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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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Применить Всего найдено 1099. Отображено 100.
17-01-2013 дата публикации

Synthesis of Prazole Compounds

Номер: US20130017580A1
Автор: Benjamin Mijts, Derek Smith, Jovana Nazor, Jun Zhu, Michael D. Clay, Michael Vogel, Shiwei Song, Steven J. Collier, Xiyun Zhang, Yong Koy Bong
Принадлежит: Codexis Inc

The present disclosure relates to non-naturally occurring monooxygenase polypeptides useful for preparing prazole compounds, polynucleotides encoding the polypeptides, and methods of using the polypeptides.

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Номер записи: 1
17-01-2013 дата публикации

Nucleic acid construct comprising pyripyropene biosynthetic gene cluster and marker gene

Номер: US20130017581A1
Автор: Hiroyuki Anzai, Kentaro Yamamoto, Koichiro Murashima, Kouji Yanai, Naomi Sumida, Sato Aihara
Принадлежит: Individual

There is provided a nucleic acid construct comprising a pyripyropene biosynthetic gene cluster and a marker gene. The nucleic acid construct according to the present invention provides an inexpensive and highly productive method for producing pyripyropene.

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Номер записи: 2
01-08-2013 дата публикации

Modulation of nitric oxide signaling to normalize tumor vasculature

Номер: US20130195926A1
Автор: Dai Fukumura, Rakesh K. Jain, Satoshi Kashiwagi
Принадлежит: General Hospital Corp

The instant invention provides methods for treating a solid tumor in a subject comprising modulating nitric oxide production in the tumor to normalize tumor vasculature and administering an anti-tumor therapy to the subject. The invention further provides methods of treating a solid tumor in a subject comprising selectively increasing cyclic guanosine monophosphate (cGMP) or cGMP dependent protein kinase G production in the tumor vasculature to an amount effective to normalize tumor vasculature and administering an anti-tumor therapy to the subject.

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Номер записи: 3
03-01-2019 дата публикации

BIOFILMS, COMPONENTS AND METHODS OF USE TO REDUCE BIOFOULING AND CONTAMINATION

Номер: US20190000090A1
Автор: Kumar Manish, Wood Thammajun L., Wood Thomas K.
Принадлежит:

Biofilms are provided which are capable of regulating their own thickness, reducing contamination and preventing biofouling. Constructs are introduced into bacteria that comprise nucleic acid molecules encoding an autoinducer synthase polypeptide, a transcriptional regulator and a biofilm dispersal protein. Nucleic acid molecules may also be introduced which encode a nitric oxide synthase, an epoxide hydrolase, or both. Biofilms of the bacteria may be used to reduce biofouling and contamination of a surface. 1. A method of producing a living self-controlled biofilm of engineered bacteria cells on a surface , said method comprising: i. a nucleic acid molecule encoding an autoinducer synthase polypeptide;', 'ii. a nucleic acid molecule encoding a transcriptional regulator, capable of being activated by said autoinducer synthase polypeptide; and', 'iii. a nucleic acid molecule encoding a biofilm dispersal protein to produce said at least one engineered bacteria cell comprising said construct; and, 'a) producing at least one engineered bacteria cell by introducing into said at least one bacterial cell a quorum sensing nucleic acid construct, comprising,'}b) producing a self-controlled biofilm of said at least one engineered bacteria-cells, wherein said quorum sensing nucleic acid construct in response to activation of said transcriptional regulatory and production of said biofilm dispersal protein reduces the thickness of said self-controlled biofilm compared to biofilm not comprising said quorum sensing nucleic acid construct.24.-. (canceled)5. The method of claim 1 , further comprising introducing into said at least one bacterial cell a nucleic acid molecule encoding nitric oxide synthase.6. The method of claim 1 , further comprising introducing into said bacterial cell a nucleic acid molecule encoding epoxide hydrolase.78.-. (canceled)9. The method of claim 1 , wherein said thickness of said biofilm is at least six fold less than said biofilm not comprising said ...

