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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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02-08-2012 дата публикации

Novel Sesquiterpene Synthase Gene and Protein

Номер: US20120196340A1
Автор: Bryan Greenhagen, Joseph Chappell
Принадлежит: University of Kentucky Research Foundation

The invention relates to sesquiterpene synthases and methods for their production and use. Particularly, the invention provides nucleic acids comprising the nucleotide sequence of citrus valencene synthase (CVS) which codes for at least one CVS. The invention further provides nucleic acids comprising the nucleotide sequence coding for amino acid residues forming the tier 1 and tier 2 domains of CVS. The invention also provides for methods of making and using the nucleic acids and amino acids of the current invention.

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Номер записи: 1
25-10-2012 дата публикации

Methods of developing terpene synthase variants

Номер: US20120270260A1
Автор: Andrew MAIN, Lan Xu, Lishan Zhao, Patrick WESTFALL
Принадлежит: Amyris Inc

The present disclosure relates to methods of developing terpene synthase variants through engineered host cells. Particularly, the disclosure provides methods of developing terpene synthase variants with improved in vivo performance that are useful in the commercial production of terpene products. Further encompassed in the present disclosure are superior terpene synthase variants and host cells comprising such terpene synthase variants.

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Номер записи: 2
21-02-2013 дата публикации

Isoprene synthase variants for improved production of isoprene

Номер: US20130045891A1
Автор: Christopher L. Rife, David A. Estell, Derek H. Wells, Jeffrey V. Miller, Zachary Q. Beck
Принадлежит: DANISCO US INC

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved specific productivity. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in host cells.

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Номер записи: 3
25-04-2013 дата публикации

Method for producing patchoulol and 7-epi-alpha-selinene

Номер: US20130102038A1
Автор: Fabienne Deguerry, Michel Schalk
Принадлежит: FIRMENICH SA

A method of producing patchoulol and 7-epi-α-selinene by contacting at least one polypeptide with farnesyl phyrophosphate (fpp). The method may be carried out in vitro or in vivo to produce patchoulol and 7-epi-α-selinene, compounds which can be useful in the field of perfumery.

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Номер записи: 4
26-09-2013 дата публикации

Three-dimensional structure of isoprene synthase and its use thereof for generating variants

Номер: US20130252303A1
Автор: Christopher L. Rife, Derek H. Wells, Jeffrey V. Miller, Richard R. Bott, Zachary Q. Beck
Принадлежит: DANISCO US INC, Goodyear Tire and Rubber Co

The present invention provides a three-dimensional structures of P. tremuloides isoprene synthase and P. alba isoprene synthase. The invention also provides methods of using the three-dimensional structure to design isoprene synthases with improved activity for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.

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Номер записи: 5
06-01-2022 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCTION OF MYRCENE

Номер: US20220002735A1
Автор: MAIN Andrew, WOJCIECHOWSKI Grzegorz, Yang Yue, Zhao Lishan
Принадлежит:

Provided herein are compositions and methods for producing myrcene by culturing genetically modified microbial host cells that express a myrcene synthase and optionally a geranyl pyroplosphate synthase. Also provided herein are isolated nucleic acid molecules that encode myrcene synthase variants derived from the species myrcene synthase, which comprise one or more amino acid substitutions that improve in vivo performance of myrcene synthase in genetically modified microbial host cells. Also provided herein are isolated myrcene synthase variants that exhibit an improved activity for converting geranyl diphosphate into myrcene. 137.-. (canceled)38. A genetically modified microbial host cell comprising:{'i': 'Ocimum', 'claim-text': (i) an amino acid sequence of SEQ ID NO: 2; or', '(ii) an amino acid sequence that has at least 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%), at least about 97%, at least about 8%, at least about 99% sequence identity to SEQ ID NO: 2; and, '(a) a heterologous nucleic acid molecule encoding an species myrcene synthase that comprises(b) a heterologous nucleic acid molecule encoding a geranyl pyrophosphate synthase.39. The genetically modified microbial host cell of claim 38 , wherein the heterologous nucleic acid encodes a geranyl pyrophosphate synthase derived from a bacterium.40Streptomyces aculeolatus. The genetically modified host cell of claim 39 , wherein the geranyl pyrophosphate synthase is derived from a geranyl pyrophosphate synthase.41Streptomyces aculeolatus. The genetically modified host cell of claim 40 , wherein the geranyl pyrophosphate synthase comprises an amino acid sequence having SEQ ID NO: 7 or a variant thereof that has at least about 70% claim 40 , at least about 75% claim 40 , at least about 80% claim 40 , at least about 85% claim 40 , at least about 90% claim 40 , at least about 95% claim 40 , at least about 96% claim 40 , at least about ...

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Номер записи: 6
03-01-2019 дата публикации

METHODS, SYNTHETIC HOSTS AND REAGENTS FOR THE BIOSYNTHESIS OF HYDROCARBONS

Номер: US20190002927A1
Автор: FOSTER Alexander Brett, Pattenden Leonard Keith, Rand Jonathan David, ROBERTS Ana Teresa dos Santos Brito Mendes, Shaw Andrew, Taylor Mark Paul
Принадлежит:

Systems, networks, methods, compositions and recombinant hosts for biosynthesizing hydrocarbons from a feedstock, such as gas, are provided. 1: A method for biosynthesising a hydrocarbon in a recombinant host , said method comprising:providing a fermentation reactor, the fermentation reactor comprising at least one recombinant host, wherein said recombinant host comprises an exogenous nucleic acid sequence encoding a polypeptide having an enzyme activity of EC 2.2.1.7, andproviding a stream comprising a gas or a biological or nonbiological feedstock to the fermentation reactor, andoperating the fermentation reactor at conditions for said recombinant host to metabolize the gas or feedstock and produce the hydrocarbon.3: The method according to claim 1 , wherein said polypeptide having an enzyme activity of EC 2.2.1.7 converts glyceraldehyde-3-phosphate and pyruvate to 1 deoxy-d-xylulose-phosphate.4: The method according to claim 1 , wherein said polypeptide having an enzyme activity of EC 2.2.1.7 is:(i) encoded by a nucleic acid sequence having at least 49% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:4 or 5 or a functional fragment thereof;(ii) encoded by a nucleic acid sequence comprising the nucleic acid sequence set forth in SEQ ID NO:4 or 5 or a functional fragment thereof;(iii) has at least 49% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 2 or a functional fragment thereof; or(iv) comprises the amino acid sequence set forth in SEQ ID NO: 1 or 2 or a functional fragment thereof.57-. (canceled)8: The method according to claim 1 , wherein the recombinant host further comprises an exogenous nucleic acid sequence encoding a polypeptide having an enzyme activity of EC 4.2.3.27.10: The method according to claim 8 , wherein said polypeptide having an enzyme activity of EC 4.2.3.27 is:(i) encoded by a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 6 or ...

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Номер записи: 7
01-01-2015 дата публикации

Valencene Synthase Polypeptides, Encoding Nucleic Acid Molecules And Uses Thereof

Номер: US20150007368A1
Автор: Dayal Saran, Grace Eunyoung Park
Принадлежит: Allylix Inc

Provided are valencene synthase polypeptides, nucleic acid molecules encoding the valencene synthases, host cells containing the nucleic acids and methods for producing products whose production is catalyzed by the polypeptides. Also provided are methods for producing valencene and nootkatone.

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Номер записи: 8
20-01-2022 дата публикации

Efficient Production of Steviol Glycosides in Recombinant Hosts

Номер: US20220017932A1
Автор: Anderson Iben Nordmark, Dalgaard Mikkelsen Michael, Douchin Veronique, Hansen Jorgen, Simon Ernesto
Принадлежит:

Recombinant microorganisms are disclosed that produce steviol glycosides and have altered expression of one or more endogenous transporter or transcription factor genes, or that overexpress one or more heterologous transporters, leading to increased excretion of steviol glycosides of interest. 1. A recombinant microorganism capable of producing a steviol glycoside in a cell culture ,wherein the microorganism has a modified expression of at least one endogenous transporter gene and at least one endogenous transcription factor gene that regulates expression of the at least one endogenous transporter gene,wherein the modified expression comprises reduced expression or activity of the at least one endogenous transporter gene, having at least 80% sequence identity to the amino acid sequence set forth in SEQ ID NO:113;wherein the reduced expression is produced in the microorganism by disrupting or deleting the gene locus for the at least one endogenous transporter gene;wherein the microorganism further expresses at least one heterologous transporter gene;wherein the steviol glycoside is Rebaudioside A, Rebaudioside B, Rebaudioside D, Rebaudioside E, Rebaudioside M, or an isomer thereof.2. The recombinant microorganism of claim 1 , wherein the endogenous and/or the heterologous transporter gene encodes an ATP-Binding Cassette (ABC) transporter.3Stevia. The recombinant microorganism of claim 1 , wherein the heterologous transporter gene is a transporter gene.4. The recombinant microorganism of claim 1 , further comprising a gene encoding a polypeptide capable of beta 1 claim 1 ,2 glycosylation of the C2′ of the 13-O-glucose claim 1 , 19-O-glucose claim 1 , or both 13-O-glucose and 19-O-glucose of a steviol glycoside; wherein the gene has a copy number of 2 or more; and wherein the polypeptide comprises a polypeptide having at least 90% sequence identity to one of the amino acid sequences set forth in any one of SEQ ID NOs:51 claim 1 , 54 claim 1 , 55 claim 1 , 86 claim 1 , ...

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Номер записи: 9
12-01-2017 дата публикации

Methods for Isoprene and Pinene Production in Cyanobacteria

Номер: US20170009260A1
Автор: Kallas Toivo, Nelson Matthew, Singsaas Eric
Принадлежит:

Methods of isoprenoid production are provided by the present invention. In particular, transgenic sp. PCC 7002 cyanobacteria and methods for producing isoprene and pinene using a host transgenic sp. PCC 7002 cyanobacterium are provided. 1. A method for pinene production , the method comprising the steps of:{'i': 'Synechococcus', '(a) obtaining a host transgenic sp. PCC 7002 cyanobacterium comprising one or more transgenes encoding geranyl diphosphate synthase (GPPS), and mono-terpene synthase (mono-TPS); and'}{'sup': −1', '−1, '(b) observing pinene production by the cyanobacterium, wherein pinene is produced at a rate of at least about 330 μg gDWh.'}2. The method of claim 1 , wherein pinene is produced at a rate of at least about 660 μg gDWh.3. The method of claim 1 , wherein pinene is produced at a rate of at least about 2000 μg gDWh.4. The method of claim 1 , wherein pinene is produced at a rate of at least about 4000 μg gDWh.5Synechocystis. The method of claim 1 , wherein the cyanobacterium further comprises a promoter derived from sp. PCC 6803.6Synechocystis. The method of claim 5 , wherein the promoter derived from sp. PCC 6803 is PcpcB.7Synechocystis. The method of claim 5 , wherein the promoter derived from sp. PCC 6803 is a synthetic PpsaA/B promoter.8. The method of claim 1 , wherein the one or more transgenes comprises codons preferred for expression in the cyanobacterium.9Artemisia. The method of claim 1 , wherein the one or more transgenes encodes a protein identical to that isolated from an species.10ArtemisiaArtemisia annua.. The method of claim 9 , wherein the protein isolated from an species is mono-TPS from11. The method of claim 1 , wherein the cyanobacterium of step (a) further comprises at least one transgene selected from the group consisting of a transgene encoding hydroxymethylbutenyl diphosphate reductase (HDR) and 1-deoxy-D-xylulose-5-phosphate synthase (DXS).12. The method of claim 1 , wherein pinene is produced under high COconditions.13. ...

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Номер записи: 10
09-01-2020 дата публикации

SANTALENE SYNTHASE

Номер: US20200010822A1
Автор: BEEKWILDER Martinus Julius, Bosch Hendrik Jan, LENTZEN Georg Friedrich, MELILLO Elena, VAN HOUWELINGEN Adèle Margaretha Maria Liduina, Wisselink Hendrik Wouter
Принадлежит:

The invention is directed to a santalene synthase, to a nucleic acid encoding said santalene synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing santalene, to a method of preparing santalol and to a method of preparing a santalene synthase. The invention is further directed to an antibody specific for the santalane synthase. 1. Santalene synthase comprising an amino acid sequence as shown in SEQ ID NO:3 or a functional homologue thereof, said homologue being a santalene synthase comprising an amino acid sequence which has a sequence identity of at least 60% with SEQ ID NO: 3.2. Santalene synthase according to claim 1 , having at least 65% claim 1 , at least 75% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , or at least 98% sequence identity with SEQ ID NO: 3.3. Nucleic acid claim 1 , comprising a nucleic acid sequence encoding a santalene synthase according to claim 1 , or a complementary sequence thereof.4. Nucleic acid according to claim 3 , wherein the nucleic acid comprises a nucleic acid sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 2 claim 3 , or a nucleic acid sequence having a sequence identity of at least 60% claim 3 , at least 65% claim 3 , at least 75% claim 3 , at least 85% claim 3 , at least 90% or at least 95% with a sequence shown in SEQ ID NO: 1 or SEQ ID NO: 2 claim 3 , or a complementary sequence of any of these sequences.5. Expression vector comprising a nucleic acid according to .6. A host cell claim 3 , which may be an organism per se or part of a multi-cellular organism claim 3 , said host cell comprising an expression vector comprising a heterologous nucleic acid sequence according to .7Rhodobacter, ParacoccusEscherichia.. A host cell according to claim 6 , wherein the host cell is a bacterial cell selected from the group of Gram negative bacteria claim 6 , in particular from the group of and8Aspergillus, Blakeslea, ...

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Номер записи: 11
21-01-2016 дата публикации

Pest resistant plants

Номер: US20160017364A1
Автор: Michael Albertus Haring, Petronella Martina Bleeker, Robert Cornelis Schuurink
Принадлежит: Keygene NV

The disclosure provides an isolated nucleic acid molecule encoding a 7-epizingiberene synthase, a chimeric gene comprising said nucleic acid molecule, vectors comprising the same, as well as isolated 7-epizingiberene synthase proteins themselves. In addition, transgenic plants and plant cells comprising a gene encoding a 7-epizingiberene synthase, optionally integrated in its genome, and methods for making such plants and cells, are provided. Especially Solanaceae plants and plant parts (seeds, fruit, leaves, etc.) with enhanced insect pest resistance are provided.

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Номер записи: 12
21-01-2016 дата публикации

COMPOSITIONS AND METHODS FOR BIOLOGICAL PRODUCTION OF ISOPRENE

Номер: US20160017374A1
Автор: Leonard Effendi, Minshull Jeremy, Ness Jon Edward, Purcell Thomas Joseph
Принадлежит:

The present disclosure provides compositions and methods for biologically producing isoprene using methanotrophic bacteria that utilize carbon feedstock, such as methane or natural gas.

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Номер записи: 13
17-01-2019 дата публикации

METHODS, CELLS AND REAGENTS FOR PRODUCTION OF ISOPRENE, DERIVATIVES AND INTERMEDIATES THEREOF

Номер: US20190017076A1
Автор: Conradie Alex Van Eck
Принадлежит:

This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy-3-methylglutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methylglutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene. 112-. (canceled)13. A non-naturally occurring host capable of producing 3-hydroxy-3-methylglutaryl-CoA , said host comprising:(a) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.- enzyme; or at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme;', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.2.1.2. enzyme; or (c) both (a) and (b); and, '(b) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.74 or EC 4.1.1.43 enzyme;'}at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.3.8.4 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.4.1.4 enzyme; andat least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.2.1.18 enzyme.14. The host of claim 13 , wherein said host is capable of producing isoprene and comprises:at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.1.1.34 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.1.36 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.4.2 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.33 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 5.3.3.2 ...

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Номер записи: 14
16-01-2020 дата публикации

Terpene Synthases for Biofuel Production and Methods Thereof

Номер: US20200017885A1
Автор: Davis Ryan Wesley, Gladden John Michael, Wu Benjamin Chiau-pin, Wu Weihua
Принадлежит:

The present invention relates to terpene synthases capable of degrading precursors into biofuel compounds, such as terpenoid compounds. In one instance, a transformed organism can include such terpene synthases, as well as vectors encoding such synthases. Methods of employing such synthases and organisms are also described herein. 1. A method of treating a biomass , the method comprising: an exogenous terpenoid precursor, an exogenous enzyme configured to synthesize a terpenoid precursor, or a nucleic acid encoding the exogenous enzyme; and', 'an exogenous terpene synthase or a nucleic acid encoding the exogenous terpene synthase, wherein the exogenous terpene synthase is selected from the group consisting of a pinene synthase, a guaiene synthase, a pinene and guaiene synthase, a caryophyllene synthase, a chamigrene synthase, a chamigrene and pinene synthase, a gurjunene synthase, a gurjunene and pinene synthase, a gumunene synthase, a selinene synthase, and an isoledene synthase, or a bifunctional synthase of any of these; and, 'exposing the biomass to one or more organisms of an isolated, genetically engineered organism, wherein the organism comprisesisolating one or more terpenoid compounds.2. The method of claim 1 , wherein the exogenous terpenoid precursor is selected from the group consisting of mevalonate claim 1 , dimethylallyl pyrophosphate claim 1 , isopentenyl pyrophosphate claim 1 , farnesyl pyrophosphate claim 1 , geranyl pyrophosphate claim 1 , and geranylgeranyl pyrophosphate claim 1 , or a salt thereof.3. The method of claim 1 , wherein the organism is configured to produce one or more terpenoid compounds selected from the group consisting of a monoterpene claim 1 , a sesquiterpene claim 1 , and a diterpene.4. The method of claim 1 , wherein the nucleic acid encoding the exogenous enzyme and/or the nucleic acid encoding the exogenous terpene synthase is provided as a plasmid vector.5. The method of claim 1 , wherein the exogenous enzyme is selected ...

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Номер записи: 15
25-01-2018 дата публикации

METHODS OF PRODUCING FOUR CARBON MOLECULES

Номер: US20180023097A1
Автор: Botes Adriana Leonora, Chen Changlin, Pearlman Paul S.
Принадлежит: INVISTA North America S.a.r.l.

Disclosed are methods for producing butadiene from one or more of several diverse feedstocks including bioderived feedstocks, renewable feedstocks, petrochemical feedstocks and natural gas. 1. A method for biosynthesizing butadiene comprising:fermenting in an organism in a fermentation broth, a C4-precursor, wherein said C4-precursor comprises butanol, butanediol, or both, or derivatives thereof; (a) converting butanediol to butenol, or', '(b) converting butanol to butenol;, 'converting C4-precursor in the fermentation broth into butenol byconverting the butenol into 1,3 butadiene; andoptionally isolating said 1,3-butadiene.2. The method of claim 1 , wherein the fermentation broth comprises a biologically derived fermentable feedstock or a nonbiologically derived fermentable feedstock.3. The method of claim 2 , wherein the biologically derived feedstock comprises glycerol claim 2 , synthesis gas claim 2 , synthesis gas from biomass claim 2 , sugar claim 2 , sugar from a food stuff claim 2 , sugar from a non-food stuff claim 2 , or combinations thereof.4. The method of claim 3 , wherein the sugar is sucrose claim 3 , glucose claim 3 , or combinations thereof.5. The method of claim 3 , wherein the sugar from a non-food stuff is cellulosic or hemi-cellulosic sugar or combinations thereof.6. (canceled)7. The method of claim 2 , wherein the non-biologically derived feedstock is synthesis gas from coal claim 2 , natural gas claim 2 , combustion off-gases claim 2 , municipal waste claim 2 , petrochemical claim 2 , or combinations thereof.8. (canceled)9. The method of claim 1 , wherein the organism is a prokaryote or eukaryote.10. The method of claim 1 , wherein the C4 precursor is a hydroxylated C4 molecule.11. The method of claim 1 , wherein converting the butanediol to butenol requires at least one polypeptide having an activity of at least one enzyme.12. The method of claim 11 , wherein the at least one enzyme is a dehydratase claim 11 , desaturase or hydrolase enzyme. ...

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Номер записи: 16
25-01-2018 дата публикации

NUCLEIC ACID, FUSION PROTEIN, RECOMBINED CELL, AND ISOPRENE OR CYCLIC TERPENE PRODUCTION METHOD

Номер: US20180023098A1
Автор: FURUTANI Masahiro, KAWABATA Kazufumi, NISHIYAMA Norihide
Принадлежит: Sekisui Chemical Co., Ltd.

