherein the carbonyl substrate comprises an aryl ketone.10. The method of claim 9 , wherein the aryl ketone is 2-hydroxyacetophenone claim 9 , or a derivative thereof.11. The method of claim 10 , wherein the aryl ketone is 2′-chloro-2-hydroxyacetophenone claim 10 , 3′- ...

BIOSYNTHESIS OF PHENYLPROPANOIDS AND PHENYLPROPANOID DERIVATIVES

Provided herein are recombinant hosts and methods for producing phenylpropanoid and phenylpropanoid derivative compounds. It was found that tyrosine ammonia lyase from A449 provides improved coumaric acid production. 1. A recombinant host comprising a recombinant gene encoding a tyrosine ammonia lyase (TAL) polypeptide , wherein the host is capable of producing a phenylpropanoid or a phenylpropanoid derivative compound , and wherein the TAL polypeptide uses tyrosine as a preferred substrate.2. The host of claim 1 , wherein the gene encoding a TAL polypeptide encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:31.3. The host of claim 1 , wherein the gene encoding a TAL polypeptide encodes a polypeptide having at least 65% identity to the amino acid sequence set forth in SEQ ID NO:31.4. The host of claim 1 , wherein the gene encoding the TAL polypeptide is overexpressed.5. The host of claim 1 , wherein the recombinant host is capable of producing an increased yield of a phenylpropanoid or a phenylpropanoid derivative compound claim 1 , as compared to a recombinant host not comprising the TAL gene.6. The host of claim 1 , further comprising a recombinant gene encoding:(a) a stilbene synthase (STS) polypeptide; or(b) a chalcone synthase (CHS) polypeptide.7. The host of claim 1 , further comprising one or more of:(a) a gene encoding a L-phenylalanine ammonia lyase (PAL) polypeptide;(b) a gene encoding a cinnamate-4-hydroxylase (C4H) polypeptide;(c) a gene encoding a NADPH:cytochrome P450 reductase polypeptide;(d) a gene encoding a 4-coumarate-CoA ligase (4CL) polypeptide; or(e) a gene encoding a chalcone isomerase (CHI) polypeptidewherein at least one of the genes is a recombinant gene.8. The host of claim 1 , wherein the phenylpropanoid compound is coumaric acid.9. The host of claim 1 , wherein the phenylpropanoid derivative compound is a stilbenoid compound or a chalcone compound.10. The host of claim 9 , wherein the stilbene is resveratrol or a ...

BIOSYNTHESIS OF CANNABINOIDS AND CANNABINOID PRECURSORS

Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro. 1. A host cell that comprises a heterologous polynucleotide encoding a polyketide synthase (PKS) , wherein the PKS comprises an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 7 , 15 , 145 , and 714 , or wherein the PKS comprises a conservatively substituted version of any one of SEQ ID NOs: 7 , 15 , 145 , and 714 , and wherein the host cell further comprises one or more heterologous polynucleotides encoding one or more of: a polyketide cyclase (PKC) , a prenyltransferase (PT) , and/or a terminal synthase (TS).2. The host cell of claim 1 , wherein relative to the sequence of SEQ ID NO: 7 claim 1 , the PKS comprises an amino acid substitution at a residue corresponding to position 28 claim 1 , 34 claim 1 , 50 claim 1 , 70 claim 1 , 71 claim 1 , 76 claim 1 , 88 claim 1 , 100 claim 1 , 151 claim 1 , 203 claim 1 , 219 claim 1 , 285 claim 1 , 359 claim 1 , and/or 385 in SEQ ID NO: 7.3. The host cell of claim 1 , wherein the PKS comprises:a) the amino acid P at a residue corresponding to position 28 in SEQ ID NO: 7;b) the amino acid Q at a residue corresponding to position 34 in SEQ ID NO: 7;c) the amino acid N at a residue corresponding to position 50 in SEQ ID NO: 7;d) the amino acid M at a residue corresponding to position 70 in SEQ ID NO: 7;e) the amino acid Y at a residue corresponding to position 71 in SEQ ID NO: 7;f) the amino acid I at a residue corresponding to position 76 in SEQ ID NO: 7;g) the amino acid A at a residue corresponding to position 88 in SEQ ID NO: 7;h) the amino acid P or T at a residue corresponding to position 100 in SEQ ID NO: 7;i) the amino acid P at a residue corresponding to position 151 in SEQ ID NO: 7;j) the amino acid K at a residue corresponding to position 203 in SEQ ID NO: 7;k) the amino acid C at a residue corresponding to position 219 in SEQ ID NO: 7;l) the amino acid A ...