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Номер записи: 4
07-01-2021 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20210002618A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 136 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 136 , selected from the group consisting of 75 , 79 , 82 , 99 , 110 , 166 , 172 , 208 , 216 , 273 , 324 , 364 , 395 , 412 , 491 , 503 , and 504.3. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 136.5. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: ...

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Номер записи: 5
01-01-2015 дата публикации

COMPOSITIONS AND METHODS OF ALTERING CHOLESTEROL LEVELS

Номер: US20150005372A1
Автор: de Fougerolles Antonin, Ellsworth Jeff Lynn, HOGE Stephen G.
Принадлежит:

The present invention relates to compositions, methods and kits using polynucleotides, primary transcripts and mmRNA molecules. 1. A composition comprising;(a) a synthetic polynucleotide encoding PCSK9 negative LDLR, and(b) a synthetic polynucleotide encoding CYP7A1, in an acceptable diluent or carrier.2. The composition of claim 1 , further comprising;(c) a statin selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, simvastatin, and combinations thereof.3. The composition of claim 1 , wherein the synthetic polynucleotide comprises:(a) a first region of linked nucleosides, said first region encoding at least one cholesterol regulating polypeptide; '(i) a sequence of linked nucleosides selected from the group consisting of the native 5′ untranslated region (UTR), SEQ ID NO: 1 and functional variants thereof;', '(b) a first flanking region located at the 5′ terminus of said first region comprising;'} (i′) a sequence of linked nucleosides selected from the group consisting of the native 3′ UTR, any of the forgoing comprising one or more microRNA or microRNA binding site or microRNA seeds and functional variants or combinations thereof; and', '(ii′) a 3′ tailing sequence of linked nucleosides;', 'wherein the first region of linked nucleosides comprises at least a first modified nucleoside., '(c) a second flanking region located at the 3′ terminus of said first region comprising;'}4. The composition of wherein the second flanking region encodes at least one miR binding site.5. The composition of claim 4 , wherein said miR binding site is selected from the group consisting of miR-122a and miR-422a.6. The composition of any of wherein the synthetic polynucleotide comprises at least one chemical modification.7. A method of treating a disease or disorder in a subject in need thereof comprising administering to said subject the composition of any of .8. The method of further comprising measuring ...

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Номер записи: 6
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: 7
12-01-2017 дата публикации

METABOLIC TRANSISTOR IN BACTERIA

Номер: US20170009241A1
Автор: Bennett George N., San Ka-Yiu, Wu Hui
Принадлежит:

The disclosure relates to a metabolic transistor in microbes such as bacteria and yeast where a competitive pathway is introduced to compete with a product pathway for available carbon so as to control the carbon flux in the microbe. 1) A method of increasing the production of a product in a microbe , said method comprising:a) providing a microbe having a product pathway producing a product, wherein a competitive pathway competes with said product pathway for available carbon, said competitive pathway requiring a cofactor that is not rate limiting and not used in said product pathway;b) adding one or more diverting gene(s) under the control of a promoter to said microbe to directly or indirectly divert said cofactor away from said competitive pathway; and,c) allowing expression of said diverting gene(s) and reducing levels of said cofactor, and thus reducing levels of said competitive pathway;d) thereby increasing said product pathway and said product.4) The method of claim 1 , wherein said promoter is a constitutive promoter.5) The method of claim 1 , wherein said promoter is an inducible promoter.6) A method of controlling aerobic respiration in a microbe in the presence of O claim 1 , said method comprising:a) adding a diverting gene(s) to a microbe to divert substrates away from ubiquinone or thiamine or heme production, wherein said diverting gene(s) is under the control of a promoter; and,{'sub': '2', 'b) inducing said promoter, thereby allowing expression of said diverting gene(s), thus reducing ubiquinone levels and reducing aerobic respiration in the presence of 0.'}7) The method of claim 6 , wherein said diverting gene(s) includes the lycopene synthesis pathway.8) The method of claim 6 , wherein said diverting gene(s) include crtE claim 6 , crtB and crtI.9) The method of claim 6 , wherein said diverting gene encodes geranyl diphosphate:4-hydroxybenzoate geranyltransferase.10) The method of claim 6 , wherein said diverting gene is lePGT-1.11) The method of ...