Provided is a nucleic acid encoding a fusion protein, the fusion protein including a first protein selected from the group consisting of isoprene synthase and cyclic terpene synthase, and a FKBP family protein linked to the first protein. Provided is a fusion protein encoded by the nucleic acid. Provided is a recombinant cell including the nucleic acid and expressing the fusion protein. Further provided is a recombinant cell including a first nucleic acid encoding the first protein and a second nucleic acid encoding the FKBP family protein, and expressing the first protein and the FKBP family protein. As a host cell, a syngas-assimilating bacterium or a methanol assimilating bacterium can be used. 1. A nucleic acid encoding a fusion protein , the fusion protein comprising:a first protein selected from the group consisting of isoprene synthase and cyclic terpene synthase; anda FKBP family protein linked to the first protein.2. The nucleic acid according to claim 1 , wherein the first protein is isoprene synthase.3. The nucleic acid according to claim 2 , wherein the isoprene synthase is any one of the following (a-1) to (a-3):(a-1) a protein consisting of the amino acid sequence of SEQ ID NO: 2,(a-2) a protein consisting of the amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence of SEQ ID NO: 2, and having isoprene synthase activity, and(a-3) a protein consisting of an amino acid sequence having homology of 90% or more with the amino acid sequence of SEQ ID NO: 2, and having isoprene synthase activity.4. The nucleic acid according to claim 1 , wherein the first protein is cyclic monoterpene synthase.5. The nucleic acid according to claim 4 , wherein the cyclic monoterpene synthase is phellandrene synthase.6. The nucleic acid according to claim 4 , wherein the cyclic monoterpene synthase is any one of the following (b-1) to (b-3):(b-1) a protein consisting of the amino acid sequence of SEQ ID NO: 4,(b-2) a ...

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Номер записи: 17
01-02-2018 дата публикации

GENETICALLY MODIFIED HOST CELLS AND USE OF SAME FOR PRODUCING ISOPRENOID COMPOUNDS

Номер: US20180030481A1
Автор: Keasling Jay D., Kirby James, Paradise Eric M.
Принадлежит:

The present invention provides genetically modified eukaryotic host cells that produce isoprenoid precursors or isoprenoid compounds. A subject genetically modified host cell comprises increased activity levels of one or more of mevalonate pathway enzymes, increased levels of prenyltransferase activity, and decreased levels of squalene synthase activity. Methods are provided for the production of an isoprenoid compound or an isoprenoid precursor in a subject genetically modified eukaryotic host cell. The methods generally involve culturing a subject genetically modified host cell under conditions that promote production of high levels of an isoprenoid or isoprenoid precursor compound. 1. A genetically modified eukaryotic host cell that produces an isoprenoid or an isoprenoid precursor compound via a mevalonate pathway , the genetically modified eukaryotic host cell comprising genetic modifications that provide for:a) an increased level of activity of one or more mevalonate pathway enzymes,b) an increased level of prenyltransferase activity, andc) a decreased level of squalene synthase activitywherein the genetic modifications provide for production of an isoprenoid or an isoprenoid precursor compound at a level that is at least about 50% higher than the level of the isoprenoid or isoprenoid precursor compound in a control cell not comprising the genetic modifications.2. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is farnesyl pyrophosphate synthase.3. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranyl pyrophosphate synthase.4. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranylgeranyl pyrophosphate synthase.5. The genetically modified eukaryotic host cell of claim 1 , wherein the genetically modified eukaryotic host cell is a yeast cell.6Saccharomyces cerevisiae.. The genetically modified host cell of claim 5 , wherein the ...

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Номер записи: 18
31-01-2019 дата публикации

SYSTEMIC SYNTHESIS AND REGULATION OF L-DOPA

Номер: US20190032079A1
Автор: McDonald Michael
Принадлежит:

The present invention relates to an expression system for enzyme replacement therapy with the aim of obtaining or maintaining a steady level of L-DOPA in the blood of an individual, achieved through systemic administration of the expression system. The invention is thus useful in the treatment of catecholamine deficient disorders, such as dopamine deficient disorders including Parkinson's Disease. 1. (canceled)2. (canceled)3. (canceled)4. (canceled)5. (canceled)6. (canceled)7. (canceled)8. (canceled)9. (canceled)10. (canceled)11. (canceled)12. (canceled)13. (canceled)14. (canceled)15. (canceled)16. (canceled)17. (canceled)18. (canceled)19. (canceled)20. (canceled)21. (canceled)22. (canceled)23. (canceled)24. (canceled)25. (canceled)26. (canceled)27. (canceled)28. (canceled)29. (canceled)30. (canceled)31. (canceled)32. (canceled)33. (canceled)34. (canceled)35. (canceled)36. (canceled)37. (canceled)38. (canceled)39. (canceled)40. (canceled)41. (canceled)42. (canceled)43. (canceled)44. (canceled)45. (canceled)46. (canceled)47. (canceled)48. (canceled)49. (canceled)50. (canceled)51. (canceled)52. (canceled)53. (canceled)54. (canceled)55. (canceled)56. (canceled)57. (canceled)58. (canceled)59. (canceled)60. (canceled)61. (canceled)62. (canceled)63. (canceled)64. (canceled)65. (canceled)66. (canceled)67. (canceled)68. (canceled)69. (canceled)70. (canceled)71. (canceled)72. (canceled)73. (canceled)74. (canceled)75. (canceled)76. (canceled)77. (canceled)78. (canceled)79. (canceled)80. (canceled)81. (canceled)82. (canceled)83. (canceled)84. (canceled)85. (canceled)86. (canceled)87. (canceled)88. (canceled)89. (canceled)90. (canceled)91. (canceled)92. (canceled)93. (canceled)94. (canceled)95. (canceled)96. (canceled)97. (canceled)98. (canceled)99. (canceled)100. (canceled)101. (canceled)102. (canceled)103. (canceled)104. (canceled)105. (canceled)106. (canceled)107. (canceled)108. (canceled)109. (canceled)110. (canceled)111. (canceled)112. (canceled)113. (canceled)114. ( ...

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Номер записи: 19
05-02-2015 дата публикации

MICROORGANISMS FOR THE PRODUCTION OF 5-HYDROXYTRYPTOPHAN

Номер: US20150037849A1
Автор: Forster Jochen, Knight Eric Michael, Luo Hao, Zhu Jiangfeng
Принадлежит:

Recombinant microbial cells and methods for producing 5-hydroxytryptophan (5HTP) using such cells are described. More specifically, the recombinant microbial cell comprises an exogenous gene encoding an L-tryptophan hydroxylase, and means for providing tetrahydrobiopterin (THB). Related sequences and vectors for use in preparing such recombinant microbial cells are also described. 1. A recombinant microbial cell comprising an exogenous nucleic acid sequence encoding an L-tryptophan hydroxylase (TPH) (EC 1.14.16.4) , and exogenous nucleic acid sequences encoding enzymes of at least one pathway for producing tetrahydrobiopterin (THB).2. The recombinant microbial cell of claim 1 , comprising exogenous nucleic acid sequences encoding enzymes of a first pathway producing THB from guanosin triphosphate (GTP) claim 1 , of a second pathway regenerating THB from 4a-hydroxytetrahydrobiopterin claim 1 , or of both the first and the second pathway.3. The recombinant microbial cell of any one of the preceding claims claim 1 , comprising exogenous nucleic acid sequences encoding(a) optionally, a GTP cyclohydrolase I (EC 3.5.4.16);(b) a 6-pyruvoyl-tetrahydropterin synthase (EC 4.2.3.12); and(c) a sepiapterin reductase (EC 1.1.1.153).4. The recombinant microbial cell of any one of the preceding claims claim 1 , comprising exogenous nucleic acid sequences encoding(a) a 4a-hydroxytetrahydrobiopterin dehydratase (EC 4.2.1.96); and(b) optionally, a dihydropteridine reductase (EC 1.5.1.34).5. The recombinant microbial cell of any one of the preceding claims claim 1 , wherein each one of said exogenous nucleic acid sequences is operably linked to an inducible claim 1 , a regulated or a constitutive promoter.6. The recombinant microbial cell of any one of the preceding claims claim 1 , which comprises a mutation providing for reduced tryptophanase activity.7. The recombinant microbial cell of any one of the preceding claims claim 1 , which is derived from a microbial host cell which is a ...

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Номер записи: 20
26-02-2015 дата публикации

ENGINEERING SALT TOLERANCE IN PHOTOSYNTHETIC MICROORGANISMS

Номер: US20150056707A1
Автор: Fang Su-Chiung, Mendez Michael, RICHARD Stephane
Принадлежит: SAPPHIRE ENERGY, INC.

Provided herein are compositions and methods for engineering salt tolerance and producing products by photosynthetic organisms. The photosynthetic organisms can be genetically modified to be salt tolerant as compared to an unmodified organism and to produce useful products. The methods and compositions of the disclosure are useful in many therapeutic and industrial applications. 1. A non-vascular photosynthetic organism transformed with an exogenous polynucleotide encoding a protein of SEQ ID NO. 36 or at least 90% sequence identity to SEQ ID NO. 36 , wherein said transformed non-vascular photosynthetic organism has a greater salt tolerance than said organism not transformed with said exogenous polynucleotide.2. The organism of claim 1 , wherein said organism is an alga.3Nannochloropsis, Chlamydomonas, ScenedesmusDunaliella.. The organism of claim 2 , wherein said organism is an alga of the genus claim 2 , or4. The organism of claim 1 , wherein said organism is a cyanobacterium.5Spirulina.. The organism of claim 4 , wherein said cyanobacterium is of the genus6. The organism of claim 1 , wherein said organism is transformed with a second exogenous polynucleotide.7. The organism of claim 6 , wherein said second exogenous polynucleotide encodes for a chaperonin claim 6 , an antioxidant claim 6 , a biodegradative enzyme claim 6 , exo-β-glucanase claim 6 , endo-β-glucanase claim 6 , β-glucosidase claim 6 , endoxylanase claim 6 , lignase claim 6 , a flocculating moiety claim 6 , a botryococcene synthase claim 6 , a limonene synthase claim 6 , a 1 claim 6 ,8 cineole synthase claim 6 , a α-pinene synthase claim 6 , a camphene synthase claim 6 , a (+)-sabinene synthase claim 6 , a myrcene synthase claim 6 , an abietadiene synthase claim 6 , a taxadiene synthase claim 6 , a farnesyl pyrophosphate synthase claim 6 , an amorphadiene synthase claim 6 , a (E)-α-bisabolene synthase claim 6 , a diapophytoene synthase claim 6 , a diapophytoene desaturase claim 6 , a transporter ...

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Номер записи: 21
10-03-2022 дата публикации

Recombinant Production of Steviol Glycosides

Номер: US20220073960A1
Автор: Blom, Hansen Esben Halkjaer, Hansen Jorgen, HICKS Paula M., Hougton-Larsen Jens, Kishore Ganesh M., Mikkelsen Michael Dalgaard, Motion Michael, Sr. Charlotte, Tavares Sabina
Принадлежит:

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements. 1. A method for producing Rebaudioside D (RebD) , Rebaudioside E (RebE) , or a mixture thereof , comprising contacting a precursor steviol glycoside having a 13-O-glucose , a 19-O-glucose , or both the 13-O-glucose and the 19-O-glucose with a polypeptide capable of beta 1 ,2 glycosylation of the C-2′ of the 13-O-glucose , 19-O-glucose , or both the 13-O-glucose and the 19-O-glucose of the precursor steviol glycoside and a UDP-glucose in a reaction mixture under suitable conditions for the transfer of one or more glucose moiety to the C2′ of the 13-O-glucose , 19-O-glucose or both 13-O-glucose and 19-O-glucose in the precursor steviol glycoside; thereby producing RebD , RebE , or a mixture thereof;wherein the polypeptide comprises an amino acid motif AA1-AA2-AA3-AA4-AA5-AA6-AA7, corresponding to residues 20-26 in SEQ ID NO:5; and{'claim-text': [{'sub': '1', '#text': 'AAis Proline'}, {'sub': '2', '#text': 'AAis one aromatic amino acid;'}, 'AA3 is one large hydrophobic amino acid;', 'AA4 is one small amino acid;', 'AA5 is one amino acid;', 'AA6 is one small amino acid; and', 'AA7 is Histidine.'], '#text': 'wherein:'}2. The method of claim 1 , wherein AA2 is:{'sup': '3', 'sub': 'R', '#text': '(a) one amino acid having a van der Waals volume ≥130 Åand a side chain hydrophobicity ≥60 Δt;'}{'sup': '3', 'sub': 'R', '#text': '(b) one amino acid having a van der Waals volume ≥130 Åand a side chain hydrophobicity ≥80 Δt;'}(c) Tryptophan, Phenylalanine, or Tyrosine;(d) Tryptophan or Phenylalanine; or(e) Tryptophan.3. The method of claim 1 , wherein AA3 is:{'sup': '3', 'sub': 'R ...

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Номер записи: 22
04-03-2021 дата публикации

MICROBIAL FERMENTATION FOR THE PRODUCTION OF TERPENES

Номер: US20210062229A1
Автор: Koepke Michael
Принадлежит:

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways. 1ClostridiumMoorella.. A recombinant C1-fixing bacteria capable of producing mevalonic acid , or a terpene precursor , from a carbon source comprising a nucleic acid encoding a group of exogenous enzymes comprising thiolase , 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase , and HMG-CoA reductase , wherein the bacteria are or2. The bacteria according to claim 1 , further comprising a nucleic acid encoding a group of enzymes comprising mevalonate kinase claim 1 , phosphomevalonate kinase claim 1 , and mevalonate diphosphate decarboxylase.3. The bacteria according to claim 2 , wherein the terpene precursor is isopentenyl diphosphate.4. The bacteria according to claim 2 , further comprising a nucleic acid encoding an exogenous enzyme selected from the group consisting of isopentenyl diphosphate isomerase and geranyltranstransferase.5. The bacteria according to claim 2 , further comprising a nucleic acid encoding both exogenous enzymes isopentenyl diphosphate isomerase and geranyltranstransferase.6. The bacteria according to claim 4 , wherein the terpene precursor is dimethylallyl pyrophosphate or geranyl pyrophosphate.7. The bacteria according to claim 4 , further comprising a nucleic acid encoding an exogenous enzyme comprising isoprene synthase.8. The bacteria according to claim 4 , wherein the terpene precursor is farnesyl pyrophosphate.9. The bacteria according to claim 4 , further comprising a nucleic acid ...

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Номер записи: 23
08-03-2018 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCING ISOPRENE FREE OF C5 HYDROCARBONS UNDER DECOUPLING CONDITIONS AND/OR SAFE OPERATING RANGES

Номер: US20180066287A1
Автор: CALABRIA Anthony R., Cervin Marguerite A., Chotani Gopal K., La Duca Richard, McAuliffe Joseph C., Miller Michael C., SABO Timothy A., SANFORD Karl J., Spring Erin L., Whited Gregory M.
Принадлежит:

The invention features methods for producing isoprene from cultured cells wherein the cells in the stationary phase. The invention also provides compositions that include these cultured cells and/or increased amount of isoprene. The invention also provides for systems that include a non-flammable concentration of isoprene in the gas phase. Additionally, the invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity. 117-. (canceled)18. A biologically-produced composition comprising:(a) greater than about 2 mg of isoprene; and(b) two or more compounds selected from the group consisting of 2-heptanone, 6-methyl-5-hepten-2-one, 2,4,5-trimethylpyridine, 2,3,5-trimethylpyrazine, citronellal, acetaldehyde, methanethiol, methyl acetate, 1-propanol, diacetyl, 2-butanone, 2-methyl-3-buten-2-ol, ethyl acetate, 2-methyl-1-propanol, 3-methyl-1-butanal, 3-methyl-2-butanone, 1-butanol, 2-pentanone, 3-methyl-1-butanol, ethyl isobutyrate, 3-methyl-2-butenal, butyl acetate, 3-methylbutyl acetate, 3-methyl-3-buten-1-yl acetate, (E)-3,7-dimethyl-1,3,6-octatriene, (Z)-3,7-dimethyl-1,3,6-octatriene, and 2,3-cycloheptenolpyridine; wherein the amount of at least one of the two or more compounds relative to the amount of the isoprene is between about 0.01% to 105% (w/w).1931-. (canceled)32. The composition of claim 18 , wherein at least a portion of the isoprene is in the gas phase.33. The composition of claim 18 , wherein the amount of one of the two or more compounds relative to the amount of the isoprene is between about 0.01% to about 90% (w/w) claim 18 , between about 0.01% to about 80% (w/w) claim 18 , between about 0.01% to about 50% (w/w) claim 18 , between about 0.01% to about 20% (w/w) claim 18 , between about 0.01% to about 10% (w/w) claim 18 , between about 0.02% to about 50% (w/w) claim 18 , between about 0.05% to about 50% (w/w) claim 18 , between about 0.1% to about 50% (w/w) claim 18 , or between about 0.1% to about ...

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Номер записи: 24
15-03-2018 дата публикации

EXTRACELLULAR DITERPENE PRODUCTION

Номер: US20180073050A1
Автор: BOER Viktor Marius, BROERS Nicolette Jasmijn, LAWRENCE Adam G.
Принадлежит:

The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism in a suitable fermentation medium, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol, whereby a diterpene or glycosylated diterpene is produced extracellularly in the fermentation medium; and b. recovering the diterpene or glycosylated diterpene from the fermentation medium. 1. A method for the production of a diterpene or a glycosylated diterpene , which method comprises:a. fermenting a recombinant microorganism in a suitable fermentation medium,wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol,whereby a diterpene or glycosylated diterpene is produced extracellularly in the fermentation medium; andb. recovering the diterpene or glycosylated diterpene from the fermentation medium.2. The method according to claim 1 , wherein the recombinant microorganism comprises one or more nucleotide sequences encoding a polypeptide having UDP-glucosyltransferase activity claim 1 ,whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least one of steviolmonoside, steviolbioside, stevioside or ...

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Номер записи: 25
24-03-2022 дата публикации

GERANYLGERANYL PYROPHOSPHATE SYNTHASES

Номер: US20220090032A1
Автор: BOER Viktor Marius, VAN LEEUWEN Johannes Gustaaf Ernst, ZWARTJENS Priscilla
Принадлежит:

The present invention relates a variant polypeptide having geranylgeranyl pyrophosphate synthase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a geranylgeranyl pyrophosphate synthase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids at positions 92, 100 or 235 said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having geranylgeranyl pyrophosphate synthase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside. 1. A process for preparing steviol or a steviol glycoside , said process comprising fermenting a recombinant host in a suitable fermentation medium , wherein the recombinant host comprises a polynucleotide encoding a variant polypeptide having geranylgeranyl pyrophosphate synthase activity , which variant polypeptide comprises an amino acid sequence which , when aligned with a geranylgeranyl pyrophosphate synthase comprising the sequence of SEQ ID NO:1 , comprises a substitution of an amino acid residue corresponding to any of amino acids at positions92, 100, or 235,said positions being relative to SEQ ID NO:1.2. The process of claim 1 , wherein the variant polypeptide has modified geranylgeranyl pyrophosphate synthase activity as compared with a reference polypeptide having geranylgeranyl pyrophosphate synthase activity claim 1 , and wherein the reference polypeptide comprises the geranylgeranyl pyrophosphate synthase of SEQ ID NO:1 or SEQ ID NO:17.3. The process of claim 1 , wherein the variant polypeptide comprises a substitution relative to SEQ ID NO:1 selected from the group consisting of G92E claim 1 , G92D claim 1 , G92N claim 1 , and G92Q.4. The process of claim 1 , wherein the variant polypeptide comprises a substitution ...

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Номер записи: 26
24-03-2022 дата публикации

PTERIN-DEPENDENT BIOCATALYSTS AND USES THEREOF

Номер: US20220090154A1
Автор: EHRENWORTH Amy M., Peralta-Yahya Pamela
Принадлежит:

Provided herein are biocatalysts and systems thereof for pterin-dependent enzymes and pathways and methods of making and using the same. 1. A biocatalyst comprising:a tetrahydrobiopterin source; anda pterin-dependent enzymatic pathway, where the tetrahydrobioprterin source is biologically coupled to the pterin-dependent enzymatic pathway.2. The biocatalyst of claim 1 , wherein the tetrahydrobiopterin source is a tetrahydrobiopterin synthesis pathway comprising:a GTP cyclohydrase;a pyruvoyl tetrahydropterin synthase; anda sepiapterin reductase.3. The biocatalyst of claim 1 , wherein the pterin-dependent enzymatic pathway comprises at least one enzyme selected from the group consisting of:an amino acid mono-oxygenase, a modified amino acid mono-oxygenase, phenylalanine hydroxylase, tryptophan hydroxylase, nitric oxide synthase, and tyrosine hydroxylase.4. The biocatalyst of claim 1 , wherein said pterin-dependent enzymatic further comprises aa decarboxylase.5. The biocatalyst of claim 4 , wherein the decarboxylase is aromatic-I-amino acid decarboxylase.6. The biocatalyst of claim 1 , further comprising a synthase.7. The biocatalyst of claim 6 , wherein the synthase is a terpene alkaloid synthase claim 6 , strictosidine synthase claim 6 , or a deacetylisoipecoside synthase.8. The biocatalyst of claim 1 , further comprising a tetrahydrobiopterin recycling pathway claim 1 , the tetrahydrobiopterin pathway comprising:a pterin-4a-carbinolamine dehydratase; anda dihydropterin reductase, where the dihydropterin reductase is biologically coupled to the pterin-4a-carbinolamine dehydratase;where the tetrahydrobiopterin pathway is biologically coupled to the pterin-dependent enzymatic pathway.9. An engineered cell comprising:{'claim-text': ['a tetrahydrobiopterin source; and', 'a pterin-dependent enzymatic pathway biologically coupled to the tetrahydrobiopterin source.'], '#text': 'a biocatalyst, the biocatalyst comprising:'}10. The engineered cell of claim 9 , wherein the ...