Method for preparing vanillin by fermentation with eugenol as substrate

The present disclosure discloses a method for preparing vanillin by fermentation with eugenol as a substrate, preparing vanillin by a mixed fermentation of Penicillium simplicissimum OMK-68 and Bacillus sp. OMK-69, its potency reached 35 g/L, the mass conversion rate of eugenol reaches 83.3%. Penicillium simplicissimum OMK-68 in the present disclosure can use eugenol as a substrate for fermentation to prepare coniferyl alcohol, and its fermentation effect and substrate utilization rate are far better than wild-type Penicillium simplicissimum. The Bacillus sp. OMK-69 in the present disclosure can use coniferyl alcohol as a substrate for fermentation to prepare vanillin, and its fermentation effect and substrate utilization rate are far better than wild-type Bacillus sp.

Method for High-efficiency Production of Pinoresinol Using an H2O2 Auto-scavenging Cascade

The present invention provides a method for high-efficiency production of pinoresinol by use of an HOauto-scavenging enzymatic cascade. It uses eugenol as the substrate, which is relatively inexpensive and is industrially available. It uses an enzymatic cascade to remove HOproduced in the process of pinoresinol synthesis, thereby reducing its inhibitory effect on the enzyme activity. In addition, the present invention uses whole cells as a catalyst, which can continuously regenerate cofactors needed by the enzyme, thus eliminating the need for exogenous addition of expensive cofactors during the reaction. The yield of the present invention can reach 7.12 g/L and the conversion rate is 61.55%. 1Escherichia coliEscherichia coli. A method for high-efficiency production of pinoresinol using an HOauto-scavenging enzymatic cascade , wherein eugenol is used as a substrate and the whole cell of recombinant is used as a catalyst to convert eugenol to pinoresinol , wherein the recombinant over-expresses vanillyl alcohol oxidase and peroxidase or a fusion protein of the two enzymes.2. The method of claim 1 , comprising the steps of:{'i': 'Escherichia coli', 'a) culturing the recombinant and inducing over-expression of vanillyl alcohol oxidase and peroxidase or a fusion protein of the two enzymes;'}{'i': 'Escherichia coli', 'b) incubating eugenol in the culture medium of the recombinant ; and'}c) converting eugenol to pinoresinol by an enzymatic cascade with vanillyl alcohol oxidase and peroxidase.3Penicillium simplicissimumEscherichia coli. The method of claim 1 , wherein the vanillyl alcohol oxidase is from (PsVAO) and the peroxidase is from BL21 (DE3) (Prx02).4. The method of claim 3 , wherein the amino acid sequence of PsVAO is SEQ ID NO:1 and the amino acid sequence of Prx02 is SEQ ID NO:3.5Escherichia coli. The method of claim 2 , wherein the culture medium for converting eugenol to pinoresinol comprises 10-200 mM PBS claim 2 , the recombinant cell with a concentration of ...

GENES AND PROTEINS FOR AROMATIC POLYKETIDE SYNTHESIS

Nucleic acid molecules encoding polypeptides having polyketide synthase activity have been identified and characterized. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds in organisms. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds. 127-. (canceled)28. A process of synthesizing a naturally-occurring cannabinoid compound or a non-naturally occurring analog of a cannabinoid compound in a microorganism comprising expressing a nucleic molecule that encodes a polypeptide with at least 95% sequence identity to SEQ ID NO: 2 or that is a conservatively substituted amino acid sequence of the sequence set forth in SEQ ID NO: 2 and having polyketide cyclase activity , in the microorganism in the presence of a type III polyketide synthase enzyme , an alkanoyl CoA and malonyl CoA.29. The process of claim 28 , wherein the microorganism is a yeast or a bacteria.30Saccharomyces cerevisiaeE. coli.. The process of claim 28 , wherein the microorganism is yeast or31. The process of claim 28 , wherein the nucleic acid molecule is expressed or over-expressed in combination with expression or over-expression of one or more other nucleic acids that encode one or more enzymes in a cannabinoid biosynthetic pathway.32. The process of claim 31 , wherein the one or more enzymes in a cannabinoid biosynthetic pathway is one or more of an acyl CoA synthetase claim 31 , a polyketide cyclase claim 31 , an aromatic prenyltransferase or a cannabinoid-forming oxidocylase.33. The process of claim 32 , wherein the one or more enzymes in a cannabinoid biosynthetic pathway is one or more of a hexanoyl CoA synthetase claim 32 , a geranylpyrophosphate:olivetolate geranyltransferase claim 32 , a Δ9-tetrahydrocannabinolic acid synthase claim 32 , a cannabidiolic acid synthase or a cannabichromenic acid synthase.34. The process of claim 28 , wherein the cannabinoid compound is one or more of cannabigerolic acid claim 28 , Δ9- ...