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Номер записи: 8
03-02-2022 дата публикации

PROGRAMMED MICROORGANISMS TO ATTENUATE A DISEASE

Номер: US20220033867A1
Автор: KUMAR Arumbuliyur Sathish, MURALI Panchapagesa Muthuswamy, Raghavan Shriram
Принадлежит:

The present disclosure discloses a recombinant microbe producing podophyllotoxin, or its derivatives, comprising genes encoding phenyl alanine ammonia-lyase (PAL), cinnamate-4-hydroxylate (C4H), 4-coumaroyl CoA-ligase (4CL), hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HCT), p-coumaroyl quinate 3′-hydroxylase (C3H), caffeoyl CoA O-methyltransferase (CCoAOMT), bifunctional pineresionl-lariciresinol reductase (DIRPLR), secoisolariciresinol dehydrogenase (SDH), cytochrome P450 oxidoreductase CYP719, O-methyltransferase (OMT), cytochrome P450 oxidoreductase CYP71, and 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD). Also disclosed herein is a method for producing podophyllotoxin or its derivatives. Moreover, a method of treating cancer is also disclosed. 1. A recombinant microbe producing podophyllotoxin , or its derivatives , comprising genes encoding phenyl alanine ammonia-lyase (PAL) , cinnamate-4-hydroxylate (C4H) , 4-coumaroyl CoA-ligase (4CL) , hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HCT) , p-coumaroyl quinate 3′-hydroxylase (C3H) , caffeoyl CoA O-methyltransferase (CCoAOMT) , bifunctional pineresionl-lariciresinol reductase (DIRPLR) , secoisolariciresinol dehydrogenase (SDH) , cytochrome P450 oxidoreductase CYP719 , O-methyltransferase (OMT) , cytochrome P450 oxidoreductase CYP71 , and 2-oxoglutarate/Fe(II)-dependent dioxygenase (2-ODD).2. The recombinant microbe as claimed in claim 1 , wherein the recombinant microbe further comprises gene encoding cytochrome P450 oxidoreductase CYP82D.3. The recombinant microbe as claimed in claim 2 , wherein the recombinant microbe further comprises gene encoding UDP glucosyl transferase.4. The recombinant microbe as claimed in claim 3 , wherein the recombinant microbe further comprises gene encoding 2-Deoxy-d-ribose-5-phosphate aldolase.5. The recombinant microbe as claimed in any one of the - claim 3 , wherein two or more genes are fused to encode fusion proteins.6. The recombinant microbe ...

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Номер записи: 9
19-01-2017 дата публикации

TETRACYCLINE RESISTANT EUKARYOTIC CELLS EXPRESSING AN NADP-REQUIRING OXIDOREDUCTASE

Номер: US20170015979A1
Автор: Cardineau Guy A., Echaurri Sergio Ángel García
Принадлежит:

TETX, a tetracycline degrading enzyme, is provided as a selection marker for eukaryotic cells. Polynucleotide molecules containing tetx and DNA constructs comprising a tetx under the control of a eukaryotic promoter are provided. Additionally, DNA constructs that contain a gene of interest and a tetx, each under the control of an appropriate promoter are provided. Further, methods of expressing tetx in a eukaryotic cell are provided, the method comprising introducing a vector comprising tetx into eukaryotic cells, culturing the eukaryotic cells in the presence of tetracycline, and identifying and isolating the cell that expresses tetx. Furthermore, a method is provided for expressing a gene of interest encoding a protein of interest in a eukaryotic cell by culturing a cell containing a vector comprising a gene of interest and a tetx, each under the control of an appropriate promoter. 1. A DNA construct comprising a first promoter operably linked to a tetx , wherein tetx is a polynucleotide comprising:(a) the sequence set forth in any one of SEQ ID NOs: 1-62;(b) a sequence having at least 70% sequence identity to the sequence set forth in any one of SEQ ID NOs: 1-62,wherein the first promoter drives transcription of the tetx when the DNA construct is in a eukaryotic cell.2. The DNA construct of claim 1 , wherein the first promoter is selected from cmv claim 1 , ef1a claim 1 , sv40 claim 1 , pgk1 claim 1 , ubc claim 1 , beta actin claim 1 , beta 2 tubulin claim 1 , hsp70a/rbcs2 claim 1 , introns of rbcs2 claim 1 , cag claim 1 , uas claim 1 , ac5 claim 1 , polyhedrin claim 1 , camkIIa claim 1 , gal1 claim 1 , gal10 claim 1 , tef1 claim 1 , gds claim 1 , adh1 claim 1 , camv35S claim 1 , ubi claim 1 , h1 claim 1 , ponA or u6 promoter.3. The DNA construct of claim 1 , wherein the first promoter is selected from beta 2 tubulin or hsp70a/rbcs2 promoter.4. A eukaryotic expression vector comprising the DNA construct of .5. A eukaryotic cell comprising the DNA construct of .6. ...

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Номер записи: 10
17-01-2019 дата публикации

INCREASED TRIACYLGLYCEROL PRODUCTION IN MICROALGAE

Номер: US20190017082A1
Автор: DOLCH Lina, MARECHAL Eric
Принадлежит:

The application generally relates to bioproduction of molecules of interest in micro- organisms, more particularly in microalgae. In particular, the application relates to methods for increasing triacylglycerol production in micro-organisms, in particular in microalgae, using recombinant micro-organisms which have been genetically engineered to produce or overproduce nitric oxide (NO) and uses thereof. 1. A method for increasing the production of a molecule of interest in a micro-organism , said method comprising:culturing a recombinant micro-organism which has been genetically engineered to produce or overproduce nitric oxide (NO) under conditions suitable for the production or overproduction of NO by said micro-organism, so as to enhance the production of said molecule of interest and recovering said molecule of interest.2. The method according to claim 1 , wherein the molecules of interest are molecules of the lipid metabolic pathway or biomolecules derived from said molecules and wherein said lipid metabolic pathway is comprised between acetyl-CoA and lipids.3. The method according to claim 1 , wherein said micro-organism is a microalga.4. The method according to claim 1 , wherein said micro-organism has been transformed with a recombinant nucleic acid encoding a protein involved in an NO production pathway or wherein expression of an endogenous gene encoding a protein involved in an NO production pathway is modified.5. The method according to claim 1 , wherein said micro-organism has been transformed with a recombinant nucleic acid encoding a NOA protein.6Phaeodactylum tricornutum. The method according to claim 4 , wherein said protein is the NOA (PtNOA) protein or a homolog or functional variant thereof.7. The method according to claim 4 , wherein said protein has at least 80% sequence identity with SEQ ID NO:1.8Nannochloropsis gaditana. The method according to claim 4 , wherein said protein is the NOA (NgNOA) protein or a homolog or functional variant thereof ...

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Номер записи: 11
28-01-2021 дата публикации

Microbial production of steviol glycosides

Номер: US20210022373A1
Автор: Ajikumar Parayil Kumaran, Jason Donald, Krishna PATEL, Liwei Li, Ryan LIM, Ryan PHILIPPE, Swati Gupta
Принадлежит: Manus Bio Inc

The invention provides methods for making steviol glycosides, including RebM and glycosylation products that are minor products in stevia leaves, and provides enzymes, encoding polynucleotides, and host cells for use in these methods. The invention provides engineered enzymes and engineered host cells for producing steviol glycosylation products, such as RebM, at high purity and/or yield. The invention further provides methods of making products containing steviol glycosides, such as RebM, including food products, beverages, oral care products, sweeteners, and flavoring products.