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Номер записи: 27
19-03-2015 дата публикации

VALENCENE SYNTHASE

Номер: US20150079649A1
Автор: DE JONG Rene M., SONKE Theodorus
Принадлежит: Isobionics B.V.

The present invention relates to a novel valencene synthase, to a nucleic acid encoding such valencene synthase, to a host cell comprising said encoding nucleic acid sequence and to a method for preparing valencene, comprising converting farnesyl diphosphate to valencene in the presence of a valencene synthase according to the invention. 1. Valencene synthase , which valence synthase has an increased productivity towards the conversion of farnesyl diphosphate into valencene (expressed as molar amount of valencene formed per hour) compared to a valencene synthase represented by SEQ ID NO: 2 , or which valencene synthase comprises an amino acid sequence represented by SEQ ID NO: 3 provided that at least one position marked ‘X’ in SEQ ID NO: 3 is different from the corresponding position in SEQ ID NO: 2.2. Valencene synthase according to claim 1 , wherein the valencene synthase has an increased specific productivity claim 1 , increased stability claim 1 , increased product specificity (relative to the conversion of farnesyl diphosphate into Germacrene A) or an increased expression in a host cell claim 1 , compared to a valencene synthase represented by SEQ ID NO: 2.3. Valencene synthase according to claim 2 , wherein the product specificity claim 2 , expressed as the molar ratio valencene formed from farnesyl diphospate to Germacrene A formed from farnesyl diphosphate (under test conditions) claim 2 , is 10 or more claim 2 , preferably 13-30 claim 2 , in particular 15-25.4. Valencene synthase according to wherein the specific productivity of the valencene synthase claim 1 , expressed as the molar amount of valencene formed per hour per amount of valencene synthase is at least 1.5 times claim 1 , preferably 2.0 to 10 times claim 1 , in particular 2.5 to 5 times the specific productivity of the valencene synthase represented by SEQ ID NO: 2.5. Valencence synthase according to wherein the valencene synthase has at least one modification claim 1 , in particular at least ...

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Номер записи: 28
14-03-2019 дата публикации

KAURENOIC ACID HYDROXYLASES

Номер: US20190078105A1
Автор: BOER Viktor Marius, VAN LEEUWEN Johannes Gustaaf Ernst, ZWARTJENS Priscilla
Принадлежит:

The present invention relates a variant polypeptide having kaurenoic acid 13-hydroxylase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids 72, 85, 108, 127, 129, 141, 172, 195, 196, 197, 199, 226, 236, 291, 302, 361 or 464, said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside. 1. A variant polypeptide having kaurenoic acid 13-hydroxylase activity , which variant polypeptide comprises an amino acid sequence which , when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1 , comprises at least one substitution of an amino acid residue corresponding to any of amino acids72, 85, 108, 127, 129, 141, 172, 195, 196, 197, 199, 226, 236, 291, 302, 361 or 464said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity.2. A variant polypeptide according to claim 1 , wherein the modified property is modified kaurenoic acid 13-hydroxylase activity.3. A variant polypeptide according to claim 1 , wherein the reference polypeptide comprises the kaurenoic acid 13-hydroxylase of SEQ ID NO: 1.4. A variant polypeptide according to claim 1 , wherein the variant polypeptide is a non-naturally occurring polypeptide.5. A variant polypeptide according to which comprises additional substitutions.6. A variant polypeptide according to having at least 70% claim 1 , at least 75% claim 1 , at least 80% claim 1 , at least ...

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Номер записи: 29
14-03-2019 дата публикации

Production of Steviol Glycosides in Microorganisms

Номер: US20190078128A9
Автор: Blom Charlotte, Hansen Esben Halkjaer, Hansen Jorgen, HICKS Paula M., Houghton-Larsen Jens, Kishore Ganesh M., Mikkelsen Michael Dalgaard, Motion Michael, Tavares Sabina
Принадлежит:

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements. 1. A recombinant host cell capable of producing steviol , a target steviol glycoside or a target steviol glycoside composition , comprising: 'wherein the polypeptide is capable of transferring a sugar moiety to the C2′ of a glucose in the precursor steviol glycoside;', '(a) a gene encoding a polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of a precursor steviol glycoside;'}and one or more of:(b) a gene encoding a polypeptide capable of glycosylating steviol or the precursor steviol glycoside at its C-13 hydroxyl group; and/or(c) a gene encoding a polypeptide capable of beta 1,3 glycosylation of the C3′ of the 13-O-glucose of the precursor steviol glycoside; and/or(d) a gene encoding a polypeptide capable of glycosylating steviol or the precursor steviol glycoside at its C-19 carboxyl group;wherein at least one of the genes is a recombinant gene.2. The recombinant host cell of claim 1 , wherein:(a) the precursor steviol glycoside is rubusoside, wherein the sugar moiety is glucose, and stevioside is produced upon transfer of the glucose moiety;(b) the precursor steviol glycoside is stevioside, the sugar moiety is glucose, and rebaudioside E is produced upon transfer of the glucose moiety;(c) the precursor steviol glycoside is stevioside, the sugar moiety is glucose, the stevioside is contacted with the polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of the precursor steviol glycoside and a polypeptide ...

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Номер записи: 30
26-03-2015 дата публикации

Genetically Modified Host Cells and Use of Same for Producing Isoprenoid Compounds

Номер: US20150087042A1
Автор: Keasling Jay D., Kirby James, Paradise Eric M.
Принадлежит:

The present invention provides genetically modified eukaryotic host cells that produce isoprenoid precursors or isoprenoid compounds. A subject genetically modified host cell comprises increased activity levels of one or more of mevalonate pathway enzymes, increased levels of prenyltransferase activity, and decreased levels of squalene synthase activity. Methods are provided for the production of an isoprenoid compound or an isoprenoid precursor in a subject genetically modified eukaryotic host cell. The methods generally involve culturing a subject genetically modified host cell under conditions that promote production of high levels of an isoprenoid or isoprenoid precursor compound. 1. A genetically modified eukaryotic host cell that produces an isoprenoid or an isoprenoid precursor compound via a mevalonate pathway , the genetically modified eukaryotic host cell comprising genetic modifications that provide for:a) an increased level of activity of one or more mevalonate pathway enzymes,b) an increased level of prenyltransferase activity, andc) a decreased level of squalene synthase activitywherein the genetic modifications provide for production of an isoprenoid or an isoprenoid precursor compound at a level that is at least about 50% higher than the level of the isoprenoid or isoprenoid precursor compound in a control cell not comprising the genetic modifications.2. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is farnesyl pyrophosphate synthase.3. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranyl pyrophosphate synthase.4. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranylgeranyl pyrophosphate synthase.5. The genetically modified eukaryotic host cell of claim 1 , wherein the genetically modified eukaryotic host cell is a yeast cell.6Saccharomyces cerevisiae.. The genetically modified host cell of claim 5 , wherein the ...

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Номер записи: 31
29-03-2018 дата публикации

Enzyme for biosynthesis of isoprene and isopentenyl, and mutant thereof

Номер: US20180087040A1
Автор: Deyong GE, Yanfen XUE, Yanhe Ma
Принадлежит: Institute of Microbiology of CAS

The invention provides a polypeptide capable of using 4 -hydroxy- 3 -methyl-but- 2 -enyl pyrophosphate (HMBPP) as a substrate to produce isoprene, a nucleic acid encoding the polypeptide, and a vector and a cell comprising the nucleic acid. In addition, the invention also provides a method for producing isoprene using the polypeptide, and a method for preparing the polypeptide.

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Номер записи: 32
09-04-2015 дата публикации

METHOD FOR PRODUCING PATCHOULOL AND 7-EPI-ALPHA-SELINENE

Номер: US20150099283A1
Автор: Deguerry Fabienne, Schalk Michel
Принадлежит: FIRMENICH SA

A method of producing patchoulol and 7-epi-α-selinene by contacting at least one polypeptide with farnesyl phyrophosphate (fpp). The method may be carried out in vitro or in vivo to produce patchoulol and 7-epi-α-selinene, compounds which can be useful in the field of perfumery. 1. A method for producing patchoulol and 7-epi-α-selinene comprisinga) contacting FPP with at least one polypeptide having a patchoulol and 7-epi-α-selinene synthase activity and comprising an amino acid sequence at least 50% identical to SEQ ID NO:1;b) optionally, isolating the patchoulol and 7-epi-α-selinene produced in step a).2. The method of claim 1 , wherein step a) comprises cultivating a non-human host organism or cell capable of producing FPP and transformed to express at least one polypeptide comprising an amino acid sequence at least 50% identical to SEQ ID NO:1 and having a patchoulol and 7-epi-α-selinene synthase activity claim 1 , under conditions conducive to the production of patchoulol and 7-epi-α-selinene.3. The method of claim 2 , which further comprises claim 2 , prior to step a) claim 2 , transforming a non human host organism or cell capable of producing FPP with at least one nucleic acid encoding a polypeptide comprising an amino acid sequence at least 50% identical to SEQ ID NO:1 and having a patchoulol and 7-epi-α-selinene synthase activity claim 2 , so that said organism expresses said polypeptide.4. The method of claim 3 , wherein the nucleic acid comprises a nucleotide sequence at least 50% claim 3 , preferably at least 70% claim 3 , preferably at least 90% identical to SEQ ID NO:2 or the complement thereof claim 3 , or the nucleic acid comprises or consists of the nucleotide sequence SEQ ID NO:2 or the complement thereof.5. The method of claim 2 , wherein the non-human host organism is a plant claim 2 , a prokaryote or a fungus claim 2 , a microorganism such as a bacteria or yeast claim 2 , or a plant cell or a fungal cell.6E. coliSaccharomyces cerevisiae.. The ...

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Номер записи: 33
05-04-2018 дата публикации

Production of fragrant compounds

Номер: US20180094281A1
Автор: Fabienne Deguerry, Michel Schalk, Pan Li, Qi Wang, Xiufeng He
Принадлежит: FIRMENICH SA

Provided herein is an isolated polypeptide from Juniperus virginiana, Platycladus orientalis ‘Beverleyensis’ or Platycladus orientalis comprising a (+)-cedrol or a (−)-thujopsene synthase. Further provided herein is an isolated nucleic acid molecule from Juniperus virginiana, Platycladus orientalis ‘Beverleyensis’ or Platycladus orientalis encoding a (+)-cedrol or (−)-thujopsene synthase. Further provided herein are methods of producing (+)-cedrol or (−)-thujopsene.

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Номер записи: 34
05-04-2018 дата публикации

PRODUCTION OF TERPENES AND TERPENOIDS

Номер: US20180094286A1
Автор: Ajikumar Parayil Kumaran, GLIEDER Anton, Vogl Thomas
Принадлежит: Technische Universitaet Graz

The present invention relates to a nucleic acid construct comprising a nucleic acid molecule encoding a protein involved in the biosynthesis of a terpenoid or a precursor thereof, wherein said nucleic acid molecule is operably linked to a derepressible promoter. 1. A nucleic acid construct comprising a nucleic acid molecule encoding a protein involved in the biosynthesis of a terpenoid or a precursor thereof , wherein said nucleic acid molecule is operably linked to a derepressible promoter.2. The nucleic acid construct according to claim 1 , wherein the protein involved in the biosynthesis of a terpenoid or a precursor thereof is selected from the group consisting of geranylgeranyl diphosphate synthases or taxadiene synthases.3. The nucleic acid construct according to claim 1 , wherein the derpressible promoter is selected from the group consisting of CAT1 promoter claim 1 , FDH1 promoter claim 1 , FLD1 promoter claim 1 , PEX5 promoter claim 1 , DAK1 promoter claim 1 , FGH1 promoter claim 1 , GTH1 promoter claim 1 , G1 promoter claim 1 , G2 promoter claim 1 , G3 promoter claim 1 , G4 promoter claim 1 , G5 promoter claim 1 , G6 promoter claim 1 , FMD promoter and a functional variant thereof.4. The nucleic acid construct according to claim 1 , wherein the promoter is an orthologous promoter.5. The nucleic acid construct according to claim 1 , wherein the derepressible promoter is linked to a second promoter forming a bidirectional promoter or a bidirectional derepressible promoter.6. The nucleic acid construct according to claim 5 , wherein the second promoter is a constitutive claim 5 , derepressible or inducible promoter.7. The nucleic acid construct according to claim 6 , wherein the constitutive promoter is selected from the group consisting of a GAP promoter claim 6 , PGCW14 promoter claim 6 , TEF1 promoter claim 6 , TPI promoter claim 6 , PGK1 promoter or a histone promoter.8. The nucleic acid construct according to claim 6 , wherein the inducible promoter is ...

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Номер записи: 35
09-04-2015 дата публикации

PEST RESISTANT PLANTS

Номер: US20150101084A1
Автор: Bleeker Petronella Martina, HARING Michael Albertus, Schuurink Robert Cornelis
Принадлежит: Keygene N.V.

The disclosure provides an isolated nucleic acid molecule encoding a 7-epizingiberene synthase, a chimeric gene comprising said nucleic acid molecule, vectors comprising the same, as well as isolated 7-epizingiberene synthase proteins themselves. In addition, transgenic plants and plant cells comprising a gene encoding a 7-epizingiberene synthase, optionally integrated in its genome, and methods for making such plants and cells, are provided. Especially Solanaceae plants and plant parts (seeds, fruit, leaves, etc.) with enhanced insect pest resistance are provided. 1. An isolated protein comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence comprising at least 99% amino acid sequence identity to the amino acid sequence of SEQ ID NO:1 over the entire length.2. A recombinant nucleic acid sequence comprising a heterologous promoter operably linked to a nucleic acid sequence selected from the group consisting of:(a) a nucleic acid sequence of SEQ ID NO: 2;(b) a nucleic acid sequence that encodes a polypeptide comprising an amino acid sequence of SEQ ID NO: 1;(c) a nucleic acid sequence that is at least 99% identical to the nucleic acid sequences of (a) or (b), and encodes a 7-epizingiberene synthase;(d) a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:1 wherein at least one amino acid is substituted, deleted, inserted or added and wherein the polypeptide is functionally equivalent to the polypeptide consisting of the amino acid sequence of SEQ ID NO: 1 and(e) a nucleic acid sequence that hybridizes under stringent conditions to the nucleic acid sequences of (a), (b), or (c), and encodes a 7-epizingiberene synthase.3. An isolated nucleic acid sequence according to claim 2 , comprising the nucleotide sequence of SEQ ID NO:2.4. A vector comprising the chimeric gene according to .5. A host cell comprising the vector according to .6. A cDNA sequence comprising a nucleic acid sequence selected from the group ...

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Номер записи: 36
06-04-2017 дата публикации

Microorganisms for Biosynthesis of Limonene on Gaseous Substrates

Номер: US20170096685A1
Автор: Carr Cody A. Marcus, Jannson Christer, Reed John S.
Принадлежит: Kiverdi, Inc.

Engineered microorganisms are provided that convert gaseous substrates, such as producer gas, into limonene. In some embodiments, limonene is pumped out of the cell via an efflux pump. In some embodiments, limonene, produced as described herein, is converted through catalytic dimerization into jet fuel. Producer gas used in the processes described herein for production of limonene may be derived from sources that include gasification of waste feedstock and/or biomass residue, waste gas from industrial processes, or natural gas, biogas, or landfill gas. 1. A non-naturally occurring microorganism that is capable of growing on a gaseous substrate as a carbon and/or energy source , wherein said microorganism comprises at least one exogenous nucleic acid , and wherein said microorganism produces a terpene.2. A non-naturally occurring microorganism according to claim 1 , wherein said microorganism is a bacterial cell.3. A non-naturally occurring microorganism according to claim 1 , wherein said gaseous substrate comprises COas a carbon source.4. A non-naturally occurring microorganism according to claim 1 , wherein said gaseous substrate comprises Hand Oas an energy source.5. (canceled)6. A non-naturally occurring microorganism according to claim 1 , wherein said gaseous substrate comprises CO; Hand CO; CO claim 1 , H claim 1 , and CO; or CO and H.7. A non-naturally occurring microorganism according to claim 1 , wherein said microorganism produces the terpene when cultured in the presence of the gas substrate under conditions suitable for growth of the microorganism and production of bioproducts.8CupriavidusRalstoniaXanthobacterRhodococcusHydrogenovibrioRhodopseudomonasRhodobacterHydrogenobacterArthrobacterParacoccusMycobacteriumStreptomycesBacillus. The non-naturally occurring microorganism of claim 1 , wherein said microorganism is a sp. claim 1 , sp claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , ...

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Номер записи: 37
14-04-2016 дата публикации

EXTRACELLULAR DITERPENE PRODUCTION

Номер: US20160102331A1
Автор: BOER Viktor Marius, BROERS Nicolette Jasmijn, LAWRENCE Adam G.
Принадлежит:

The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism in a suitable fermentation medium, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol, whereby a diterpene or glycosylated diterpene is produced extracellularly in the fermentation medium; and b. recovering the diterpene or glycosylated diterpene from the fermentation medium. 1. A method for the production of a diterpene or a glycosylated diterpene , which method comprises:a. fermenting a recombinant microorganism in a suitable fermentation medium,wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol,whereby a diterpene or glycosylated diterpene is produced extracellularly in the fermentation medium; andb. recovering the diterpene or glycosylated diterpene from the fermentation medium.2. A method according to claim 1 , wherein the recombinant microorganism comprises one or more nucleotide sequences encoding a polypeptide having UDP-glucosyltransferase activity claim 1 ,whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least one of steviolmonoside, steviolbioside, stevioside or ...

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Номер записи: 38
16-04-2015 дата публикации

THREONINE SYNTHASE FROM NICOTIANA TABACUM AND METHODS AND USES THEREOF

Номер: US20150106971A1
Автор: Bovet Lucien, Sierro Nicolas
Принадлежит: PHILIP MORRIS PRODUCTS S.A.

There is disclosed a mutant, non-naturally occurring or transgenic plant cell comprising: (i) a polynucleotide comprising, consisting or consisting essentially of a sequence encoding a threonine synthase and having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 or at least 87% sequence identity to SEQ ID NO:4, or SEQ ID NO:5; (ii) a polypeptide encoded by any one of said polynucleotides set forth in (i); or (iii) a polypeptide having at least 95% sequence identity to SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8; or (iv) a construct, vector or expression vector comprising the polynucleotide as set forth in (i). 1. A mutant , non-naturally occurring or transgenic plant cell comprising:(i) a polynucleotide comprising, consisting or consisting essentially of a sequence encoding a threonine synthase and having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 or at least 87% sequence identity to SEQ ID NO:4, or SEQ ID NO:5;(ii) a polypeptide encoded by any one of said polynucleotides set forth in (i); or(iii) a polypeptide having at least 95% sequence identity to SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8; or(iv) a construct, vector or expression vector comprising the polynucleotide as set forth in (i).2. A mutant claim 1 , non-naturally occurring or transgenic plant comprising the plant cell according to .3. A method for increasing the concentration of free methionine in at least a part of a plant claim 1 , preferably a tobacco plant claim 1 , comprising the steps of: (i) a polynucleotide comprising, consisting or consisting essentially of a sequence encoding a threonine synthase and having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 or at least 87% sequence identity to SEQ ID NO:4, or SEQ ID NO:5;', '(ii) a polypeptide encoded by any one of said polynucleotides set forth in (i); or', '(iii) a polypeptide having at least 95% sequence identity to SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8;, '(a) ...

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Номер записи: 39
08-04-2021 дата публикации

Terpene synthase producing patchoulol and elemol, and preferably also pogostol

Номер: US20210102224A1
Автор: Adèle Margaretha Maria Liduina VAN HOUWELINGEN, Elena MELILLO, Georg Friedrich Lentzen, Hendrik Jan Bosch, Martinus Julius Beekwilder
Принадлежит: Isobionics BV

The invention is directed to a patchoulol synthase, to a nucleic acid encoding said patchoulol synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing patchoulol and elemol, and preferably also pogostol, and to a method of preparing a patchoulol synthase.