Molecular Cloning and Expression of cDNA Encoding O-methyltransferase Isolated from Mangifera Indica

The present invention relates to the cloning and expression of cDNA encoding O-methyl transferase for catalyzing the synthesis of vital volatile flavor compounds such as mesifuran and vanillin, thereby playing a significant role in flavor biochemistry of mango. The present invention also provide a process for the preparation of volatile compounds by mesifuran, ferulic acid, guaiacol and vanillin, specifically mesifuran and vanillin using cDNA sequence. 1Mangifera indica. A nucleotide SEQ ID No.2 encoding for O-methyltransferase protein of (MiOMTS).2Mangifera indica. The nucleotide as claimed in claim 1 , wherein the O-methyltransferase (Mi-OMTS) protein is having SEQ ID No.3.3Mangifera indica. A protein molecule having SEQ ID No. 3 encoding for O-methyltransferase (Mi-OMTS).4. A recombinant expression vector comprising the nucleotide SEQ ID No. 2 of .5. The recombinant expression vector as claimed in claim 4 , wherein the said recombinant expression vector is selected from pGEX-4T claim 4 , pGEX-2T claim 4 , pGEX-3X claim 4 , pGEX-5X and pGEX-6P.6. The recombinant expression vector as claimed in claim 5 , wherein the said recombinant expression vector is pGEX-4T.7Escherichia coli.. A host cell comprising the recombinant expression vector as claimed in claim 5 , wherein the said host cell is8Escherichia coli.. A host cell comprising the DNA molecule of the SEQ ID No. 2 of claim 1 , wherein the said host cell is9. A process for synthesis of volatile compounds said process comprising the steps of:(a) cloning Mi-OMTS nucleotide SEQ ID No. 2 in a recombinant expression vector;{'i': 'E. coli;', '(b) inserting the recombinant expression vector of step (a) in'}{'i': 'E. coli;', '(c) expressing Mi-OMTS nucleotide SEQ ID No. 2 to produce Mi-OMTS protein having SEQ ID No. 3 in'}{'i': 'E. coli', '(d) extracting the said Mi-OMTS protein of step (c) by cell lysis;'}(e) carrying out chromatography and filtration of the extracted protein of step (d) to obtain purified protein ...

APPARATUS AND METHODS FOR BIOSYNTHETIC PRODUCTION OF CANNABINOIDS

The present invention provides an apparatus and methods for producing tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA) and cannabichromenic acid (CBCA) in different ratios. The apparatus comprises: (i) a bioreactor comprising (a) an automated supply system configured to deliver a first automated supply of cannabigerolic acid (CBGA), a cannabinoid acid synthase, and a reaction mixture; and (b) a second automated system to cease the reaction; (ii) a controller configured to modify a property of the reaction mixture to produce the desired products; and (iii) an extractor configured to recover the tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA) or cannabidiolic acid (CBDA) and cannabichromenic acid. 16.-. (canceled)7. A system for producing cannabinoids wherein the system comprises:(i) a bioreactor comprising cells stably transformed with one or more cannabinoid acid synthase genes;(ii) a reaction mixture; and(iii) a controller configured to modify one or more reaction conditions to modulate the ratio of the cannabinoid products.81. The system of claim , wherein the one or more cannabinoid acid synthase genes comprises a tetrahydrocannabinolic acid (THCA) synthase gene.92. The system of claim , wherein the THCA synthase gene comprises a nucleic acid sequence of SEQ ID NO: 1 , SEQ ID NO: 3 , or a variant or fragment thereof.102. The system of claim , wherein the THCA synthase gene comprises a nucleic acid sequence that encodes a polypeptide of SEQ ID NO: 2 , SEQ ID NO: 4 , or a variant or fragment thereof.111. The system of claim , wherein the one or more cannabinoid acid synthase genes comprises a cannabidiolic acid (CBDA) synthase gene.125. The system of claim , wherein the CBDA synthase gene comprises a nucleic acid sequence of SEQ ID NO: 5 , SEQ ID NO: 7 , or a variant or fragment thereof.135. The system of claim , wherein the CBDA synthase gene comprises a nucleic acid sequence that encodes a polypeptide of SEQ ID NO: 6 , SEQ ID NO: ...

POLYPEPTIDES HAVING PEROXYGENASE ACTIVITY

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 143-. (canceled)44. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted , linear or branched , aliphatic hydrocarbon having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3 , comprising contacting the aliphatic hydrocarbon with hydrogen peroxide and a polypeptide having peroxygenase activity , wherein the polypeptide has at least 80% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.45. A method for hydroxylation in position 2 or 3 of the terminal end of an acyl group of a lipid , comprising contacting the lipid with hydrogen peroxide and a polypeptide having peroxygenase activity , wherein the polypeptide has at least 80% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.46. A method for introducing a hydroxy or a keto group at the second or third carbon of at least two ends of a substituted or unsubstituted , linear or branched , aliphatic hydrocarbon having at least five carbons and having at least one hydrogen attached to said second or third carbon , comprising contacting the aliphatic hydrocarbon with hydrogen peroxide and a polypeptide having peroxygenase activity , wherein the polypeptide has at least 80% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.47. The method of claim 44 , wherein the aliphatic hydrocarbon is an alkane.48. The method of claim 45 , wherein the aliphatic hydrocarbon is an alkane.49. The method of claim 46 , wherein the aliphatic hydrocarbon is an alkane.50. The method of claim 47 , wherein the alkane is pentane claim 47 , hexane claim 47 , heptane claim 47 , octane claim 47 , nonane claim 47 ...

ANTIMALARIAL AGENT, METHODS AND USES THEREOF

The present disclosure relates to compounds and compositions for the treatment of malarian diseases. The compounds comprise a class of halogenated alkyl-aromatic secondary metabolites, including hierridic C, or pharmaceutically acceptable salts, esters, solvates or prodrugs thereof. The compounds comprise a halogen; C-Calkyl, H, COCH3, COH, CO-alkyl, or CO-aryl; and a C-Calkyl. In at least one embodiment, the compound is 3-chloro-4,6-dimethoxy-2-pentadecylphenol. The present disclosure also relates to methods of obtain the compounds from a solution comprising a cyanobacterium. 2. The compound of claim 1 , wherein R3 is a C10-C30 alkyl.3. The compound of claim 1 , whereinR1 is selected from the group consisting of F, Cl, and Br;R2, R4 are independently selected from a group consisting of C1-C6 alkyl; andR3 is a C15-C21 alkyl.4. The compound of claim 1 , wherein R2 and R4 are equal.5. The compound of claim 1 , wherein the compound is 3-chloro-4 claim 1 ,6-dimethoxy-2-pentadecylphenol.6. (canceled)7. A pharmaceutical composition comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a compound as defined in in a therapeutically effective amount; and'}a pharmaceutically acceptable excipient.8. The pharmaceutical composition of claim 7 , wherein the pharmaceutically acceptable excipient is a carrier claim 7 , adjuvant claim 7 , excipient or a mixture thereof.9. The pharmaceutical composition of claim 7 , wherein the composition is suitable for topical claim 7 , oral claim 7 , parental claim 7 , or injectable administration.10. (canceled)11. A method for obtaining the compound of 3-chloro-4 claim 7 , 6-dimethoxy-2-pentadecylphenol comprising: isolating said compound from a solution comprising a cyanobacterium strain with accession number CCAP1436/1.12. (canceled)13. (canceled)14. (canceled) This application is a U.S. National Stage Application under 35 U.S.C. § 371 of International Patent Application No. PCT/IB2016/053823, filed Jun. 27, 2016, which claims priority ...

Polypeptides Having Peroxygenase Activity

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 143.-. (canceled)44. A nucleic acid construct or expression vector comprising a polynucleotide encoding polypeptide having peroxygenase activity , wherein the polypeptide has at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 2 and the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in an expression host.45. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide has at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 2.46. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide has at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2.47. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.48. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1 claim 44 , (ii) the cDNA sequence thereof claim 44 , or (iii) the full-length complement of (i) or (ii).49. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide is encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 claim 44 , or the cDNA sequence thereof.50. The nucleic acid construct or expression vector of claim 44 , wherein the polypeptide is a variant of the mature polypeptide of SEQ ID NO: 2 comprising a substitution claim 44 , deletion claim 44 , and/or ...