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Номер записи: 40
19-04-2018 дата публикации

METHOD FOR PRODUCING BETA-SANTALENE

Номер: US20180105837A1
Автор: Schalk Michel
Принадлежит: FIRMENICH SA

The present invention provides a method of producing β-santalene, said method comprising contacting at least one polypeptide with farnesyl pyrophosphate (FPP). In particular, said method may be carried out in vitro or in vivo to produce β-santalene, a very useful compound in the fields of perfumery and flavoring. The present invention also provides the amino acid sequence of a polypeptide useful in the method of the invention. A nucleic acid encoding the polypeptide of the invention and an expression vector containing said nucleic acid are also part of the present invention. A non-human host organism or a cell transformed to be used in the method of producing β-santalene is also an object of the present invention. 1. A recombinant polypeptide having a β-santalene synthase activity and comprising an amino acid sequence at least 90% identical to SEQ ID NO: 15.2. An isolated polypeptide having a β-santalene synthase activity and comprising the amino acid sequence of SEQ ID NO: 27.3. An isolated nucleic acid encoding a polypeptide comprising the amino acid sequence according to .4. An isolated nucleic acid molecule comprisinga) the cDNA sequence of SEQ ID NO: 14 or the complement thereof;b) a modified sequence of SEQ ID NO: 14, wherein the first 23 codons of SEQ ID NO: 14 were removed; or{'i': 'E. coli', 'c) the modified sequence of b) optimized for expression.'}5. An expression vector comprising{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'a) the nucleic acid molecule of ;'}b) a nucleic acid encoding a polypeptide comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 15, or SEQ ID NO: 27 and having a β-santalene synthase activity; orc) the nucleic acid of b) comprising a nucleotide sequence at least 75% identical to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 14, SEQ ID NO: 26 or the complement thereof.7. The non-human host organism or cell of claim 6 , wherein said non-human host organism or cell is a plant ...

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Номер записи: 41
02-04-2020 дата публикации

PROCESS OF PRODUCING MONOTERPENES

Номер: US20200102585A1
Автор: KARUPPIAH Vijaykumar, LEFERINK Nicole G. H., Scrutton Nigel S.
Принадлежит:

The present invention relates to a process of producing a monoterpene and/or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and/or derivatives thereof and a recombinant microorganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by expression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from 1. A process of producing a monoterpene and/or derivatives thereof in a host microorganism comprising the following steps:(a) providing a host microorganism genetically modified to express a bacterial monoterpene synthase (mTS); and(b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof.2E. coli.. The process according to wherein the host microorganism comprises3Streptomyces. The process according to wherein the bacterial mTS comprises a mTS.4Streptomyuces clavuligerus. The process according to wherein the bacterial mTS comprises a mTS.5. The process according to wherein the bacterial mTS comprises cineole-1 claim 1 ,8 synthase or linalool synthase.6. The process according to wherein the process results in a monoterpene yield of at least 100 mg/L.7. The process according to wherein the process results in a monoterpene yield of at least 300 mg/L.8. A microorganism for use in producing a monoterpene and/or derivatives thereof according to the process of .9. A recombinant microorganism adapted to conduct the following step: converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by expression of a ...

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Номер записи: 42
11-04-2019 дата публикации

MODIFIED CELL

Номер: US20190106684A1
Автор: GRAHAM Ian, King Andrew
Принадлежит: THE UNIVERSITY OF YORK

The present disclosure relates to nucleic acids that encode enzyme activities involved in the synthesis of lathyranes, intermediates in the synthesis of lathyranes and also compounds derived from lathyranes such as tiglianes, daphnanes and ingenanes; cells transformed with the nucleic acid molecules and vectors comprising the nucleic acid molecules. 1. An isolated cell transformed or transfected with an expression vector adapted to express a nucleic acid molecule comprising(a) the nucleotide sequence of SEQ ID NO: 3,(b) a nucleotide sequence comprising at least 90% sequence identity to the nucleotide sequence of SEQ ID NO: 3 and encoding a polypeptide that has casbene-9-oxidase activity,(c) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or(d) a nucleic acid sequence encoding a polypeptide that is greater than 96% identical to the amino acid sequence of SEQ ID NO: 2 and has casbene-9-oxidase activity.25.-. (canceled)6. The isolated cell according to claim 1 , wherein said isolated cell is transformed with at least one vector comprising a nucleotide molecule selected from the group consisting of:i) the nucleotide sequence of SEQ ID NO: 3, or a nucleotide sequence comprising at least 90% sequence identity to the nucleotide sequence of SEQ ID NO: 3 and encodes a polypeptide that has casbene-9-oxidase activity; andii) the nucleotide sequence of SEQ ID NO: 6, or a nucleotide sequence comprising at least 90% sequence identity to the nucleotide sequence of SEQ ID NO: 6 and encodes a polypeptide that has casbene synthase activity; andiii) the nucleotide sequence of SEQ ID NO: 4, or a nucleotide sequence comprising at least 90% sequence identity to the nucleotide sequence of SEQ ID NO: 4 and encodes a polypeptide that has casbene 5,6-oxidase activity.7. The isolated cell according to claim 1 , wherein said isolated cell transformed with at least one vector comprising a nucleotide molecule selected from the group consisting ...

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Номер записи: 43
13-05-2021 дата публикации

FUSION CONSTRUCTS AS PROTEIN OVER-EXPRESSION VECTORS

Номер: US20210139921A1
Автор: Formighieri Cinzia, Melis Anastasios
Принадлежит:

This invention provides compositions and methods for providing high product yield of transgenes expressed in cyanobacteria and microalgae. 1. An expression construct comprising a nucleic acid sequence encoding a fusion protein comprising (a) a protein of interest encoded by a transgene and (b) a polypeptide leader sequence , wherein the transgene is codon-optimized for expression in cyanobacteria , and is fused to the 3′ end of a nucleic acid sequence encoding the polypeptide leader sequence , wherein the polypeptide leader sequence is an exogenous antibiotic resistance protein that is over-expressed in cyanobacteria at a level of at least 1% of the total cellular protein.2. The expression construct of claim 1 , wherein the exogenous antibiotic resistance protein confers resistant to kanamycin claim 1 , chloramphenicol claim 1 , streptomycin claim 1 , or spectinomycin.3. The expression construct of claim 1 , wherein the transgene encodes a terpene synthase.4. The expression construct of claim 3 , wherein the terpene synthase is an isoprene synthase.5. The expression construct of claim 3 , wherein the terpene synthase is a monoterpene synthase.6. The expression construct of claim 5 , wherein the monoterpene synthase is beta-phellandrene synthase.7. The expression construct of claim 6 , wherein the beta-phellandrene synthase is lavender claim 6 , tomato claim 6 , grand fir claim 6 , pine claim 6 , or spruce beta-phellandrene synthase.8. The expression construct of claim 3 , wherein the terpene synthase is a sesquiterpene synthase.9. The expression construct of claim 8 , wherein the sesquiterpene synthase is a farnesene synthase claim 8 , a zingiberene synthase claim 8 , a caryophellene synthase claim 8 , a longifolene synthase claim 8 , a taxadiene synthase claim 8 , or a dictyophorine synthase.10. A host cell comprising an expression construct of .11. The host cell of claim 10 , wherein the host cell is a cyanobacteria host cell.12. The host cell of claim 11 , ...

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Номер записи: 44
03-05-2018 дата публикации

Heterologous Pathway to Produce Terpenes

Номер: US20180119176A1
Автор: Christopher B. Eiben, Jay D. Keasling
Принадлежит: UNIVERSITY OF CALIFORNIA

Cells comprising a heterologous metabolic pathway are configured to produce a terpene product containing non-multiples of five carbon, particularly wherein the pathway comprises heterologous Lepidoptera insect juvenile hormone biosynthetic pathway enzymes of the insect's mevalonate pathway.

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Номер записи: 45
27-05-2021 дата публикации

METHODS FOR STABILIZING PRODUCTION OF ACETYL-COENZYME A DERIVED COMPOUNDS

Номер: US20210155939A1
Автор: JIANG Hanxiao, Meadows Adam
Принадлежит:

The present disclosure relates to the use of a switch for the production of heterologous non-catabolic compounds in microbial host cells. In one aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured under aerobic conditions followed by microaerobic conditions, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. In another aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured in the presence of maltose followed by the reduction or absence of maltose, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. 1. A method for producing a heterologous non-catabolic compound in a genetically modified host cell , the method comprising:(a) culturing a population of genetically modified host cells in a culture medium comprising a carbon source under aerobic conditions, wherein the host cell comprises one or more heterologous nucleic acids encoding one or more enzymes of an enzymatic pathway for making the heterologous non-catabolic compound, wherein expression of the one or more enzymes is positively regulated by the activity of a microaerobic-responsive promoter, wherein the aerobic conditions limit the amount of heterologous non-catabolic compound produced by the host cells; and(b) culturing said population or a subpopulation thereof in a culture medium comprising a carbon source under microaerobic conditions, wherein said microaerobic conditions increases the production of the non-catabolic compound by said population or subpopulation thereof.236.-. (canceled)37. A fermentation composition comprising a population of genetically modified host cells in a culture medium comprising a carbon source , wherein the host cell comprises one or more heterologous nucleic acids ...

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Номер записи: 46
21-05-2015 дата публикации

ENGINEERING PLANTS WITH RATE LIMITING FARNESENE METABOLIC GENES

Номер: US20150141714A1
Автор: Blakeslee Joshua, Cornish Katrina, Crasta Oswald, Folkerts Otto, Jessen Dave, Nair Ramesh
Принадлежит:

The disclosed invention provides methods and compositions for increasing terpenoid production, such as sesquiterpenoids, such as farnesene, in plant cells. 1. A plant cell having increased production of at least one terpenoid native to a plant , the method comprising expressing in a plant cell a heterologous nucleic acid encoding for (a) HMG-CoA reductase , (b) 1-deoxy-D-xylulose-5-phosphate synthase , (c) farnesyl pyrophosphate synthase , and (d) β-farnesene synthase , wherein production of the at least one terpenoid is significantly increased when compared to a wild-type plant cell not encoding the heterologous nucleic acids.2. The method of claim 1 , wherein{'i': Arabidopsis, Oryza, Saccharomyces', 'Hevea, 'a. the HMG-CoA reductase is an , or HMG-CoA reductase;'}{'i': Arabidopsis, Oryza, Saccharomyces', 'Zea, 'b. the 1-deoxy-D-xyululose-5-phophate is an , or 1-deoxy-D-xyululose;'}{'i': Arabidopsis, Oryza', 'Solanum, 'c. the farnesyl pyrophosphate synthase is an , or farnesyl pyrophosphate; or'}{'i': Arabidopsis, Oryza', 'Artemisia, 'd. the β-farnesene synthase is an , or β-farnesene synthase.'}3. The method of claim 2 , wherein{'i': Arabidopsis thaliana, Oryza sativa, Saccharomyces cerevisiae', 'Hevea, 'a. the HMG-CoA reductase is an , or HMG-CoA reductase;'}{'i': Arabidopsis thaliana, Oryza sativa, Saccharomyces cerevisiae', 'Zea mays, 'b. the 1-deoxy-D-xyululose-5-phophate is an , or 1-deoxy-D-xyululose;'}{'i': Arabidopsis thaliana, Oryza sativa', 'Solanum lycopersicon, 'c. the farnesyl pyrophosphate synthase is an , or farnesyl pyrophosphate;'}{'i': Arabidopsis thaliana, Oryza sativa', 'Artemisia annua β, 'd. the β-farnesene synthase is an , or -farnesene synthase.'}4. The method of claim 3 , wherein at least one nucleic acid is codon-optimized for expression in a plant.5. The method of claim 3 , whereina. the HMG-CoA reductase is encoded by a polynucleotide having at least 70% sequence identity to a nucleic acid sequence selected from the group consisting of ...

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Номер записи: 47
09-05-2019 дата публикации

VALENCENE SYNTHASE

Номер: US20190136221A1
Автор: DE JONG Rene M., SONKE Theodorus
Принадлежит:

The present invention relates to a novel valencene synthase, to a nucleic acid encoding such valencene synthase, to a host cell comprising said encoding nucleic acid sequence and to a method or preparing valencene, comprising converting farnesyl diphosphate to valencene in the presence of a valencene synthase according to the invention. 1. Valencene synthase , which valence synthase has an increased productivity towards the conversion of farnesyl diphosphate into valencene (expressed as molar amount of valencene formed per hour) compared to a valencene synthase represented by SEQ ID NO: 2 , or which valencene synthase comprises an amino acid sequence represented by SEQ ID NO: 3 provided that at least one position marked ‘X’ in SEQ ID NO: 3 is different from the corresponding position in SEQ ID NO: 2.2. Valencene synthase according to claim 1 , wherein the valencene synthase has an increased specific productivity claim 1 , increased stability claim 1 , increased product specificity (relative to the conversion of farnesyl diphosphate into Germacrene A) or an increased expression in a host cell claim 1 , compared to a valencene synthase represented by SEQ ID NO: 2.3. Valencene synthase according to claim 2 , wherein the product specificity claim 2 , expressed as the molar ratio valencene formed from farnesyl diphospate to Germacrene A formed from farnesyl diphosphate (under test conditions) claim 2 , is 10 or more claim 2 , preferably 13-30 claim 2 , in particular 15-25.4. Valencene synthase according to claim 1 , wherein the specific productivity of the valencene synthase claim 1 , expressed as the molar amount of valencene formed per hour per amount of valencene synthase is at least 1.5 times claim 1 , preferably 2.0 to 10 times claim 1 , in particular 2.5 to 5 times the specific productivity of the valencene synthase represented by SEQ ID NO: 2.5. Valencence synthase according to claim 1 , wherein the valencene synthase has at least one modification claim 1 , in ...

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Номер записи: 48
24-05-2018 дата публикации

Microbial fermentation for the production of terpenes

Номер: US20180142265A1
Автор: Fungmin Liew, Michael Koepke, Wendy Yiting Chen
Принадлежит: Lanzatech New Zealand Ltd

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.

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Номер записи: 49
25-05-2017 дата публикации

METHODS, HOSTS, AND REAGENTS RELATED THERETO FOR PRODUCTION OF UNSATURATED PENTAHYDROCARBONS, DERIVATIVES AND INTERMEDIATES THEREOF

Номер: US20170145441A1
Автор: Conradie Alex Van Eck
Принадлежит:

This application describes methods, including non-naturally occurring methods, for biosynthesizing unsaturated pentahydrocarbons, such as isoprene and intermediates thereof, via the mevalonate pathway, as well as non-naturally occurring hosts for producing isoprene. 1. A method for synthesizing isoprene in a chemolithotrophic host comprising:enzymatically converting acetoacetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA using a polypeptide having the activity of a hydroxymethylglutaryl-CoA synthase enzyme having the amino acid sequence set forth in SEQ ID No: 2 or 10 or a functional fragment of said enzyme; andenzymatically converting 3-hydroxy-3-methylglutaryl-CoA to (R)-mevalonate using a polypeptide having the activity of a hydroxymethylglutaryl Co-A reductase enzyme having the amino acid sequence set forth in SEQ ID No: 3 or 9 or a functional fragment of said enzyme.2. (canceled)3. The method of claim 1 , further comprising at least one of:enzymatically converting acetyl-CoA to acetoacetyl-CoA using a polypeptide having the activity of an acetyl-CoA acetyltransferase enzyme having the amino acid sequence set forth in SEQ ID No: 1 or 9 or a functional fragment of said enzyme;enzymatically converting (R)-mevalonate to (R)-5-phosphomevalonate using a polypeptide having the activity of a mevalonate-kinase enzyme having the amino acid sequence set forth in SEQ ID No: 4 or 11 or a functional fragment of said enzyme;enzymatically converting (R)-5-phosphomevalonate to (R)-5-diphosphomevalonate using a polypeptide having the activity of a phosphomevalonate kinase enzyme having the amino acid sequence set forth in SEQ ID No: 5 or 12 or a functional fragment of said enzyme;enzymatically converting (R)-5-diphosphomevalonate to isopentenyl diphosphate using a polypeptide having the activity of a diphosphomevalonate decarboxylase enzyme having the amino acid sequence set forth in SEQ ID No: 6 or 13 or a functional fragment of said enzyme;enzymatically converting isopentenyl ...

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Номер записи: 50
02-06-2016 дата публикации

MICROORGANISMS AND METHODS FOR THE BIOSYNTHESIS OF (2-HYDROXY-3METHYL-4-OXOBUTOXY) PHOSPHONATE

Номер: US20160152989A1
Автор: Osterhout Robin E.
Принадлежит:

The invention provides non-naturally occurring microbial organisms having a (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate pathway, p-toluate pathway, and/or terephthalate pathway. The invention additionally provides methods of using such organisms to produce (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate pathway, p-toluate pathway or terephthalate pathway. 1. A non-naturally occurring microbial organism having carbon monoxide dehydrogenase or hydrogenase activity and a reductive tricarboxylic acid (rTCA) pathway , wherein the microbial organism comprises (i) at least one exogenous nucleic acid encoding a rTCA pathway enzyme expressed in a sufficient amount to convert CO , COor Hto acetyl-CoA , wherein the rTCA enzyme is selected from the group consisting of an ATP citrate-lyase , a citrate lyase , an aconitase , an isocitrate dehydrogenase , an alpha-ketoglutarate:ferredoxin oxidoreductase , a succinyl-CoA synthetase , a succinyl-CoA transferase , a fumarate reductase , a fumarase , and a malate dehydrogenase; and (ii) at least one exogenous nucleic acid encoding a biosynthetic product pathway enzyme expressed in a sufficient amount to produce the biosynthetic product; and wherein the microbial organism requires a carbon source as an energy source.2. The non-naturally occurring microbial organism of claim 1 , wherein the biosynthetic product is p-toluate claim 1 , terephthalate or (2-hydroxy-3-methyl-4-oxobutoxy)phosphonate.3. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism does not require light for growth.4. The non-naturally occurring microbial organism of further comprises a ferredoxin.5. The non-naturally occurring microbial organism of further comprises a reducing equivalent producing pathway comprising at least one exogenous nucleic acid encoding a reducing equivalent producing enzyme selected from the group consisting of a carbon monoxide dehydrogenase claim 1 , a hydrogenase and a NAD(P)H:ferredoxin oxidoreductase.6. ...

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Номер записи: 51
16-05-2019 дата публикации

Genetically Modified Microorganisms

Номер: US20190144814A1
Автор: Frank Stefanie, Lee Matthew John, Warren Martin James
Принадлежит: University of Kent

The present invention relates to genetically modified microorganisms comprising one or more heterologous nucleic acid molecules together encoding at least three different proteins, each protein comprising an enzymatic domain and a bacterial microcompartment-targeting signal polypeptide, wherein said enzymatic domains each catalyse a different substrate to product conversion in the same metabolic pathway, and wherein said microorganisms are essentially free of bacterial microcompartments (BMCs); and to cell free systems comprising aggregates comprising at least three different proteins, each protein comprising an enzymatic domain and a bacterial microcompartment-targeting signal polypeptide, wherein said enzymatic domains each catalyse a different substrate to product conversion in the same metabolic pathway, and wherein said system does not comprise bacterial microcompartments; and to methods for the production of said microorganisms and cell free systems and their use in methods of producing a product of interest. 1. A genetically modified microorganism comprising wherein said enzymatic domains each catalyse a different substrate to product conversion in the same metabolic pathway, and', 'wherein said microorganism is essentially free of bacterial microcompartments (BMCs)., 'one or more heterologous nucleic acid molecules together encoding at least three different proteins, each protein comprising an enzymatic domain and a bacterial microcompartment-targeting signal polypeptide,'}2. The genetically modified microorganism of claim 1 , wherein said microorganism comprises one or more heterologous nucleic acid molecules together encoding at least four different proteins claim 1 , each protein comprising an enzymatic domain and a bacterial microcompartment-targeting signal polypeptide claim 1 ,wherein said enzymatic domains each catalyse a different substrate to product conversion in the same metabolic pathway.3. The genetically modified microorganism of claim 1 , ...

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Номер записи: 52
16-05-2019 дата публикации

Microorganisms for Biosynthesis of Limonene on Gaseous Substrates

Номер: US20190144891A1
Автор: Carr Cody A. Marcus, Jansson Christer, Reed John S.
Принадлежит: Kiverdi, Inc.

Engineered microorganisms are provided that convert gaseous substrates, such as producer gas, into limonene. In some embodiments, limonene is pumped out of the cell via an efflux pump. In some embodiments, limonene, produced as described herein, is converted through catalytic dimerization into jet fuel. Producer gas used in the processes described herein for production of limonene may be derived from sources that include gasification of waste feedstock and/or biomass residue, waste gas from industrial processes, or natural gas, biogas, or landfill gas. 136.-. (canceled)37. A composition comprising a non-naturally occurring microorganism that is capable of growing on a gaseous substrate as a carbon and energy source , and a culture medium in contact with a gaseous substrate that comprises an inorganic gaseous electron donor and an inorganic gaseous electron acceptor ,wherein the microorganism comprises at least one exogenous nucleic acid, andwherein the gaseous substrate is utilized by the microorganism for production of a terpene in the culture medium using a combination of the electron donor and the electron acceptor as an energy source.38. The composition of claim 37 , wherein the gaseous substrate comprises CO claim 37 , CO claim 37 , and/or CHas a carbon source.39. The composition of claim 37 , wherein the microorganism is a knallgas microorganism.40. The composition of claim 39 , wherein the gaseous substrate comprises COas a carbon source claim 39 , Has an electron donor claim 39 , and Oas an electron acceptor.41. The composition of claim 39 , wherein the gaseous substrate comprises Hand Oas an energy source.42. The composition of claim 37 , wherein the microorganism comprises at least one exogenous nucleic acid encoding an efflux pump and/or comprises the ability to overexpress a native efflux pump.43A. borkumensisE. coli. The composition of claim 42 , wherein the exogenous nucleic acid encoding an efflux pump encodes an efflux pump from or AcrB protein.44. ...