AGLYCONE PRODUCTION PROMOTER

A technique is provided for degrading a resveratrol glycoside, or degrading a resveratrol glycoside and an isoflavone glycoside, to promote the production of an aglycone(s), thereby enhancing the absorption thereof into a living body. A bacterium belonging to the genus is used as an active ingredient of an aglycone production promoter which is used for promoting the production of an aglycone(s) from a glycoside(s). The aglycone production promoter is incorporated into a pharmaceutical composition for preventing and/or treating a disease for which an aglycone(s) is/are effective, or into a food or beverage composition for producing an aglycone(s). 19-. (canceled)10. A method of producing an aglycone(s) , the method comprising the steps of:{'i': 'Bifidobacterium', 'A) culturing a bacterium belonging to the genus in the presence of a glycoside(s); and'}B) collecting the aglycone(s) produced in a culture obtained by culturing the bacterium;wherein the glycoside(s) comprise(s) at least a resveratrol glycoside.11BifidobacteriumBifidobacterium breve.. The method according to claim 10 , wherein the bacterium belonging to the genus is12. The method according to or claim 10 , wherein the glycoside(s) further comprise(s) an isoflavone glycoside.13Bifidobacterium breveBifidobacterium breveBifidobacterium breveBifidobacterium breve, Bifidobacterium breve. The method according to claim 11 , wherein the is selected from the group consisting of ATCC 15700 claim 11 , BCCM LMG 23729 claim 11 , FERM BP-11175NITE BP-02460 claim 11 , and combinations thereof.14BifidobacteriumBifidobacterium breve. A bacterium belonging to the genus claim 11 , which is NITE BP-02460.1516-. (canceled)17Bifidobacterium. A method of promoting the production of an aglycone(s) from a glycoside(s) claim 11 , the method comprising the step of culturing a bacterium belonging to the genus in the presence of the glycoside(s) claim 11 ,wherein the aglycone(s) comprise(s) at least resveratrol.1821-. (canceled) This ...

METHODS AND ORGANISM WITH INCREASED XYLOSE UPTAKE

Provided herein are novel xylose transporters and their variants, as well as nucleic acid encoding the novel xylose transporters and their variants. Provided herein are also non-naturally occurring microbial organisms having increased xylose uptake and increased production of bioderived compounds using xylose as a substrate, as well as methods to make and use these microbial organisms. 1Metschnikowia. A non-naturally occurring microbial organism comprising at least one exogenous nucleic acid encoding a xylose transporter , wherein said xylose transporter has an amino acid sequence that is at least 89% identical to a xylose transporter.2. The non-naturally occurring microbial organism of claim 1 , comprising exogenous nucleic acids encoding at least two claim 1 , at least three claim 1 , at least four claim 1 , at least five claim 1 , at least six claim 1 , or at least seven xylose transporters.3Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter has an amino acid sequence that is at least 90% claim 1 , at least 95% claim 1 , at least 98% claim 1 , or at least 99% claim 1 , identical to said xylose transporter.4Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is a xylose transporter.5Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is selected from the group consisting of Xyt1p claim 1 , Gxf1p claim 1 , ΔGxf1p claim 1 , Gxf2p/Gal2p claim 1 , Gxs1p/Hgt12p claim 1 , ΔGxs1p/ΔHgt12p claim 1 , Hxt5p claim 1 , Hxt2.6p claim 1 , Qup2p claim 1 , and Aps1p/Hgt19p.6MetschnikowiaMetschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is from a H0 sp.7Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-5 and 7-12.8. The non- ...

TRANSFORMANT OF CORYNEFORM BACTERIUM AND PRODUCTION METHOD FOR USEFUL COMPOUND USING SAME

Provided is a transformant of a microorganism that has improved catechol productivity. 1. A transformant of a coryneform bacterium that is obtained by introducing , into the coryneform bacterium as a host , at least one gene selected from the group consisting of:{'i': 'Lactobacillus rhamnosus;', '(1) a decarboxylase gene ubiD of'}{'i': Lactobacillus', 'Bacillus', 'Enterobacter', 'Escherichia', 'Paenibacillus', 'Citrobacter', 'Pantoea, '(2) an ortholog of the gene (1) in at least one of the genus , the genus , the genus , the genus , the genus , the genus , or the genus ; and'}(3) a gene in which an enzyme that has an amino acid sequence identity of 70% or more with an amino acid sequence of an enzyme encoded by the gene (1) or (2), and that has a decarboxylation activity, is encoded,wherein mutations are introduced into a catechol 1,2-dioxygenase gene catA, and a protocatechuic acid dehydrogenase gene pcaHG in the coryneform bacterium as a host; and functions of enzymes encoded by these two genes are degraded or lost.2. The transformant according to claim 1 ,wherein the transformant has a catechol producing ability.3. The transformant according to claim 1 ,wherein at least one of a gene that encodes an enzyme having 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase activity, or a gene that encodes an enzyme having 3-dehydroquinate synthase activity, is additionally introduce.4. The transformant according to claim 1 ,{'i': 'Corynebacterium glutamicum', 'wherein the coryneform bacterium as a host is R (FERM P-18976), ATCC13032, or ATCC13869.'}5Corynebacterium glutamicum. A transformant of CAT21 (Accession Number: NITE BP-02689).6. A method for producing catechol comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'causing the transformant according to to react in a reaction solution in which at least one of factors necessary for growth is removed, or in a reaction solution under reducing conditions; and'}collecting catechol in a reaction medium.7. The ...

CO-CULTURE BASED MODULAR ENGINEERING FOR THE BIOSYNTHESIS OF ISOPRENOIDS, AROMATICS AND AROMATIC-DERIVED COMPOUNDS

The invention relates to co-cultures and their use in the biosynthesis of functionalized taxanes, other isoprenoids, aromatics, and aromatic-derived compounds. 1. A synthetic cellular consortium , comprisinga first organism comprising a first part of a biosynthetic pathway that produces a first compound anda second organism comprising a second part of the biosynthetic pathway that is able to convert the first compound into a second compound; andoptionally comprising a third organism that converts the second compound into a third compound.2. The synthetic cellular consortium of claim 1 , wherein the first and/or second organism is{'i': Escherichia coli, Bacillus subtilis', 'Bacillus megaterium', 'E. coli, B. subtilis', 'B. megaterium, 'a bacterium, optionally wherein the bacterium is , or , optionally wherein the or is genetically engineered;'}{'i': Saccharomyces cerevisiae, Yarrowia lipolytica', 'Pichia pastoris', 'S. cerevisiae, Y. lipolytica', 'P. pastoris, 'a yeast, optionally wherein the yeast is , or , optionally wherein the , or is genetically engineered; or'}{'i': Taxus', 'Taxus', 'Taxus, 'a plant cell, optionally wherein the plant cell belongs to the genus , optionally wherein the cell is induced with methyl jasmonate, optionally wherein the cell is genetically engineered.'}34-. (canceled)5. The synthetic cellular consortium of claim 1 , wherein the first organism recombinantly expresses one or more enzymes of a biosynthetic pathway claim 1 , optionally the shikimate pathway or a secondary metabolite biosynthetic pathway claim 1 , optionally wherein the secondary metabolite biosynthetic pathway is an isoprenoid biosynthetic pathway claim 1 , optionally a 2-C-methyl-D-erythritol 4-phosphate/1-deoxy-D-xylulose 5-phosphate (MEP) pathway.68-. (canceled)9. The synthetic cellular consortium of claim 5 , wherein the first organism recombinantly expresses{'i': 'E. coli;', '(i) any of the genes dxs, idi, ispD, ispF of the MEP pathway, and/or any of the genes ispG and ...

METHODS AND ORGANISM WITH INCREASED XYLOSE UPTAKE

Provided herein are novel xylose transporters and their variants, as well as nucleic acid encoding the novel xylose transporters and their variants. Provided herein are also non-naturally occurring microbial organisms having increased xylose uptake and increased production of bioderived compounds using xylose as a substrate, as well as methods to make and use these microbial organisms. 1Metschnikowia. A non-naturally occurring microbial organism comprising at least one exogenous nucleic acid encoding a xylose transporter , wherein said xylose transporter has an amino acid sequence that is at least 89% identical to a xylose transporter.2. The non-naturally occurring microbial organism of claim 1 , comprising exogenous nucleic acids encoding at least two claim 1 , at least three claim 1 , at least four claim 1 , at least five claim 1 , at least six claim 1 , or at least seven xylose transporters.3Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter has an amino acid sequence that is at least 90% claim 1 , at least 95% claim 1 , at least 98% claim 1 , or at least 99% claim 1 , identical to said xylose transporter.4Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is a xylose transporter.5Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is selected from the group consisting of Xyt1p claim 1 , Gxf1p claim 1 , ΔGxf1p claim 1 , Gxf2p/Gal2p claim 1 , Gxs1p/Hgt12p claim 1 , ΔGxs1p/ΔHgt12p claim 1 , Hxt5p claim 1 , Hxt2.6p claim 1 , Qup2p claim 1 , and Aps1p/Hgt19p.6MetschnikowiaMetschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter is from a H0 sp.7Metschnikowia. The non-naturally occurring microbial organism of claim 1 , wherein said xylose transporter has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-5 and 7-12.8. The non- ...