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Номер записи: 53
07-05-2020 дата публикации

Production of Steviol Glycosides in Microorganisms

Номер: US20200140912A1
Автор: Blom Charlotte, Hansen Esben Halkjaer, Hansen Jorgen, HICKS Paula M., Houghton-Larsen Jens, Kishore Ganesh M., Mikkelsen Michael Dalgaard, Motion Michael, Tavares Sabina
Принадлежит:

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements. 1. A method for producing Rebaudioside D (RebD) , Rebaudioside E (RebE) , or a mixture thereof , comprising contacting a precursor steviol glycoside having a 13-O-glucose , a 19-O-glucose , or both the 13-O-glucose and the 19-O-glucose with a polypeptide capable of beta 1 ,2 glycosylation of the C-2′ of the 13-O-glucose , 19-O-glucose , or both the 13-O-glucose and the 19-O-glucose of the precursor steviol glycoside and a UDP-glucose in a reaction mixture under suitable conditions for the transfer of one or more glucose moiety to the C2′ of the 13-O-glucose , 19-O-glucose or both 13-O-glucose and 19-O-glucose in the precursor steviol glycoside; thereby producing RebD , RebE , or a mixture thereof.2. The method of claim 1 , comprising further contacting the reaction mixture with:(a) a polypeptide capable of glycosylating a precursor steviol glycoside having a C-13 hydroxyl group present in the reaction mixture at its C-13 hydroxyl group; and/or(b) a polypeptide capable of glycosylating a precursor steviol glycoside having a C-19 carboxyl group present in the reaction mixture at its C-19 carboxyl group; and/or(c) a polypeptide capable of beta 1,3 glycosylation of the C3′ of the 13-O-glucose, of the 19-O-glucose or both the 13-O-glucose and the 19-O-glucose of the precursor steviol glycoside having a 13-O-glucose, a 19-O-glucose, or both the 13-O-glucose and the 19-O-glucose present in the reaction mixture.3. The method of claim 1 , which is an in vitro method comprising supplying the UDP-glucose or a cell-free system for regeneration of the UDP-glucose.4. The method of ...

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Номер записи: 54
09-06-2016 дата публикации

DITERPENE PRODUCTION IN YARROWIA

Номер: US20160160257A1
Автор: BOER Viktor Marius, BROERS Nicolette Jasmijn, LAWRENCE Adam G.
Принадлежит:

The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism of the genus in a suitable fermentation medium at a temperature of about 29° C. or higher, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol; and b. recovering the diterpene or glycosylated diterpene. 1. A method for the production of a diterpene or a glycosylated diterpene , which method comprises:{'i': 'Yarrowia', 'a. fermenting a recombinant microorganism of the genus in a suitable fermentation medium at a temperature of about 29° C. or higher,'}wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol; andb. recovering the diterpene or glycosylated diterpene.2. A process according to claim 1 , wherein the recombinant microorganism comprises one or more nucleotide sequences encoding a polypeptide having UDP-glucosyltransferase activity claim 1 ,whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least one of steviolmonoside, steviolbioside, stevioside or rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside M, rubusoside, dulcoside A.3 ...

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Номер записи: 55
18-06-2015 дата публикации

TERPENE SYNTHASES FROM SANTALUM

Номер: US20150167021A1
Автор: Bohlmann Joerg, Jones Christopher, Moniodis Jessie, Zulak Katherine
Принадлежит:

An isolated nucleic acid molecule that encodes a terpene synthase and is selected from among: a) a nucleic acid molecule comprising the sequence of nucleotides set forth in SEQ ID NO: 1, SEQ ID NO: 3 or SEQ ID NO: 5; b) a nucleic acid molecule that is a fragment of (a); c) a nucleic acid molecule comprising a sequence of nucleotides that is complementary to (a) or (b); and d) a nucleic acid molecule that encodes a terpene synthase having at least or at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity to any one of (a)-(c); wherein the nucleic acid molecule encodes a terpene synthase. 1Santalum. A host cell , comprising a nucleic acid molecule that encodes a species terpene synthase that comprises:(i) the sequence of amino acid resides set forth in SEQ ID NO:2, 4 or 6 or a catalytically active fragment thereof; or(ii) a sequence of amino acid residues that has at least 95% or greater sequence identity to the sequence of amino acid residues set forth in SEQ ID NO: 2, or a catalytically active fragment thereof, wherein:the nucleic acid molecule is heterologous to the cell; andthe encoded synthase catalyzes the production of an α-santalene, α-trans-bergamotene, epi-β-santalene and β-santalene concurrently.2. The host cell of claim 1 , wherein the encoded synthase comprises a serine in place of the proline at residue 143 of SEQ ID NO: 2.3Santalum. A vector claim 1 , comprising a nucleic acid molecule that encodes a species terpene synthase that comprises:(i) the sequence of amino acid resides set forth in SEQ ID NO:2, 4 or 6 or a catalytically active fragment thereof; or(ii) a sequence of amino acid residues that has at least 95% or greater sequence identity to the sequence of amino acid residues set forth in SEQ ID NO: 2, or a catalytically active fragment thereof,wherein the encoded synthase catalyzes the production of an α-santalene, α-trans-bergamotene, epi-β-santalene and β-santalene concurrently.4. The vector of claim 3 , wherein the ...

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Номер записи: 56
23-06-2016 дата публикации

METHOD FOR PRODUCING ALPHA-SANTALENE

Номер: US20160177288A1
Автор: Schalk Michel
Принадлежит:

The present invention provides a method of producing α-santalene by contacting at least one polypeptide with farnesyl phyrophosphate (fpp). In particular, the method may be carried out in vitro or in vivo to produce α-santalene, a very useful compound in the fields of perfumery and flavoring. The present invention also provides the amino acid sequence of a polypeptide useful in the method of the invention. A nucleic acid encoding the polypeptide of the invention and an expression vector containing the nucleic acid represent part of the present invention. A non-human host organism and a cell transformed to be used in the method of producing α santalene are also part of the present invention. 1. A method for producing α-santalene comprising:a) contacting FPP with at least one polypeptide having an α-santalene synthase activity and comprising an amino acid sequence at least 50% identical to SEQ ID NO:1;b) optionally, isolating the α-santalene produced in step a).2. The method of claim 1 , wherein step a) comprises cultivating a non-human host organism or cell capable of producing FPP and transformed to express at least one polypeptide comprising an amino acid sequence at least 50% identical to SEQ ID NO:1 and having an α-santalene synthase activity claim 1 , under conditions conducive to the production of α-santalene.3. The method of claim 2 , wherein the method further comprises claim 2 , prior to step a) claim 2 , transforming a non human host organism or cell capable of producing FPP with at least one nucleic acid encoding a polypeptide comprising an amino acid sequence at least 50% identical to SEQ ID NO:1 and having an α-santalene synthase activity claim 2 , so that said organism expresses said polypeptide.4. The method of claim 3 , wherein the at least one nucleic acid encoding the α-santalene synthase comprises a nucleotide sequence at least 50% claim 3 , at least 70% claim 3 , or at least 90% identical to SEQ ID NO:2 claim 3 , the nucleotide sequence SEQ ID NO: ...

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Номер записи: 57
21-06-2018 дата публикации

Fusion constructs as protein over-expression vectors

Номер: US20180171342A1
Автор: Anastasios Melis, Cinzia FORMIGHIERI
Принадлежит: UNIVERSITY OF CALIFORNIA

This invention provides compositions and methods for providing high product yield of transgenes expressed in cyanobacteria and microalgae.

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Номер записи: 58
21-06-2018 дата публикации

PRODUCTION OF BETA-PHELLANDRENE USING GENETICALLY ENGINEERED PHOTOSYNTHETIC MICROORGANISMS

Номер: US20180171363A1
Автор: DAVIES Fiona K., Melis Anastasios, Wintz Hsu-Ching Chen, ZURBRIGGEN Andreas
Принадлежит:

The present invention provides methods and compositions for producing β-phellandrene hydrocarbons from a photosynthetic microorganism such as cyanobacteria. 1. A method of obtaining β-phellandrene from photosynthetic microorganisms , the method comprising:culturing a strain of a photosynthetic microorganism that has been genetically modified to express a heterologous β-phellandrene synthase under conditions in which the β-phellandrene synthase is expressed; andcollecting from the surface of the culture medium β-phellandrene hydrocarbons that have spontaneously diffused into the medium from the photosynthetic microorganisms across the cell wall, wherein the culture is in a continuous growth phase and the β-phellandrene hydrocarbons are continuously generated.2. The method of claim 1 , wherein the strain of the photosynthetic microorganism is a cyanobacteria strain or a microalgae strain.3. The method of claim 1 , wherein the strain of the photosynthetic microorganism is a cyanobacteria strain.4Synechocystis, Synechococcus, Arthrospira, NostocAnabaena.. The method of claim 3 , wherein the cyanobacteria strain is from a genus selected from the group consisting of claim 3 , and5. The method of claim 3 , where the β-phellandrene synthase is encoded by a nucleic acid having at least 80% nucleic acid sequence identity to SEQ ID NO:3.6. The method of claim 1 , wherein the strain of the photosynthetic microorganism is a microalgae strain.7. The method of claim 1 , wherein the β-phellandrene synthase has at least 70% identity to SEQ ID NO:1.8. The method of claim 7 , wherein the β-phellandrene synthase has at least 90% identity to SEQ ID NO:1.9. The method of claim 1 , wherein collecting β-phellandrene hydrocarbons comprises siphoning or skimming the β-phellandrene hydrocarbons from the surface of the culture medium.10. The method of claim 1 , wherein collecting β-phellandrene hydrocarbons comprises overlaying a solvent onto the surface of the culture medium.11. The method of ...

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Номер записи: 59
22-06-2017 дата публикации

PRODUCTION OF ISOPRENE BY METHANE-PRODUCING ARCHAEA

Номер: US20170175145A1
Автор: Aldridge Jared Thomas, Buan Murphy Nicole Roswitha, Carr Sean Robert, Weber Karrie A.
Принадлежит:

Plasmid vectors and use of plasmid vectors in methods for producing methane and isoprene using are disclosed. Particularly, plasmid vectors that express isoprene synthase (ispS) are prepared and inserted into methanogens, such as , to allow for co-production of methane and isoprene. In one embodiment, the methods of the present disclosure can be used for wastewater management. 1. A host cell comprising a vector comprising a nucleic acid encoding isoprene synthase.2. The host cell of wherein the cell is a microorganism selected from the group consisting of methanogen claim 1 , methanotroph claim 1 , and acetogen.3Methanosarcinales. The host cell of wherein the cell is a strain.4Methanosarcinales acetivoransMethanosarcinales barkeri.. The host cell of wherein the cell is selected from the group consisting of and5. The host cell of wherein the vector comprises a nucleic acid having a nucleotide sequence at least 90% identity to SEQ ID NO:1.6. The host cell of wherein the vector comprises a nucleic acid having the nucleotide sequence of SEQ ID NO:1.7. The host cell of wherein the vector comprises a nucleic acid having a nucleotide sequence at least 90% identity to SEQ ID NO:2.8. A vector comprising a nucleic acid encoding isoprene synthase.9. The vector of comprising a nucleic acid having a nucleotide sequence at least 90% identity to SEQ ID NO:1.10. The vector of comprising the nucleic acid having a nucleotide sequence having at least 95% identity to SEQ ID NO:111. The vector of comprising the nucleic acid having a nucleotide sequence of SEQ ID NO:1.12. The vector of comprising a nucleic acid having a nucleotide sequence at least 90% identity to SEQ ID NO:2.13. A method of preparing a microbial strain capable of co-producing methane and isoprene claim 8 , the method comprising:preparing a vector comprising a nucleic acid encoding isoprene synthase;introducing the vector into a host cell; andculturing the host cell including the vector.14Populus alba.. The method of ...

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Номер записи: 60
08-07-2021 дата публикации

GENETICALLY ENCODED SYSTEM FOR CONSTRUCTING AND DETECTING BIOLOGICALLY ACTIVE AGENTS

Номер: US20210206813A1
Автор: Fox Jerome, Hongdusit Akarawin, Kim Edward, Sarkar Ankur
Принадлежит: The Regents of the University of Colorado, a body corporate

This invention relates to the field of genetic engineering. Specifically, the invention relates to the construction of operons to produce biologically active agents. For example, operons may be constructed to produce agents that control the function of biochemical pathway proteins (e.g., protein phosphatases, kinases and/or proteases). Such agents may include inhibitors and modulators that may be used in studying or controlling phosphatase function associated with abnormalities in a phosphatase pathway or expression level. Fusion proteins, such as light activated protein phosphatases, may be genetically encoded and expressed as photoswitchable phosphatases. Systems are provided for use in controlling phosphatase function within living cells or in identifying small molecule inhibitors/activator/modulator molecules of protein phosphatases associated with cell signaling. 1. A method of using a genetically encoded detection operon system , comprising , 'i. an inhibitor detection operon, comprising', 'a. providing,'} 1. a region of DNA encoding a first promoter;', '2. a first gene encoding a first fusion protein comprising a substrate recognition domain linked to a DNA-binding protein;', '3. a second gene encoding a second fusion protein comprising a substrate domain linked to a protein capable of recruiting RNA polymerase to DNA;', '4. a region of DNA encoding a second promoter;', '5. a third gene for a protein kinase;', '6. a fourth gene for a molecular chaperone;', '7. a fifth gene for a protein phosphatase;, 'Part A: a first region of DNA in operable combination comprising 8. a first DNA sequence encoding an operator for said DNA-binding protein;', '9. a second DNA sequence encoding a binding site for RNA polymerase; and', '10. at least one first gene of interest (GOI);', 'ii. a mevalonate-dependent isoprenoid pathway operon not containing a terpene synthase gene, under control of a fourth promoter,', 'iii. a third DNA sequence under control of a fifth promoter ...

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Номер записи: 61
09-07-2015 дата публикации

ISOPRENE SYNTHASE VARIANTS FOR IMPROVED MICROBIAL PRODUCTION OF ISOPRENE

Номер: US20150191712A1
Автор: Bott Richard R., Cervin Marguerite A., JR. James T., Kellis, McAuliffe Joseph C., Miasnikov Andrei, PERES Caroline M., RIFE Christopher L., WELLS Derek H., Weyler Walter, Whited Gregory M.
Принадлежит:

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers. 18-. (canceled)9P. alba. An isolated isoprene synthase variant , wherein the variant comprises one or more amino acid substitution(s) at one or more amino acid residue(s) of a wild type isoprene synthase , wherein said one or more amino acid residue(s) correspond to amino acid residues of isoprene synthase having the sequence of SEQ ID NO:120 , wherein said amino acid substitution(s) are selected from the group consisting of V10M , F12S , T15A , E18G , V58I , V58F , L70Q , L70R , L70V , L70T , T71P , V79L , E89D , G94A , S119F , F120L , G127R , E175V , T212I , S257A , R262G , A266G , F280L , N297K , F305L , L319M , E323K , A328T , D342E , A359T , K366N , E368D , L374M , S396T , V418S , K438N , H440R , T442A , I449V , A469S , K500R , K505Q , G507S , S509N , F511Y , and N532K; and wherein the isoprene synthase variant has increased isoprene synthase activity compared to the wild type isoprene synthase.10. The isolated isoprene synthase variant of claim 9 , wherein increased isoprene synthase activity is indicated by a host cell comprising the isoprene variant growing at a faster rate in the presence of dimethylallyl pyrophosphate (DMAPP) compared to a host cell comprising the wild type isoprene synthase.11. The isolated isoprene synthase variant of claim 9 , wherein the isoprene synthase is a poplar isoprene synthase variant.12P. alba. The isolated poplar isoprene synthase variant of claim 11 , wherein the wild type isoprene synthase is isoprene synthase of SEQ ID NO:120.13. The isolated poplar isoprene synthase variant of claim 11 , comprising the L70R substitution ...

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Номер записи: 62
07-07-2016 дата публикации

Method for producing terpenes

Номер: US20160194674A1
Автор: Bromann Kirsi, Nakari-Setala Tiina, Ruohonen Laura, Toivari Mervi
Принадлежит: TEKNOLOGIAN TUTKIMUSKESKUS VTT

The present invention concerns a method for producing terpenes in fungi comprising the steps of (a) providing a modified terpene biosynthetic gene cluster inside a host cell, wherein one or more of the naturally occurring genes or promoters of the cluster have been replaced, truncated or removed, (b) providing a transcription factor inside the host cell, the transcription factor activating the terpene biosynthetic gene cluster; (c) cultivating said host in conditions allowing the expression of the transcription factor activating the cluster; and optionally (d) recovering the thus produced terpene product. 1. Method for producing terpenes in fungi comprising the steps of:providing a modified terpene biosynthetic gene cluster inside a host cell, the gene cluster having naturally occurring terpene biosynthetic genes and promoters, wherein one or more of these genes or promoters have been replaced, truncated or removed,providing a transcription factor inside the host cell so that the transcription factor is operably linked to one of the natural or modified promoters, the transcription factor activating the terpene biosynthetic gene cluster having terpene biosynthetic genes and regulatory regions operably linked to said genes; andcultivating said host in conditions allowing the expression of the transcription factor activating the cluster.2. The method of claim 1 , wherein terpene is selected from: terpenoid claim 1 , γ-terpinene claim 1 , limonene claim 1 , cymene claim 1 , cineol claim 1 , α-farnesene claim 1 , amorphadiene claim 1 , cadinene claim 1 , a caryophyllene claim 1 , a bisabolene claim 1 , a taxadiene claim 1 , a kaurene claim 1 , a fusicoccadiene claim 1 , a casbene claim 1 , or an abietadiene.3. (canceled)4. (canceled)5. (canceled)6. The method of claim 1 , wherein the host cell carries the terpene biosynthetic gene cluster having terpene biosynthetic genes.7. The method of claim 1 , wherein the terpene biosynthetic gene cluster having terpene biosynthetic ...

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Номер записи: 63
11-06-2020 дата публикации

CONVERSION OF METHYLGLYOXAL INTO HYDROXYACETONE USING NOVEL ENZYMES AND APPLICATIONS THEREOF

Номер: US20200181652A1
Автор: CORRE Gwenaëlle, Dumon-Seignovert Laurence, FAURE Laetitia, Raynaud Céline
Принадлежит: METABOLIC EXPLORER

The present invention relates to new methylglyoxal reductase (MGR) enzymes which are useful for efficiently converting methylglyoxal into hydroxyacetone. The invention more particularly relates to a method for efficiently converting methylglyoxal into hydroxyacetone using said enzymes, to a method for producing 1,2-propanediol using a microorganism overexpressing said enzymes, and to said microorganism. 115.-. (canceled)16. A method for the fermentative conversion of methylglyoxal into hydroxyacetone , comprising the step of expressing , in a microorganism , at least one methylglyoxal reductase having a catalytic efficiency k/Km equal or superior to 5 mMswherein said methylglyoxal reductase is selected from the group consisting of the YjgB enzyme of sequence SEQ ID NO: 1 and its mutants.17. The method according to claim 16 , wherein said methylglyoxal reductase is the YjgB enzyme of sequence SEQ ID NO: 1.18. The method according to claim 17 , wherein the YjgB enzyme is expressed in combination with the YahK enzyme of sequence SEQ ID NO: 3 claim 17 , the YhdN enzyme of sequence SEQ ID NO: 5 claim 17 , the Gld enzyme of sequence SEQ ID NO: 7 claim 17 , the YafB enzyme of sequence SEQ ID NO: 11 or the YqhD enzyme of sequence SEQ ID NO: 13.20. The method according to claim 19 , further comprising the step c) of purifying the 1 claim 19 ,2-propanediol recovered from step b).21. The method according to claim 19 , wherein said microorganism further comprises the deletion of the yqhD or yqhD* gene coding for the methylglyoxal reductase of sequence SEQ ID NO: 13 or SEQ ID NO: 15.22. The method according to claim 19 , wherein said microorganism further overexpresses the gldA gene coding for the NADH dependent glycerol dehydrogenase of sequence SEQ ID NO: 21 claim 19 , or a mutant thereof coding for a NADH dependent glycerol dehydrogenase of sequence SEQ ID NO: 23.23. The method according to claim 19 , wherein said microorganism further overexpresses at least one gene coding ...

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Номер записи: 64
22-07-2021 дата публикации

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

Номер: US20210222211A1
Автор: HIRAGA Kazumi, INUI MASAYUKI, KUBOTA Takeshi, SUDA Masako
Принадлежит: RESEARCH INSTITUTE OF INNOVATIVE TECHNOLOGY FOR THE EARTH

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 ...