Metschnikowia species for biosynthesis of compounds

Provided herein are Metschnikowia species that produce useful compounds from xylose when cultured, as well as methods to make and use these Metschnikowia species.

METHODS OF MAKING VANILLIN VIA THE MICROBIAL FERMENTATION OF FERULIC ACID FROM EUGENOL USING A PLANT DEHYDROGENASE

A bioconversion method of making vanillin including expressing VaoA gene in a mixture, expressing MtSAD1 gene in the mixture, feeding eugenol to the mixture, and converting ferulic acid to vanillin by incubating with a microbial sp. strain (Zhp06) and/or a recombinant strain. 1. A bioconversion method of making ferulic acid comprising the following steps:expressing a VaoA gene in a mixture;expressing a MtSAD1 gene in the mixture;feeding eugenol to the mixture; andcollecting ferulic acid.2coli.. The bioconversion method of making ferulic acid of claim 1 , wherein expressing MtSAD1 is based on amino acid sequence selected from the group consisting of: SEQ ID No. 1; an amino acid sequence with at least 95% identity to SEQ ID No. 1; an amino acid sequence with at least 90% identity to SEQ ID No. 1; and an amino acid sequence expressed from E claim 1 ,3E. coli.. The bioconversion method of making ferulic acid of claim 1 , wherein expressing VaoA is based on amino acid sequence selected from the group consisting of: SEQ ID No. 5; on an amino acid sequence with at least 95% identity to SEQ ID No. 5; an amino acid sequence with at least 90% identity to SEQ ID No. 5; and an amino acid sequence expressed from4. The bioconversion method of making ferulic acid of further comprising expressing an ADH gene claim 1 , wherein expressing ADH is based on amino acid sequence selected from the group consisting of: SEQ ID No. 3; on an amino acid sequence with at least 95% identity to SEQ ID No. 3; and an amino acid sequence with at least 90% identity to SEQ ID No. 3.5. The bioconversion method of making ferulic acid of claim 4 , wherein the method of expressing any of the genes is selected from the group consisting of: expressing the gene by in vitro translation; expressing the gene in a cellular system; and expressing the gene in bacteria or yeast.6. A composition comprising ferulic acid obtainable by the method of wherein the ferulic acid has a δC value range of −25 to −32 which ...

HOST CELLS AND METHODS FOR PRODUCING HYDROXYTYROSOL

The present invention provides for a composition comprising: (a) a first host cell capable of producing L-DOPA; and (b) a modified host cell is capable of converting L-DOPA into hydroxytyrosol (HTy); wherein any one or both of the first host cell and second host cell is a genetically modified host cell. 1. A composition comprising: (a) a first host cell capable of producing L-DOPA; and (b) a second host cell capable of converting L-DOPA into hydroxytyrosol (HTy); wherein any one or both of the first host cell and the second host cell is a genetically modified host cell.2. The composition of claim 1 , wherein the first host cell is a first genetically modified host cell claim 1 , and the second host cell is a second genetically modified host cell.3. The composition of claim 1 , wherein the first host cell comprises dihydropteridine reductase (DHPR) claim 1 , pterin-4-alpha-carbinolamine dehydratase (PCD) claim 1 , and tyrosine hydroxylase (TH) claim 1 , or any homologous enzyme thereof.4. The composition of claim 3 , wherein one or more of DHPR claim 3 , PCD claim 3 , and TH are heterologous to the first host cell.5. The composition of claim 1 , wherein the first host cell is engineered to overproduce tyrosine compared to a non-engineered cell claim 1 , and the first host cell comprises a tyrosine hydroxylase (TH) claim 1 , or a homologous enzyme thereof.6. The composition of claim 5 , wherein the first host cell overexpresses AroG claim 5 , or a homologous enzyme thereof claim 5 , and/or TyrA claim 5 , or a homologous enzyme thereof.7. The composition of claim 6 , wherein the first host cell further comprises one or more claim 6 , or all claim 6 , of the following enzymes claim 6 , or a corresponding homologous enzyme thereof claim 6 , for the synthesis of L-tyrosine: phosphoenolpyruvate synthase (PpsA) claim 6 , transketolase A (TktA) claim 6 , DAHP synthase (AroG) claim 6 , DHQ synthase (AroB) claim 6 , DHQ dehydratase (AroD) claim 6 , quinate/shikimate ...

BIOTECHNOLOGICAL METHODS FOR PROVIDING 3,4-DIHYDROXYPHENYL COMPOUNDS AND METHYLATED VARIANTS THEREOF

The present invention relates to genetically modified enzymes obtained by rational design of the active site binding pocket of the prototypic enzyme 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H) for hydroxylating a 4-hydroxyphenyl compound to yield a 3,4-dihydroxyphenyl compound and to biotechnological methods including in vivo and in vitro methods using said enzymes or catalytically active fragments thereof. Further provided is a method either using a suitable oxidase or hydroxylase further enabling the subsequent site specific methylation of the 3,4-dihydroxyphenyl compound in a coupled enzymatic reaction by providing a suitable O-methyltransferase. Finally, compositions obtainable by the aforementioned methods are disclosed. 1. A method for catalyzing the biotechnological conversion of at least one 4-hydroxyphenyl compound for producing at least one 3 ,4-dihydroxyphenol compound comprising:{'i': 'Escherichia coli', '(i) providing at least one amino acid sequence comprising a genetically modified derived 4-hydroxyphenylacetate 3-hydroxylase or a catalytically active fragment thereof, comprising at least one mutation in its active site;'}(ii) providing at least one 4-hydroxyphenyl compound;(iii) reacting the at least one 4-hydroxyphenyl compound and the at least one amino acid sequence comprising the 4-hydroxyphenylacetate 3-hydroxylase or the catalytically active fragment thereof under suitable reaction conditions for allowing the hydroxylation of the at least one 4-hydroxyphenyl compound by the at least one amino acid sequence comprising the 4-hydroxyphenylacetate 3-hydroxylase or the catalytically active fragment thereof to yield at least one 3,4-dihydroxyphenol compound; and(iv) optionally isolating and/or purifying the resulting at least one 3,4-dihydroxyphenol compound.2. The method according to claim 1 , wherein the method is performed as a whole-cell approach claim 1 , and wherein (i) is performed as follows:(a1) providing at least one recombinant host cell ...

PROCESS FOR PRODUCING 7-DEHYDROCHOLESTEROL AND VITAMIN D3

According to the present invention, there can be provided a process for producing 7-dehydrocholesterol (7DHC), comprising culturing, in a medium, a 7DHC-producing microorganism in which sterol 24-C-methyltransferase activity is reduced or lost as compared to a parent strain, allowing 7DHC to be produced and accumulated in the culture, and collecting the 7DHC from the culture; and a process for producing vitamin D3, comprising irradiating, with ultraviolet light, 7-dehydrocholesterol produced by the production process. 1. A process for producing 7-dehydrocholesterol (hereinafter , “7DHC”) , comprising:{'i': 'Labyrinthulea', 'culturing, in a medium, a 7DHC-producing microorganism in which sterol 24-C-methyltransferase activity (hereinafter, “SMT activity”) is reduced or lost as compared to a parent strain;'}allowing 7DHC to be produced and accumulated in the culture; andcollecting the 7DHC from the culture.2LabyrinthuleaLabyrinthulea. The production process according to claim 1 , wherein the microorganism in which the SMT activity is reduced or lost as compared to the parent strain is a microorganism in which the SMT activity is reduced or lost as compared to the parent strain through deletion claim 1 , substitution claim 1 , or addition of at least one base in a gene which is present in the chromosomal DNA of the parent strain and encodes a protein having SMT activity.3LabyrinthuleaLabyrinthulea. The production process according to claim 1 , wherein the microorganism in which the SMT activity is reduced or lost as compared to the parent strain is a microorganism in which the SMT activity is reduced or lost as compared to the parent strain claim 1 , being obtained by transforming the parent strain with a DNA encoding an RNA which has a sequence complementary to a messenger RNA transcribed from a gene present in the chromosomal DNA of the parent strain and encoding a protein having SMT activity claim 1 , and forms a conjugate with the messenger RNA.4. The production ...

Ketoreductase polypeptides for the reduction of acetophenones

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds.

Method for producing modified resveratrol

Methods for producing glycosylated and methylated resveratrol in a genetically engineered cell, by bioconversion, and in vitro are disclosed herein.