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Номер записи: 65
23-07-2015 дата публикации

INCREASED ISOPRENE PRODUCTION USING THE ARCHAEAL LOWER MEVALONATE PATHWAY

Номер: US20150203873A1
Автор: Beck Zachary Q., CALABRIA Anthony R., Miller Michael C., NIELSEN Alex T., VAVILINE Dmitrii V.
Принадлежит:

The invention features methods for producing isoprene from cultured cells using a feedback-resistant mevalonate kinase polypeptide, such as an archaeal mevalonate kinase polypeptide. The resulting isoprene compositions may have increased yields and/or purity of isoprene. 1. A method of increasing the rate or flux of production of 3 ,3-dimethylallyl diphosphate (DMAPP) , isopentenyl diphosphate (IPP) , or a product derived from DMAPP or IPP comprising:(a) culturing cells comprising (i) one or more heterologous nucleic acids encoding feedback-resistant mevalonate kinase polypeptides, and (ii) one or more nucleic acids encoding a mevalonate (MVA) pathway polypeptide, wherein the cells are cultured under suitable culture conditions for increasing the rate or flux of production of DMAPP, IPP, or a product derived from DMAPP or IPP, and(b) producing DMAPP, IPP, or a product derived from DMAPP and/or IPP.2. The method of claim 1 , wherein the cells further comprise one or more nucleic acids encoding an isoprene synthase.3. The method of claim 2 , wherein the product derived from DMAPP and/or IPP is isoprene.4. The method of claim 1 , wherein the product derived from DMAPP and/or IPP is an isoprenoid.5. The method of claim 1 , wherein the feedback-resistant mevalonate kinase polypeptide is an archaeal mevalonate kinase.6M. mazei. The method of claim 5 , wherein the archaeal mevalonate kinase polypeptide is an mevalonate kinase.7. The method of claim 1 , wherein the one or more nucleic acids encoding an MVA pathway polypeptide is heterologous.8. The method of claim 1 , wherein the one or more nucleic acids encoding an MVA pathway polypeptide is endogenous.9. The method of claim 3 , wherein the cells produce greater than about 400 nmole/g/hr of isoprene.10. The method of claim 1 , wherein the cells are gram-positive bacterial cells claim 1 , gram-negative bacterial cells claim 1 , fungal cells claim 1 , filamentous fungal cells claim 1 , or yeast cells. ...

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Номер записи: 66
13-07-2017 дата публикации

Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene

Номер: US20170198311A1
Автор: Andrew C. Eliot, Caroline M. Peres, Dmitrii V. Vaviline, Zachary Q. Beck
Принадлежит: Goodyear Tire and Rubber Co

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes.

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Номер записи: 67
06-08-2015 дата публикации

RECOMBINANT CELL, AND METHOD FOR PRODUCING BETA-PHELLANDRENE

Номер: US20150218589A1
Автор: FURUTANI Masahiro, Iwasa Koichiro, KORI Yasuyuki, Shimoyama Takefumi, Takahashi Seiji, Uenishi Akihiro
Принадлежит:

To provide a series of techniques for obtaining β-phellandrene with high purity and in a large quantity. 1. A recombinant cell capable of producing β-phellandrene , prepared by introducing at least one nucleic acid selected from the group consisting of a nucleic acid encoding geranyl pyrophosphate synthase and a nucleic acid encoding neryl pyrophosphate synthase , and a nucleic acid encoding β-phellandrene synthase into a host cell in such a manner that these nucleic acids are expressed in the host cell.2. The recombinant cell according to claim 1 , wherein the host cell does not have methane monooxygenase.3Escherichia coli. The recombinant cell according to claim 2 , wherein the host cell is or yeast.4. The recombinant cell according to claim 1 , wherein 10 mg or more of β-phellandrene can be produced per 1 g of wet cells of the recombinant cell.5. The recombinant cell according to claim 1 , wherein the nucleic acid encoding geranyl pyrophosphate synthase encodes the following protein (a) claim 1 , (b) or (c):(a) protein consisting of an amino acid sequence represented by SEQ ID NO: 2,(b) protein consisting of an amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 2, and having geranyl pyrophosphate synthase activity, or(c) protein consisting of an amino acid sequence having a homology of 60% or more with the amino acid sequence represented by SEQ ID NO: 2, and having geranyl pyrophosphate synthase activity.6. The recombinant cell according to claim 1 , wherein the nucleic acid encoding neryl pyrophosphate synthase encodes the following protein (d) claim 1 , (e) or (f):(d) protein consisting of an amino acid sequence represented by SEQ ID NO: 4,(e) protein consisting of an amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 4, and having neryl pyrophosphate synthase activity, or(f) protein consisting ...

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Номер записи: 68
06-08-2015 дата публикации

ENHANCED PRODUCTION OF ISOPRENE USING HOST CELLS HAVING DECREASED ISPA ACTIVITY

Номер: US20150218590A1
Автор: McAuliffe Joseph C., MUIR Rachel E., NIELSEN Alex T., PERES Caroline M., VAVILINE Dmitrii V., WELLS Derek H.
Принадлежит:

This invention relates to recombinant microorganisms capable of producing isoprene and isoprene production with the use of such recombinant microorganism with good efficiency. In this invention, functional activity of the ispA gene is altered to reduce the production of isoprenoid molecules in recombinant cells engineered to produce isoprene or in cells otherwise susceptible to isoprenoid accumulation during fermentation. This decreased ispA gene functional activity enables enhanced synthesis of isoprene in a host microorganism. 1. Recombinant cells capable of producing of isoprene , wherein said cells comprise an ispA gene having decreased functional activity and one or more nucleic acids encoding:(a) an isoprene synthase polypeptide, wherein the isoprene synthase polypeptide is encoded by a heterologous nucleic acid; and(b) one or more mevalonate (MVA) pathway polypeptides,wherein culturing of said recombinant cells in a suitable media provides for the production of said polypeptides and synthesis of isoprene.238-. (canceled)39. A composition comprising the cells of .40. A method of producing isoprene comprising: (a) culturing said recombinant cells of in suitable conditions which provide for production of isoprene; and (b) producing isoprene.4155-. (canceled) This application is a continuation of U.S. patent application Ser. No. 13/725,929, filed Dec. 21, 2012, which claims priority to U.S. Provisional Application No. 61/580,163, filed Dec. 23, 2011 and U.S. Provisional Application No. 61/639,855, filed Apr. 27, 2012, the disclosures of each of which are incorporated by reference herein in their entireties.The Sequence Listing submitted in an ASCII text file, in accordance with 37 C.F.R. §1.821(c) and (e), is incorporated by herein by reference. The text file name is “643842004200.txt”, the date of creation of the text file is Dec. 21, 2012, and the size of the ASCII text file in bytes is 65,536.The present invention relates generally to methods for producing ...

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Номер записи: 69
27-07-2017 дата публикации

Novel Isoprene Synthase and Method of Preparing Isoprene Using Thereof

Номер: US20170211056A1
Автор: Huuskonen Anne, Iimen Marja, JO Jae Hoon, Jung Simon Moon Geun, Koivistoinen Outi, LEE Sang Min, OJA Merja, Ruohonen Laura
Принадлежит:

Provided are a novel isoprene synthase derived from sweet potato and a method of preparing isoprene using the same, and more specifically, a novel isoprene synthase derived from sweet potato, a gene encoding the isoprene synthase, a host cell transformed with the gene, and a method of preparing isoprene using the same. The isoprene synthase of the present invention may have higher isoprene productivity as compared to isoprene synthases known in the related art to thereby be effectively used in isoprene biosynthesis and preparation of an isoprene polymer using the same. 1. An isoprene synthase , which comprises the amino acid of SEQ ID NO: 1; or comprises an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 1.2. The isoprene synthase of claim 1 , wherein the isoprene synthase comprising an amino acid sequence having at least 70% identity to the amino acid of SEQ ID NO: 1 has at least 90% overall identity to positions of SEQ ID NO: 1 at 257-266 (RDRLMESFFW) claim 1 , 287-298 (FKLVTVLDDVYD) claim 1 , 369 (F) claim 1 , 394-396 (SVS) claim 1 , 433-455 (FRLANDLSSSKAEIERGTANSI) claim 1 , and 515-521 (YQYGDAH).3. The isoprene synthase of claim 2 , wherein the isoprene synthase comprising an amino acid sequence having at least 70% identity to the amino acid of SEQ ID NO: 1 has at least one amino acid residue selected from the group consisting of W266 claim 2 , F287 claim 2 , F369 claim 2 , F433 claim 2 , N453 claim 2 , and Y515 of SEQ ID NO: 1.4Ipomoea batatas.. The isoprene synthase of claim 1 , wherein the isoprene synthase is derived from5. (canceled)6. (canceled)7. (canceled)8. (canceled)9. (canceled)10. (canceled)11. A recombinant vector wherein a polynucleotide encoding the isoprene synthase of is operably introduced.12. The recombinant vector of claim 11 , wherein the recombinant vector comprises a promoter for expressing said polynucleotide.13. The recombinant vector of claim 12 , wherein the promoter is selected from the group ...

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Номер записи: 70
27-07-2017 дата публикации

Method for Producing Isoprene Using Recombinant Halophilic Methanotroph

Номер: US20170211100A1
Автор: Cho Kwang Kuk, Kalyuzhnaya Marina, La Yeon Hwa, Lee Ki Sung, Song JaeYang
Принадлежит:

The present invention relates to a recombinant methanotroph having an ability to produce isoprene and a method for producing isoprene using the same, and more particularly to a recombinant methanotroph having an ability to produce isoprene wherein a gene encoding an isoprene synthase having a homology of at least 70% to the amino acid sequence of isoprene synthase is introduced into the recombinant methanotroph, and a method for producing isoprene using the recombinant methanotroph. The use of a recombinant methanotroph according to the present invention enables isoprene to be produced in high yield by using methane gas or methanol which is obtained from waste such as natural gas, biomass, municipal waste or the like as a carbon source. 1. A recombinant methanotroph having an ability to produce isoprene , into which a gene encoding an isoprene synthase having a homology of at least 70% to an amino acid sequence of SEQ ID NO: 1 is introduced.2Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylocystis, Methylomicrobium, Methanomonas, Methylocella,Methylocapsa.. The recombinant methanotroph of claim 1 , wherein the methanotroph is selected from the group consisting of and3Methylomicrobium alcaliphilum.. The recombinant methanotroph of claim 2 , wherein the methanotroph is4. A method for producing isoprene claim 2 , the method comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) culturing the recombinant methanotroph of in the presence of methane or methanol as a carbon source, thereby produce isoprene; and'}(b) recovering the produced isoprene.5. The method of claim 4 , wherein the recovery of isoprene in step (b) is performed by any one method selected from the group consisting of gas chromatography (GC) claim 4 , gas chromatography-mass spectrometry (GC-MS) claim 4 , gas stripping claim 4 , distillation claim 4 , polymer membrane separation claim 4 , adsorption/desorption by pervaporation claim 4 , thermal desorption claim 4 , ...

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Номер записи: 71
04-07-2019 дата публикации

Novel Isoprene Synthase and Method of Preparing Isoprene Using Thereof

Номер: US20190203193A1
Автор: Huuskonen Anne, Ilmen Marja, JO Jae Hoon, Jung Simon MoonGeun, Koivistoinen Outi, LEE Sang Min, OJA Merja, Ruohonen Laura
Принадлежит:

Provided are a novel isoprene synthase derived from sweet potato and a method of preparing isoprene using the same, and more specifically, a novel isoprene synthase derived from sweet potato, a gene encoding the isoprene synthase, a host cell transformed with the gene, and a method of preparing isoprene using the same. The isoprene synthase of the present invention may have higher isoprene productivity as compared to isoprene synthases known in the related art to thereby be effectively used in isoprene biosynthesis and preparation of an isoprene polymer using the same. 1. An isoprene synthase , which comprises the amino acid of SEQ ID NO: 1; or comprises an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 1.2. The isoprene synthase of claim 1 , wherein the isoprene synthase comprising an amino acid sequence having at least 70% identity to the amino acid of SEQ ID NO: 1 has at least 90% overall identity to positions of SEQ ID NO: 1 at 257-266 (RDRLMESFFW) claim 1 , 287-298 (FKLVTVLDDVYD) claim 1 , 369 (F) claim 1 , 394-396 (SVS) claim 1 , 433-455 (FRLANDLSSSKAEIERGTANSI) claim 1 , and 515-521 (YQYGDAH).3. The isoprene synthase of claim 2 , wherein the isoprene synthase comprising an amino acid sequence having at least 70% identity to the amino acid of SEQ ID NO: 1 has at least one amino acid residue selected from W266 claim 2 , F287 claim 2 , F369 claim 2 , F433 claim 2 , N453 claim 2 , and Y515 of SEQ ID NO: 1.4Ipomoea batatas.. The isoprene synthase of claim 1 , wherein the isoprene synthase is derived from This application is a divisional application of U.S. patent application Ser. No. 15/329,343, filed Jan. 26, 2017, which was the United States national phase of International Application No. PCT/KR2015/007851 filed Jul. 28, 2015, which claims priority to Korean Patent Application No. 10-2014-0095972 filed Jul. 28, 2014, the disclosures of which are hereby incorporated in their entirety by reference.The Sequence Listing ...

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Номер записи: 72
04-07-2019 дата публикации

PRODUCTION OF MONOTERPENE BLENDS BY UNICELLULAR PHOTOSYNTHETIC MICROORGANISMS

Номер: US20190203232A1
Автор: Formighieri Cinzia, Melis Anastasios
Принадлежит:

This invention provides compositions and methods for providing high product yield of transgenes expressed in cyanobacteria and microalgae. 1. A method of obtaining a blend of monoterpene hydrocarbons from cyanobacteria , the method comprising:culturing a cyanobacteria strain that has been genetically modified to express a heterologous β-phellandrene synthase under conditions in which the β-phellandrene synthase is expressed, wherein the heterologous β-phellandrene synthase is encoded by a polynucleotide comprising a β-phellandrene synthase nucleic acid sequence that is codon-optimized for expression in cyanobacteria fused to the 3′ end of a leader nucleic acid sequence encoding a cyanobacteria protein that is expressed in cyanobacteria at a level of at least 1% of the total cellular protein; andisolating a blend of monoterpene hydrocarbons comprising at least two monoterpenes selected from the group consisting of β-phellandrene, α-phellandrene, β-myrcene, β-pinene, and δ-2-carene produced in the cyanobacteria that has spontaneously diffused from the cyanobacteria intracellular space into the culture medium.2. The method of claim 1 , wherein the monoterpene blend comprises at least three monoterpenes selected from the group consisting of β-phellandrene claim 1 , α-phellandrene claim 1 , β-myrcene claim 1 , β-pinene claim 1 , and δ-2-carene.3. The method of claim 1 , wherein the β-phellandrene synthase comprises: (i) an arginine-rich region comprising a sequence RR(×8)W claim 1 , wherein (×8) is a sequence of eight amino acids in which each of the positions in the eight-amino acid sequence can be any amino acid; (ii) a C-terminal α-domain active site sequence DDxxD (SEQ ID NO:33) claim 1 , wherein each of the positions designated by “x” can be any amino acid; and (iii) a metal binding sequence (N/D)Dxx(S/T)xxxE (SEQ ID NO:34) claim 1 , wherein each of the positions designated by “x” can be any amino acid.4. The method of claim 1 , wherein the β-phellandrene synthase ...

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Номер записи: 73
05-08-2021 дата публикации

PROCESS OF PRODUCING MONOTERPENES

Номер: US20210238640A1
Автор: KARUPPIAH Vijaykumar, LEFERINK Nicole G. H., Scrutton Nigel S.
Принадлежит:

The present invention relates to a process of producing a monoterpene and/or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and/or derivatives thereof and a recombinant microor-ganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by ex-pression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from 1. A process of producing linalool in a host microorganism comprising the following steps:(a) providing a host microorganism genetically modified to express a bacterial monoterpene synthase (mTS), an acetoacetyl-CoA thiolase (AtoB), a hydroxymethylglutaryl-CoA synthase (HMGS), a hydroxymethylglutaryl-CoA reductase (HMGR), a mevalonate kinase (MK), a phosphomevalonate kinase (PMK), a phosphomevalonate decarboxylase (PMD) and an isopentenyldiphosphate isomerase (idi);(b) converting acetyl CoA into isopentenyl diphosphate (IPP);(c) converting IPP and dimethylallyl diphosphate (DMAPP) into geranyl pyrophosphate (GPP); and(d) contacting GPP with said bacterial mTS to produce linalool,wherein the bacterial mTS comprises linalool synthase, and{'sub': 'org', 'sup': '−1', 'wherein the process results in a linalool yield of at least 100 mg L.'}2E. coli.. The process according to wherein the host microorganism comprises3Streptomyces. The process according to wherein the bacterial mTS comprises a linalool synthase.4Streptomyuces clavuligerus. The process according to wherein the bacterial mTS comprises a linalool synthase.5S. cerevisiae. The process according to wherein the hydroxymethylglutaryl-CoA ...

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Номер записи: 74
13-08-2015 дата публикации

BIOCATALYST FOR CONVERSION OF METHANE AND METHANOL TO ISOPRENE

Номер: US20150225743A1
Автор: DONALDSON Gail K., HOLLANDS Kerry, Picataggio Stephen K.
Принадлежит:

Meythylotrophic cells and in particular methanotrophic bacterial cells are genetically engineered to produce isoprene from methane and/or methanol by expressing a heterologous isoprene synthase, and increasing activity of isopentenyl diphosphate isomerase. In addition, upstream DXP pathway enzymes may have increased activity, enzymes in pathways downstream of IPP and DMAPP may have decreased activity, and methane/methanol assimilation pathway enzymes may have increased activity. 1. Recombinant methylotrophic cells comprising:a) at least one heterologous nucleic acid molecule encoding an isoprene synthase polypeptide; andb) at least one genetic modification which increases isopentenyl diphosphate isomerase activity in the cells as compared with isopentenyl diphosphate isomerase activity in the cells lacking said genetic modification:wherein the cells produce more isoprene when grown in culture conditions comprising at least one of methane and methanol as a carbon source, as compared to the cells without (a) and (b).2. The recombinant cells of wherein the methylotrophic cells are methanotrophic bacterial cells.3. The recombinant cells of wherein the methylotrophic cells are methylotrophic yeasts.4Candida, Hansenula, Pichia, TorulopsisRhodotorula.. The methylotrophic yeasts of selected from the group of genera consisting of claim 3 , and5. The cells of wherein the at least one genetic modification of (b) is accomplished by a process selected from the group consisting of:a) increasing expression of an endogenous polypeptide having isopentenyl diphosphate isomerase activity;b) expressing a heterologous nucleic acid molecule encoding a polypeptide having isopentenyl diphosphate isomerase activity; andc) both (a) and (b).6. The cells of wherein the isoprene synthase polypeptide belongs to the enzyme classification group EC 4.2.3.27.7. The cells of wherein the isopentenyl diphosphate isomerase activity is provided by an isopentenyl diphosphate isomerase polypeptide ...

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Номер записи: 75
13-08-2015 дата публикации

Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene

Номер: US20150225744A1
Автор: Andrew C. Eliot, Caroline M. Peres, Dmitrii V. Vaviline, Zachary Q. Beck
Принадлежит: Goodyear Tire and Rubber Co

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes.

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Номер записи: 76
03-08-2017 дата публикации

Production of Steviol Glycosides in Microorganisms

Номер: US20170218419A1
Автор: Blom Charlotte, Hansen Esben Halkjaer, Hansen Jorgen, HICKS Paula M., Houghton-Larsen Jens, Kishore Ganesh M., Mikkelsen Michael Dalgaard, Motion Michael, Tavares Sabina
Принадлежит:

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements. 1. A recombinant host cell capable of producing steviol , a target steviol glycoside or a target steviol glycoside composition , comprising: 'wherein the polypeptide is capable of transferring a sugar moiety to the C2′ of a glucose in the precursor steviol glycoside;', '(a) a gene encoding a polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of a precursor steviol glycoside;'}and one or more of:(b) a gene encoding a polypeptide capable of glycosylating steviol or the precursor steviol glycoside at its C-13 hydroxyl group; and/or(c) a gene encoding a polypeptide capable of beta 1,3 glycosylation of the C3′ of the 13-O-glucose of the precursor steviol glycoside; and/or(d) a gene encoding a polypeptide capable of glycosylating steviol or the precursor steviol glycoside at its C-19 carboxyl group;wherein at least one of the genes is a recombinant gene.2. The recombinant host cell of claim 1 , wherein:(a) the precursor steviol glycoside is rubusoside, wherein the sugar moiety is glucose, and stevioside is produced upon transfer of the glucose moiety;(b) the precursor steviol glycoside is stevioside, the sugar moiety is glucose, and rebaudioside E is produced upon transfer of the glucose moiety;(c) the precursor steviol glycoside is stevioside, the sugar moiety is glucose, the stevioside is contacted with the polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of the precursor steviol glycoside and a polypeptide ...

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Номер записи: 77
02-07-2020 дата публикации

MODULATION OF CARBON FLUX THROUGH THE MEG AND C3 PATHWAYS FOR THE IMPROVED PRODUCTION OF MONOETHYLENE GLYCOL AND C3 COMPOUNDS

Номер: US20200208160A1
Автор: KOLLING Veronica Maria Rodege Gogola, MAGALHAES Beatriz Leite, Parizzi Lucas Pedersen, Zeidler Ane Fernanda Beraldi
Принадлежит:

The present disclosure provides methods of modulating the flux of carbon through the monoethylene glycol (MEG) biosynthesis pathway and one or more C3 compound biosynthesis pathways by expressing enzymes that are essential for improving C3 compounds and modulating other genetic aspects of MEG and C3 compound biosynthesis. The disclosure is further drawn to modified microbes comprising the disrupted sequences and overexpressed sequences, and compositions thereof. 2. A recombinant microbe capable of coproducing MEG and one or more C3 compounds , wherein the microbe comprises one or more of the following(i) a disruption of one or more exogenous polynucleotide sequences encoding a phosphate acetyltransferase,(ii) a disruption of one or more endogenous polynucleotide sequences encoding an acetate kinase,(iii) a disruption of one or more endogenous polynucleotide sequences encoding a pyruvate oxidase,(iv) a disruption of one or more endogenous polynucleotide sequences encoding an ArcA regulator,(v) a disruption of one or more endogenous polynucleotide sequences encoding a lysine acetyltransferase,(vi) one or more overexpressed endogenous polynucleotide sequences encoding a CobB regulator, and(vii) one or more overexpressed endogenous polynucleotide sequences encoding an acetyl-CoA synthetase;wherein the MEG and/or the one or more C3 compounds are produced at a faster titer, rate or exhibit an increased yield; as compared to a microbe lacking the disruption and/or the overexpression of the endogenous polynucleotides of any one or more of i-vii.3. A recombinant microbe capable of coproducing MEG and one or more C3 compounds , wherein the microbe comprises one or more of the following(i) one or more exogenous polynucleotide sequences encoding a xylose dehydrogenase,(i) one or more exogenous polynucleotide sequences encoding a xylonolactonase,(iii) one or more exogenous polynucleotide sequences encoding a xylonate dehydratase,(iv) one or more exogenous polynucleotide ...

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Номер записи: 78
02-07-2020 дата публикации

PRODUCTION OF BETA-PHELLANDRENE USING GENETICALLY ENGINEERED PHOTOSYNTHETIC MICROORGANISMS

Номер: US20200208178A1
Автор: DAVIES Fiona K., Melis Anastasios, Wintz Hsu-Ching Chen, ZURBRIGGEN Andreas
Принадлежит:

The present invention provides methods and compositions for producing β-phellandrene hydrocarbons from a photosynthetic microorganism such as cyanobacteria.

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Номер записи: 79
20-08-2015 дата публикации

Genetically Enhanced Cyanobacteria for the Production of Isoprene

Номер: US20150232884A1
Автор: DUEHRING Ulf
Принадлежит: ALGENOL BIOFUELS INC.

A cyanobacterium for the production of isoprene having an extrachromosomal plasmid harboring a gene for the production of isoprene. Such a cyanobacterium exhibits a higher isoprene production rate than other conventional strains. 1. A genetically enhanced cyanobacterium capable of producing isoprene , comprising an extrachromosomal plasmid comprising a gene encoding an enzyme that catalyzes the production of isoprene , wherein said gene encoding the enzyme that catalyzes the production of isoprene is under the transcriptional control of a promoter that comprises a ribosomal binding site and a −10 and −35 region from different cyanobacterial promoters.2. The genetically enhanced cyanobacterium of claim 1 , wherein said extrachromosomal plasmid is an endogenous plasmid.3. The genetically enhanced cyanobacterium of claim 1 , wherein said extrachromosomal plasmid is a heterologous plasmid.4. The genetically enhanced cyanobacterium of claim 1 , wherein the promoter is inducible or constitutive.5. The genetically enhanced cyanobacterium of claim 4 , wherein the promoter is selected from the group consisting of P claim 4 , P claim 4 , P claim 4 , and P.6. The genetically enhanced cyanobacterium of claim 1 , wherein the promoter comprises a ribosomal binding site from an isiA-promoter and a −10 and −35 region from a psaA promoter.7. The genetically enhanced cyanobacterium of claim 6 , wherein the promoter sequence comprises SEQ ID NO: 1 claim 6 ,further wherein the 3′-ATG is the start codon of the gene encoding the enzyme that catalyzes the production of isoprene transcriptionally controlled by this promoter.8. The genetically enhanced cyanobacterium of claim 4 , wherein the promoter comprises a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO: 2 claim 4 , further wherein there is 100% sequence identity at nucleotides 4-9 claim 4 , 26-31 claim 4 , and 45-51 of SEQ ID NO: 2.9. The genetically enhanced cyanobacterium of claim 1 , further comprising a ...

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Номер записи: 80
11-08-2016 дата публикации

MICROORGANISMS FOR EFFICIENT PRODUCTION OF MELATONIN AND RELATED COMPOUNDS

Номер: US20160230200A1
Автор: Borodina Irina, Chen Xiao, Forster Jochen, JENSEN Niels Bjerg, Zhu Jiangfeng
Принадлежит:

Recombinant microbial cells and methods for producing 5HTP, melatonin and related compounds using such cells are described. More specifically, the recombinant microbial cell may comprise exogenous genes encoding one or more of an L-tryptophan hydroxylase, a 5-hydroxy-L-tryptophan decarboxylyase, a serotonin acetyltransferase, an acetylserotonin O-methyltransferase; and means for providing tetrahydrobiopterin (THB), and can be further genetically modified to enrich one or more of tryptophan, S-adenosyl-L-methinonine and acetyl coenzyme A. Related sequences and vectors for use in preparing such recombinant microbial cells are also described. 1. A recombinant microbial cell comprising exogenous nucleic acid sequences encoding an L-tryptophan hydroxylase (TPH) (EC 1.14.16.4) , a 5-hydroxy-L-tryptophan decarboxylase (DDC) (EC 4.1.1.28) , a serotonin acetyltransferase (AANAT) (EC 2.3.1.87 or EC 2.3.1.5) , an acetylserotonin O-methyltransferase (ASMT) (EC 2.1.1.4) , and enzymes providing at least one pathway for producing tetrahydrobiopterin (THB) , wherein the recombinant microbial cell(i) further comprises a genetic modification providing for an increase in S-adenosyl-L-methinonine (SAM) production, an increase in acetyl coenzyme A (AcCoA) production, an increase in tryptophan production, or a combination of any thereof; and/or(ii) comprises an exogenous nucleic acid sequence encoding a TPH which comprises SEQ ID NO:177, SEQ ID NO:176, or functionally active variant, homolog or fragment of any thereof.2. The recombinant microbial cell of claim 1 , wherein the genetic modification in (i) comprises one or more exogenous nucleic acid sequences encoding(a) a S-adenosylmethionine synthetase (EC 2.5.1.6),(b) a ethionine resistance protein,(c) a S-adenosylhomocysteine hydrolase (EC 3.3.1.1),(d) a methionine synthase (EC 2.1.1),(e) an AcCoA synthetase (EC 6.2.1.1),(f) an acetylaldehyde dehydrogenase (EC 1.2.1.3), or(g) a combination of any two or more of (a) to (f).3. A ...

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Номер записи: 81
18-07-2019 дата публикации

METHODS, SYNTHETIC HOSTS AND REAGENTS FOR THE BIOSYNTHESIS OF ISOPRENE AND DERIVATIVES

Номер: US20190218577A1
Автор: Cartman Stephen Thomas, FOSTER Alexander Brett, Roberts Ana Teresa De Santos Brito Mendes, Taylor Mark Paul
Принадлежит:

Methods and compositions for synthesizing dienes and derivative thereof, such as isoprene, in are provided. 1Cupriavidus necator. A method for synthesizing isoprene in , said method comprising enzymatically converting isopentenyl-pyrophosphate to dimethylallylpyrophosphate using a polypeptide having isopentenyl diphosphate isomerase enzyme activity.2. The method of wherein the polypeptide having isopentenyl diphosphate isomerase enzyme activity has at least 70% sequence identity to an amino acid sequence set forth in any of SEQ ID NOs: 1 claim 1 , 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 or 6 or a functional fragment thereof.3. The method of wherein the polypeptide having isopentenyl diphosphate isomerase enzyme activity comprises the amino acid sequence set forth in any of SEQ ID NOs: 1 claim 1 , 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 or 6 or a functional fragment thereof.4. The method of wherein the polypeptide having isopentenyl diphosphate isomerase enzyme activity is encoded by a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence set forth in any of SEQ ID NOs: 8 claim 1 , 9 claim 1 , 10 claim 1 , 11 claim 1 , 12 or 13 or a functional fragment thereof.5. The method of wherein the polypeptide having isopentenyl diphosphate isomerase enzyme activity is encoded by a nucleic acid sequence comprising the nucleic acid sequence set forth in SEQ ID NOs: 8 claim 1 , 9 claim 1 , 10 claim 1 , 11 claim 1 , 12 or 13 or a functional fragment thereof.6Cupriavidus necator. A method for synthesizing isoprene in claim 1 , said method comprising enzymatically converting dimethylallylpyrophosphate to isoprene using a polypeptide having isoprene synthase enzyme activity.7. The method of wherein the polypeptide having isoprene synthase enzyme activity has at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO: 7 or a functional fragment thereof.8. The method of wherein the polypeptide having isoprene synthase enzyme ...

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Номер записи: 82
19-08-2021 дата публикации

MICROBIAL PRODUCTION OF ROTUNDONE

Номер: US20210254107A1
Автор: Donald Jason, KUMARAN Ajikumar Parayil, PHILIPPE RYAN N., SANTOS Christine Nicole S., SARRIA STEPHEN
Принадлежит:

The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of α-guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others. 1. A microbial host cell for producing rotundone , the microbial cell expressing a heterologous α-guaiene synthase enzyme (αGTPS) and a heterologous α-guaiene oxidase (αGOX) enzyme.2. The microbial cell of claim 1 , further expressing a farnesyl diphosphate synthase.3. The microbial cell of claim 2 , wherein the αGTPS enzyme comprises an amino acid sequence of any one of SEQ ID NOs: 1 to 21 claim 2 , or variant thereof.4. The microbial cell of claim 3 , wherein α-guaiene synthase enzyme comprises an amino acid sequence having 50% or more sequence identity with any one of SEQ ID NOs: 1 to 21.5. The microbial cell of claim 3 , wherein the αGTPS enzyme comprises an amino acid sequence having 50% or more sequence identity to SEQ ID NO: 8.6. The microbial cell of claim 5 , wherein the αGTPS enzyme comprises one or more amino acid substitutions at positions selected from 72 claim 5 , 273 claim 5 , 290 claim 5 , 368 claim 5 , 371 claim 5 , 374 claim 5 , 377 claim 5 , 381 claim 5 , 382 claim 5 , 399 claim 5 , 406 claim 5 , 419 claim 5 , 433 claim 5 , 442 claim 5 , 443 claim 5 , 454 claim 5 , 512 claim 5 , and 522 relative to SEQ ID NO: 8.7. The microbial cell of claim 6 , wherein the αGTPS enzyme comprises one or more amino acid substitutions selected from T72I claim 6 , M273L claim 6 , R290K claim 6 , F368M claim 6 , I371L claim 6 , S374A claim 6 ...

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Номер записи: 83
25-07-2019 дата публикации

Diterpene production in yarrowia

Номер: US20190226000A1
Автор: Adam G. LAWRENCE, Nicolette Jasmijn Broers, Viktor Marius BOER
Принадлежит: DSM IP ASSETS BV

The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism of the genus Yarrowia in a suitable fermentation medium at a temperature of about 29° C. or higher, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol; and b. recovering the diterpene or glycosylated diterpene.

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Номер записи: 84
23-08-2018 дата публикации

Steviol glycoside transport

Номер: US20180235263A1
Автор: Eric Van Den Berg, Priscilla ZWARTJENS, Viktor Marius BOER
Принадлежит: DSM IP ASSETS BV

A recombinant host capable of producing a steviol glycoside which overexpresses a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto. A recombinant host capable of producing a steviol glycoside which has been modified, preferably in its genome, to result in a deficiency in the production of a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto.

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Номер записи: 85
24-08-2017 дата публикации

Production of Steviol Glycosides in Recombinant Hosts

Номер: US20170240942A1
Автор: Ambri Francesca, Carlsen Simon, Dalgaard Mikkelsen Michael, Dannow Dyekjaer Jane, Douchin Veronique, Halkjær Hansen Esben, Hallwyl Swee Chuang Lim, Houghton-Larsen Jens, Lunde Robertson Helene, Nordmark Andersen Iben, Quiros Asensio Manuel, Rasmussen Nina Nicole, Takos Adam Matthew
Принадлежит:

The invention relates to recombinant microorganisms and methods for producing steviol glycosides and steviol glycoside precursors. 1. A recombinant host comprising one or more of:(a) a gene encoding an ent-kaurene oxidase (KO) polypeptide;(b) a gene encoding a cytochrome P450 reductase (CPR) polypeptide; and/or(c) a gene encoding an ent-kaurenoic acid hydroxylase (KAH) polypeptide;wherein at least one of the genes is a recombinant gene; andwherein the recombinant host is capable of producing a steviol glycoside precursor.2. A recombinant host comprising:(a) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide;(b) a gene encoding an ent-copalyl diphosphate synthase (CDPS) polypeptide;(c) a gene encoding an ent-kaurene synthase (KS) polypeptide(d) a gene encoding an ent-kaurene oxidase (KO) polypeptide;(e) a gene encoding a cytochrome P450 reductase (CPR) polypeptide; and(f) a gene encoding an ent-kaurenoic acid hydroxylase (KAH) polypeptide;wherein at least one of the genes is a recombinant gene; andwherein the recombinant host is capable of producing steviol.3. The recombinant host of or , wherein:(a) the KO polypeptide comprises a KO polypeptide having at least 60% identity to an amino acid sequence set forth in SEQ ID NO:72 or SEQ ID NO:75; at least 65% identity to an amino acid sequence set forth in SEQ ID NO:54; at least 70% identity to an amino acid sequence set forth in SED ID NO: 70, SEQ ID NO:71, or SEQ ID NO:79; at least 40% identity to an amino acid sequence set forth in SEQ ID NO:77; or at least 50% identity to an amino acid sequence set forth in SEQ ID NO:78;(b) the CPR polypeptide comprises a CPR polypeptide having at least 70% identity to an amino acid sequences set forth in SEQ ID NO:69, SEQ ID NO:74, SEQ ID NO:76, or SEQ ID NO:87; at least 80% identity to an amino acid sequence set forth in SEQ ID NO:73; at least 85% identity to an amino acid sequence set forth in SEQ ID NO:22; at least 65% identity to an amino acid sequence set ...

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Номер записи: 86
01-08-2019 дата публикации

METHODS AND MATERIALS FOR THE BIOSYNTHESIS OF DIOL ALCOHOLS AND RELATED COMPOUNDS

Номер: US20190233848A1
Автор: Bawdon Daniel, FOSTER Alexander Brett, KADI Nadia Fatma
Принадлежит:

Methods and materials for the production of diol alcohols, such as 1,2-propanediol (1,2-PD) and derivatives and compounds related thereto. Also provided are products produced in accordance with these methods and materials. 1: A process for biosynthesis of 1 ,2-propanediol (1 ,2-PD) , and/or derivatives thereof and/or compounds related thereto , said process comprising:obtaining an organism capable of producing 1,2-PD, derivatives thereof and/or compounds related thereto;altering the organism; andproducing more 1,2-PD, derivatives thereof and/or compounds related thereto by the altered organism as compared to the unaltered organism.2: The process of wherein the organism is C. necator or an organism with properties similar thereto.3: The process of wherein the organism is altered to express one or more enzymes of glycerol dehydrogenase claim 1 , 1 claim 1 ,2-propanediol oxidoreductase claim 1 , methylglyoxal reductase claim 1 , methylglyoxal synthase and/or dihydroxyacetone kinase.47-. (canceled)8E. coliS. CerevisiaeClostridium acetobutylicumCitrobacter freundii.: The process of wherein the glycerol dehydrogenase and/or 1 claim 3 ,2-propanediol oxidoreductase is from claim 3 , the methylglyoxal reductase is from claim 3 , the methylglyoxal synthase is from or C. necator claim 3 , and/or the dihydroxyacetone kinase is from9: The process of wherein the glycerol dehydrogenase comprises SEQ ID NO:2 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO: 2 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:1 or SEQ ID NO:3 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NOs: 1 or 3 or a functional fragment thereof.1011-. (canceled)12: The process of wherein the 1 claim 3 ,2-propanediol oxidoreductase ...

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Номер записи: 87
08-09-2016 дата публикации

MODIFIED ISOPRENE SYNTHASE

Номер: US20160257944A1
Автор: KASHIMA YASUHIRO, Nakata Kunio, Oku Takashi, Tagami Uno, Tajima Yoshinori
Принадлежит: AJINOMOTO CO., INC.

Modified isoprene synthases that have one or more mutations of a given amino acid residue(s) in any amino acid sequence of 1. A modified isoprene synthase that has one or more mutations of an amino acid residue corresponding to one or more amino acid residues selected from the following:S2, F31, L35, L67, I90, V97, L114, Q123, D124, V125, K127, R128, K130, D131, G134, C137, V144, V187, R202, L224, L247, S248, L249, T257, K259, R264, D265, R266, E269, C286, K292, T298, I300, D301, Y304, D305, E312, F316, E321, V325, I328, C338, L340, C370, E371, C373, E379, S383, K386, I388, A390, Y394, S401, S402, G404, V405, L414, C416, C440, R444, N447, R458, T461, T462, T466, E471, C480, K481, R484, K492, V499, P505, F508, I518, S519, H520, C521, Y523, and G530 in any amino acid sequence of(a) the amino acid sequence of SEQ ID NO:4,(b) an amino acid sequence having one or several amino acid substitutions, deletions, insertions, or additions in the amino acid sequence of SEQ ID NO:4, or(c) an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO:4,and has an isoprene synthetic activity.2. The modified isoprene synthase according to claim 1 , wherein said mutation is selected from the group consisting of S2A claim 1 , F31E claim 1 , F31K claim 1 , L35E claim 1 , L35K claim 1 , L67E claim 1 , 190E claim 1 , V97E claim 1 , L114E claim 1 , Q123C claim 1 , D124E claim 1 , V125E claim 1 , K127D claim 1 , K127E claim 1 , K127N claim 1 , K127R claim 1 , R128K claim 1 , K130G claim 1 , D131C claim 1 , G134P claim 1 , C137E claim 1 , C137I claim 1 , C137L claim 1 , C137M claim 1 , C137S claim 1 , V144E claim 1 , V187E claim 1 , R202D claim 1 , R202N claim 1 , L224E claim 1 , L247D claim 1 , L247E claim 1 , L247Q claim 1 , S248E claim 1 , S248K claim 1 , S248R claim 1 , L249E claim 1 , L249K claim 1 , L249R claim 1 , L249T claim 1 , T257L claim 1 , K259D claim 1 , K259E claim 1 , K259N claim 1 , K259Q claim 1 , K259R claim 1 , R264F claim 1 , R264M claim 1 , ...

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Номер записи: 88
08-08-2019 дата публикации

METHOD FOR PRODUCTION OF ISOPRENE RECOMBINANT MICROORGANISM, GENE CONSTRUCT, VECTOR AND APPLICATION THEREOF

Номер: US20190241913A1
Автор: Ahuja Manmeet, Ambalal Jain Dharmendra, Balkisan Dhoot Shrikant, Isar Jasmine, JOSHI HARSHVARDHAN, Krishna Reddy Chigullarevu Hari, Pankaj Vora Mansi, Prafull Sethia Piyush, Rangaswamy Vidhya, Sanjay Badle Sneh, Suman Varavadekar Jayesh, Suresh Rathod Rachana
Принадлежит:

The present disclosure relates to method for enhanced production of metabolite including but not limiting to isoprene and isoprenoid through chromosomal integration of genes belonging to MEP pathway. The disclosure further relates a host cell for the production of the said metabolite. The method of the present disclosure bypasses the cumbersome method of plasmid application for the production of metabolite. The disclosure also relates to a gene construct comprising MEP genes and auxotrophic markers and a vector comprising the said gene construct. 1. A method for enhanced production of metabolite selected from a group comprising isoprene and isoprenoid , or a combination thereof , said method comprising steps of: transforming host cell with gene construct comprising 2-C-methyl-D-erythritol 4-phosphate (MEP) genes; and integrating the MEP genes into chromosome of the host cell , thereby enhancing production of the metabolite selected from the group comprising isoprene and isoprenoid.2. The method as claimed in claim 1 , wherein the host cell is selected from a group comprising bacteria and fungi.3E. coliE. coliE. coli. The method as claimed in claim 2 , wherein the host cell is selected from a group comprising K-12 claim 2 , K-12 MG1655 and BL21.4. The method as claimed in claim 1 , wherein the MEP genes is selected from a group comprising dxs claim 1 , fni claim 1 , idi claim 1 , dxr claim 1 , ispG claim 1 , ispS claim 1 , ispD claim 1 , and ispF claim 1 , or any combination thereof.5. The method as claimed in claim 4 , the MEP genes are fused by fusion PCR prior to transformation into the host cell claim 4 , wherein the MEP genes are fused in a combination selected from a group comprising—isps and fni;isps, fni and dxs;isps, fni, dxs, dxr andisps, fni, dxs, ispD;6. The method as claimed in claim 5 , wherein the fused MEP genes comprises auxotrophic markers selected from a group comprising thyA claim 5 , metB claim 5 , glnA claim 5 , trpA claim 5 , leuA claim 5 , ...

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Номер записи: 89
30-07-2020 дата публикации

HOST CELLS AND METHODS FOR PRODUCING TRICYCLIC SESQUITERPENES, AVIATION AND MISSILE FUEL PRECURSORS

Номер: US20200239796A1
Автор: Lee Taek Soon
Принадлежит: The Regents of the University of California

The present invention provides for a fuel compositions are provided comprising a hydrogenation product of a tricyclic sesquiterpene (epi-isozizaene, pentalenene, or α-isocomene) and a fuel additive. Methods of making and using the fuel compositions are also disclosed. 1. A genetically modified host cell capable of producing one or more tricyclic sesquiterpenes , said genetically modified host cell comprising one or more tricyclic sesquiterpenes synthase.2. The genetically modified host cell of claim 1 , wherein (a) the tricyclic sesquiterpene is epi-isozizaene claim 1 , and the tricyclic sesquiterpene synthase is epi-isozizaene synthase (EIZS);(b) the tricyclic sesquiterpene is pentalenene, and the tricyclic sesquiterpene synthase is pentalenene synthase (PentS); or (c) the tricyclic sesquiterpene is α-isocomene, and the tricyclic sesquiterpene synthase is α-isocomene synthase (MrTPS2); wherein the EIZS comprises (i) an amino acid sequence having at least 70% identity with SEQ ID NO:1, and (ii) the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; the PentS comprises (i) an amino acid sequence having at least 70% identity with SEQ ID NO:2, and (ii) the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5; and the MrTPS2 comprises (i) an amino acid sequence having at least 70% identity with SEQ ID NO:3, and (ii) the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:5.3. The genetically modified host cell of claim 2 , wherein the tricyclic sesquiterpene is epi-isozizaene claim 2 , and the tricyclic sesquiterpene synthase is epi-isozizaene synthase (EIZS).4. The genetically modified host cell of claim 2 , wherein the tricyclic sesquiterpene is pentalenene claim 2 , and the tricyclic sesquiterpene synthase is pentalenene synthase (PentS).5. The genetically modified host cell of claim 2 , wherein the tricyclic sesquiterpene is α-isocomene claim 2 , and the tricyclic sesquiterpene synthase is α-isocomene synthase (MrTPS2).6. The genetically modified host cell of claim 1 , ...

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Номер записи: 90
15-08-2019 дата публикации

Method for Producing Objective Substance

Номер: US20190249205A1
Автор: Asari Sayaka, FUKUI Keita, KELLY Peter, Mijts Benjamin, Roche Christine, Toyazaki Miku
Принадлежит:

A method for producing an objective substance such as vanillin and vanillic acid is provided. An objective substance is produced from a carbon source or a precursor of the objective substance by using a microorganism having an objective substance-producing ability, which microorganism has been modified so that the activity of an L-cysteine biosynthesis enzyme is increased. 1. A method for producing an objective substance , the method comprising the following step: producing the objective substance by using a microorganism having an ability to produce the objective substance , wherein the microorganism has been modified so that the activity of an L-cysteine biosynthesis enzyme is increased as compared with a non-modified microorganism , and wherein the objective substance is selected from the group consisting of:(X) metabolites the biosynthesis of which requires S-adenosylmethionine, (Y) L-methionine, and (Z) combinations thereof.2. The method according to claim 1 , wherein said producing comprises: cultivating the microorganism in a culture medium containing a carbon source to produce and accumulate the objective substance in the culture medium.3. The method according to claim 1 , wherein said producing comprises: converting a precursor of the objective substance into the objective substance by using the microorganism.4. The method according to claim 3 , wherein said converting comprises: cultivating the microorganism in a culture medium containing the precursor to produce and accumulate the objective substance in the culture medium.5. The method according to claim 3 , wherein said converting comprises: allowing cells of the microorganism to act on the precursor in a reaction mixture to produce and accumulate the objective substance in the reaction mixture.6. The method according to claim 5 , wherein the cells are cells present in a culture broth of the microorganism claim 5 , cells collected from the culture broth claim 5 , cells present in a processed product of ...

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Номер записи: 91
21-10-2021 дата публикации

Method for producing isoprene

Номер: US20210324362A1
Автор: Hui Jeong Jang, Jung Hun KIM, Myeong Seok CHA, Seon Won Kim
Принадлежит: Condair Group AG, Industry Academic Cooperation Foundation of GNU

A method for producing isoprene includes culturing E. coli, which has isoprene productivity and in which a gene encoding a recA protein is attenuated or deleted, in a medium containing a carbon source. Therefore, a great amount of isoprene may be produced within a short period of time, and thereby considerably decreasing isoprene production unit costs.

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Номер записи: 92
04-12-2014 дата публикации

Novel sesquiterpene synthase gene and protein

Номер: US20140356926A1
Автор: Chappell Joseph, Greenhagen Bryan
Принадлежит:

The invention relates to sesquiterpene synthases and methods for their production and use. Particularly, the invention provides nucleic acids comprising the nucleotide sequence of citrus valencene synthase (CVS) which codes for at least one CVS. The invention further provides nucleic acids comprising the nucleotide sequence coding for amino acid residues forming the tier 1 and tier 2 domains of CVS. The invention also provides for methods of making and using the nucleic acids and amino acids of the current invention. 1. A method of producing valencene , comprising reacting a valencene synthase polypeptide with a farnesyl pyrophosphate substrate under conditions in which the valencene synthase polypeptide reacts with the farnesyl pyrophosphate substrate to catalyze the production of valencene , wherein the valencene synthase polypeptide comprises the sequence of amino acid residues set forth in SEQ ID NO:4.2. The method of claim 1 , wherein the reaction of the polypeptide with farnesyl pyrophosphate is performed at pH between about 6.0 and 7.0.3. The method of claim 1 , wherein the valencene synthase polypeptide is expressed in a host cell that comprises heterologous nucleic acid encoding the valencene synthase polypeptide.4. The method of claim 3 , wherein the host cell is selected from among a yeast cell claim 3 , a bacterial cell claim 3 , an insect cell and a plant cell.5. The method of claim 3 , further comprising isolating the host cell.6. The method of claim 3 , wherein the nucleic acid molecule is a vector.7. The method of claim 6 , wherein the vector is a viral vector claim 6 , bacteriophage or a plasmid.8. The method of claim 1 , further comprising reacting the valencene by regiospecific hydroxylation and then oxidation to produce nootkatone.9. The method of claim 8 , further comprising isolating the nootkatone.10. The method of claim 8 , wherein the regiospecific hydroxylation and oxidation are catalyzed by a single multifunctional hydroxylase.11. The ...

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Номер записи: 93
29-09-2016 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCING ISOPRENE FREE OF C5 HYDROCARBONS UNDER DECOUPLING CONDITIONS AND/OR SAFE OPERATING RANGES

Номер: US20160281112A1
Автор: CALABRIA Anthony R., Cervin Marguerite A., Chotani Gopal K., La Duca Richard, McAuliffe Joseph C., Miller Michael C., SABO Timothy A., SANFORD Karl J., WEBSTER Erin L., Whited Gregory M.
Принадлежит:

The invention features methods for producing isoprene from cultured cells wherein the cells in the stationary phase. The invention also provides compositions that include these cultured cells and/or increased amount of isoprene. The invention also provides for systems that include a nonflammable concentration of isoprene in the gas phase. Additionally, the invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity. 122-. (canceled)23. A method of producing isoprene , the method comprising(a) culturing recombinant cells comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide under suitable culture conditions for the production of isoprene, wherein the amount of isoprene produced during stationary phase is greater than or about 2-fold more than the amount of isoprene produced during growth phase for the same length of time; and(b) producing isoprene.24. The method of claim 23 , wherein the cells are cultured under limited glucose conditions.25. The method of claim 23 , further comprising (c) recovering the isoprene.2631-. (canceled)32. The method of claim 25 , wherein the isoprene is recovered from an off-gas portion of the cell culture.33. The method of claim 23 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.34. The method of claim 23 , wherein the cells are bacterial cells.35Escherichia coli, Pantoae citrea, Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus clausii, Bacillus halodurans, Bacillus megaterium, Bacillus coagulans, Bacillus circulans, Bacillus lautus, Bacillus thuringiensis, Streptomyces lividans, Streptomyces coelicolor, Streptomyces griseus, PseudomonasPseudomonas alcaligenes. The method of claim 34 , wherein the bacterial cells are selected from the group consisting of sp. claim 34 , and cells.36. The method of claim ...

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Номер записи: 94
29-09-2016 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCING ISOPRENE

Номер: US20160281113A1
Автор: Cervin Marguerite A., Chotani Gopal K., Feher Frank J., La Duca Richard, McAuliffe Joseph C., Miasnikov Andrei, PERES Caroline M., Puhala Aaron S., SANFORD Karl J., Valle Fernando, Whited Gregory M.
Принадлежит:

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. 120-. (canceled)21. Recombinant cells capable of producing isoprene , the cells comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide , wherein the cells (i) produce greater than about 400 nmole/g/hr of isoprene , (ii) convert more than about 0.002 molar percent of the carbon that the cells consume from a cell culture medium into isoprene , or (iii) have an average volumetric productivity of isoprene greater than about 0.1 mg/L/hr.22. The cells of claim 21 , wherein the heterologous nucleic acid encoding an isoprene synthase polypeptide is operably linked to a promoter.23. The cells of claim 21 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.24. The cells of claim 23 , wherein the plant isoprene synthase polypeptide is a poplar isoprene synthase polypeptide or a kudzu isoprene synthase polypeptide.25. The cells of claim 21 , wherein the heterologous nucleic acid encoding an isoprene synthase polypeptide is in a vector or is integrated into a chromosome of the cells.26. The cells of claim 21 , further comprising nucleic acids encoding: (a) an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide or (b) at least one of a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide and/or one or more mevalonate (MVA) pathway polypeptides.27. The cells of claim 26 , wherein at least one of the nucleic acids encoding one or more MVA pathway polypeptides of (b) is a heterologous nucleic acid or a copy of an endogenous nucleic acid.28. The cells of claim 26 , wherein the cells comprise polypeptides of the entire MVA pathway.29. The cells of claim 26 , wherein at least one of the nucleic acids encoding a polypeptide of (a) and (b) is in a vector or is integrated into a chromosome of the cells.30. The cells of claim 26 , wherein at least one of the nucleic acids ...

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Номер записи: 95
27-09-2018 дата публикации

OLFACTORY LIGANDS

Номер: US20180271089A1
Автор: ALLEMANN Rudolf Konrad, Birkett Michael Alexander, MILLER David James, Pickett John Anthony
Принадлежит:

The invention provides analogues of (S)-germacrene D analogue which have improved insect repellent properties compared to (S)-germacrene D analogue or which have insect attractant properties. 2. The compound according to wherein claim 1 , independently or in any combination:{'sup': '1', 'Ris H, methyl or ethyl;'}{'sup': '2', 'Ris H, methyl or ethyl;'}{'sup': '3', 'Ris methyl or ethyl;'}{'sup': '4', 'Ris H, methyl or ethyl;'}{'sup': '5', 'Ris H, methyl or ethyl.'}3. The compound according to wherein:{'sup': 1', '2', '4, 'each of R, Rand Ris H;'}{'sup': '3', 'Ris methyl, ethyl, n-propyl, iso-propyl or cyclopropyl; and'}{'sup': '5', 'Ris H, methyl, ethyl, n-propyl, iso-propyl or cyclopropyl.'}4. The compound according to wherein Ris H.5. The compound according to which is (S)-15-methylgermacrene D.6. The compound according to wherein each of R claim 1 , Rand Ris independently methyl claim 1 , ethyl claim 1 , n-propyl claim 1 , iso-propyl or cyclopropyl and each of Rand Ris H;7. The compound according to wherein Ris H.8. The compound according to which is ((S)-14 claim 7 ,15-dimethylgermacrene D.10. The process according to wherein the germacrene D synthase is a recombinant (S)-germacrene D synthase polypeptide.11. The process according to wherein the recombinant GDS comprises a tag sequence at the N- or C-terminus claim 10 , in particular a polyhistidine tag.12. The process according to wherein the GDS comprises a C-terminal polyhistidine tag claim 11 , for example a hexahistidine tag.13Solidago canadensis. The process according to wherein the GDS is native germacrene D synthase from SEQ ID NO: 1.14Solidago canadensis. The process according to wherein the GDS is native germacrene D synthase from SEQ ID NO: 1 claim 9 , which has one or more of the modifications:tyrosine residue at position 406 replaced by phenylalanine, leucine, isoleucine, valine or alanine;tryptophan residue at position 275 replaced by phenylalanine, leucine, isoleucine, valine or alanine, but ...

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Номер записи: 96
08-10-2015 дата публикации

RECOMBINANT CELL, AND METHOD FOR PRODUCING ISOPRENE

Номер: US20150284742A1
Автор: Fischer Rainer, FURUTANI Masahiro, Iwasa Koichiro, Jennewein Stefan, Uenishi Akihiro
Принадлежит:

To provide a series of techniques capable of producing isoprene from syngas or the like. 120-. (canceled)21. A recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell having an isopentenyl diphosphate synthesis ability by a non-mevalonate pathway , wherein the nucleic acid is expressed in the host cell , and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of carbon monoxide , carbon dioxide , formic acid , and methanol.22. The recombinant cell according to claim 21 , having carbon monoxide dehydrogenase.23ClostridiumMoorella. The recombinant cell according to claim 21 , wherein the host cell is a bacterium or a bacterium.24. The recombinant cell according to claim 21 , wherein a nucleic acid encoding a group of enzymes acting in a mevalonate pathway is further introduced so that an isopentenyl diphosphate synthesis ability by a mevalonate pathway is further imparted.25. The recombinant cell according to claim 24 , wherein the mevalonate pathway is that of yeast claim 24 , prokaryote or actinomycete.26. The recombinant cell according to claim 21 , wherein a nucleic acid encoding at least one enzyme acting in a non-mevalonate pathway is further introduced claim 21 , and the nucleic acid is expressed in the host cell.27. The recombinant cell according to claim 26 , wherein the non-mevalonate pathway is that of other organism than the host cell.28. The recombinant cell according to claim 21 , wherein the isoprene synthase is derived from plant.29. The recombinant cell according to claim 21 , wherein the nucleic acid encoding isoprene synthase encodes the following (a) claim 21 , (b) or (c):(a) a protein consisting of an amino acid sequence represented by SEQ ID NO: 2,(b) a protein consisting of an amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 2, and having isoprene ...

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Номер записи: 97
08-10-2015 дата публикации

METHODS FOR THE ENZYMATIC PRODUCTION OF ISOPRENE FROM ISOPRENOL

Номер: US20150284743A1
Автор: Anissimova Maria, Marliere Philippe
Принадлежит:

Described are methods for the enzymatic production of isoprene which allow to produce isoprene from isoprenol and microorganisms which have been genetically modified so as to be able to produce isoprene from isoprenol. 1. A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into isoprenyl monophosphate and then enzymatically converting isoprenyl monophosphate into isoprene.2. The method of wherein the enzymatic conversion of isoprenol into isoprenyl monophosphate is achieved by the use of a hydroxyethylthiazole kinase (EC 2.7.1.50).3. The method of wherein isoprenyl monophosphate is directly enzymatically converted into isoprene (Pathway A).4. The method of wherein the enzymatic conversion of isoprenyl monophosphate into isoprene is achieved by the use of a terpene synthase.5. The method of wherein the terpene synthase is selected from an isoprene synthase (EC 4.2.3.27) claim 4 , a monoterpene synthase claim 4 , an alpha-farnesene synthases (EC 4.2.3.46) claim 4 , an beta-farnesene synthase (EC 4.2.3.47 claim 4 ,) a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15) or a pinene synthase (EC 4.2.3.14).6. The method of wherein isoprenyl monophosphate is converted enzymatically into prenyl monophosphate by an isomerisation reaction and prenyl monophosphate is then converted into isoprene by a dephosphorylation reaction (Pathway B).7. The method of wherein the enzymatic conversion of isoprenyl monophosphate into prenyl monophosphate is achieved by the use of an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2) and wherein the enzymatic conversion of prenyl monophosphate into isoprene is achieved by the use of a terpene synthase.8. A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into prenol by an isomerisation reaction claim 6 , and then enzymatically converting prenol into prenyl monophosphate by a phosphorylation reaction and then enzymatically converting prenyl monophosphate into ...

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Номер записи: 98
28-09-2017 дата публикации

GENETICALLY MODIFIED (R)-LACTIC ACID PRODUCING THERMOPHILIC BACTERIA

Номер: US20170275656A1
Автор: MACHIELSEN Marinus Petrus, VAN HARTSKAMP Mariska, VAN KRANENBURG Richard
Принадлежит: Purac Biochem BV

The invention relates to a genetically engineered thermophilic bacterial cell that is facultative anaerobic comprising: a) inactivation or deletion of the endogenous (S)-lactate dehydrogenase gene; b) introduction of a (R)-lactate dehydrogenase gene; c) inactivation or deletion of the endogenous pyruvate formate lyase A and/or B gene. 1. A genetically engineered thermophilic bacterial cell that is facultative anaerobic comprising:a) inactivation or deletion of the endogenous (S)-lactate dehydrogenase gene;b) introduction of a (R)-lactate dehydrogenase gene;c) inactivation or deletion of the endogenous pyruvate formate lyase A and/or B gene.2. The cell according to wherein in addition the endogenous methylglyoxal synthase gene mgsA is inactivated or deleted.3Lactobacillus delbrueckii. The cell according to wherein the (R)-lactate dehydrogenase is the hdhD gene from encoding the amino acid sequence of SEQ ID NO:38 or an amino acid sequence having at least 90% identity.4Lactobacillus delbrueckii. The cell according to wherein the (R)-lactate dehydrogenase is the ldhA gene from encoding the amino acid sequence of SEQ ID NO:36 or an amino acid sequence having at least 90% identity.5. The cell according to wherein the hdhD gene encodes the amino acid sequence of SEQ ID NO:38.6. The cell according to wherein the ldhA gene encodes the amino acid sequence of SEQ ID NO:36.7. The cell according to wherein in addition the endogenous phosphotransacetylase gene (pta) is inactivated or deleted.8. The cell according to which is a sporulation deficient derivative due to inactivation or deletion of an endogenous sporulation gene.9. The cell according to wherein the sporulation gene is sigF.10. The cell according to wherein the pyruvate formate lyase A and/or B gene is inactivated by inactivation or deletion of the endogenous pyruvate formate lyase/alcohol dehydrogenase locus pflBA-adhE.11. The cell according to which produces (R)-lactic acid with an enantiomeric purity of at least 98 ...

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Номер записи: 99
27-09-2018 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCING ISOPRENE

Номер: US20180273982A1
Автор: Cervin Marguerite A., Chotani Gopal K., Feher Frank J., La Duca Richard, McAuliffe Joseph C., Miasnikov Andrei, PERES Caroline M., Puhala Aaron S., SANFORD Karl J., Valle Fernando, Whited Gregory M.
Принадлежит:

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. 120-. (canceled)221. The method of claim , wherein the cells further comprise an endogenous nucleic acid encoding an isoprene synthase polypeptide.231. The method of claim , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.244. The method of claim , wherein the plant isoprene synthase polypeptide is a poplar isoprene synthase polypeptide or a kudzu isoprene synthase polypeptide.251. The method of claim , wherein the cells further comprise nucleic acids encoding: (a) an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide or (b) a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide.261. The method of claim , wherein at least one of the nucleic acids encoding one or more MVA pathway polypeptides is a heterologous nucleic acid or a copy of an endogenous nucleic acid.271. The method of claim , wherein the cells comprise polypeptides of the entire MVA pathway.281. The method of claim , wherein the cells are fungal , bacterial , plant , or algal cells.298. The method of claim , wherein the cells are fungal cells.309AspergillusTrichoderma. The method of claim , wherein the cells are , yeast , or , cells.3110SaccharomycesSchizosaccharomycesPichiaYarrowiaCandida. The method of claim , wherein the yeast cells are sp. , sp. , sp. , sp. , or sp. cells329A. oryzae, A. niger, S. cerevisiae, S. pombe, T. reesei, H. insolens, H. lanuginose, H. grisea, C. lucknowense, A. sojae, A. japonicus, A. nidulans, A. aculeatus, A. awamori, F. roseum, F. graminum, F. cerealis, F. oxysporuim, F. venenatum, N. crassa, M. miehei, T. viride, F. oxysporumF. solan. The method cells of claim , where the cells are selected from the group consisting of , and cells.338. The method of claim , wherein the cells are bacterial cells.3413. The method of claim , wherein the cells are gram-positive bacterial cells or ...

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Номер записи: 100