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Номер записи: 12
10-02-2022 дата публикации

METHOD FOR THE BIOSYNTHESIS OF DIOSMIN AND/OR HESPERIDIN IN A MICROORGANISM

Номер: US20220042061A1
Автор: JOUBERT LAETITIA, LE JEUNE ANDRÉ, PAUTHENIER CYRILLE, ROUSSEL CÉLIA, SCORNEC HÉLÈNE
Принадлежит:

The present invention relates to a recombinant microorganism which is modified to be capable of producing diosmin and hesperidin and to the use thereof for producing diosmin and/or hesperidin. 132-. (canceled)33. A recombinant microorganism comprising:a heterologous nucleic acid sequence coding for a flavanone 7-O-beta-D-glucosyltransferase (UGT) which is capable of adding a glucose in position 7 of hesperetin and/or diosmetin; anda heterologous nucleic acid sequence coding for a 6″-O-rhamnosyltransferase (RhaT) which is capable of transferring a rhamnose into position 6 of the glucose of hesperetin-7-O-glucoside and/or diosmetin-7-O-glucoside; anda heterologous nucleic acid sequence coding for a UDP-glucose 4,6-dehydratase/UDP-4-keto-6-deoxy-D-glucose 3,5-epimerase/UDP-4-keto-L-rhamnose-reductase (RHM) which is capable of producing UDP-rhamnose.34. The microorganism as claimed in claim 33 , in which the flavanone 7-O-beta-D-glucosyltransferase is selected from the group consisting of SEQ ID NOs: 113 claim 33 , 115 claim 33 , 91 claim 33 , 93 claim 33 , 95 claim 33 , 97 claim 33 , 99 and 101 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having flavanone 7-O-beta-D-glucosyltransferase activity.35. The microorganism as claimed in claim 33 , in which the 6″-O-rhamnosyltransferase is selected from the group consisting of SEQ ID NOs: 103 and 105 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having 6″-O-rhamnosyltransferase activity.36. The microorganism as claimed in claim 33 , in which the UDP-glucose 4 claim 33 ,6-dehydratase/UDP-4-keto-6-deoxy-D-glucose 3 claim 33 ,5-epimerase/UDP-4-keto-L-rhamnose-reductase is selected from the group consisting of SEQ ID NOs: 107 claim 33 , 109 and 111 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having UDP-glucose 4 claim 33 ,6-dehydratase/UDP-4- ...

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Номер записи: 13
24-01-2019 дата публикации

Herbicide-detoxifying enzymes and uses thereof

Номер: US20190024109A1
Автор: Brian Thompson
Принадлежит: Nufarm Ltd

Herbicide-detoxifying enzymes, compositions containing one or more of the enzymes, and plant seeds treated with the enzymes are provided. The enzymes can be used in methods for detoxifying auxin herbicides or degrading auxin plant regulators, including in methods for decontaminating surface of an apparatus used in agriculture or pesticide manufacturing, methods for decontaminating water, soil, soilless media, or sludge, and methods for protecting a plant from an auxin herbicide, improving a plant's tolerance to an auxin herbicide, or removing an auxin herbicide from the surface of a plant.

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Номер записи: 14
28-01-2021 дата публикации

RNA PREPARATIONS COMPRISING PURIFIED MODIFIED RNA FOR REPROGRAMMING CELLS

Номер: US20210024895A1
Автор: Dahl Gary, Jendrisak Jerome, Kariko Katalin, Meis Judith, Person Anthony, Weissman Drew
Принадлежит:

The present invention provides compositions and methods for reprogramming somatic cells using purified RNA preparations comprising single-strand mRNA encoding an iPS cell induction factor. The purified RNA preparations are preferably substantially free of RNA contaminant molecules that: i) would activate an immune response in the somatic cells, ii) would decrease expression of the single-stranded mRNA in the somatic cells, and/or iii) active RNA sensors in the somatic cells. In certain embodiments, the purified RNA preparations are substantially free of partial mRNAs, double-stranded RNAs, un-capped RNA molecules, and/or single-stranded run-on mRNAs. 118-. (canceled)19. A method for reducing the immunogenicity for mammalian cells of a preparation of in vitro-synthesized RNA molecules comprising:purifying a preparation of in vitro-synthesized RNA molecules obtained by a process comprising in vitro transcription (IVT),{'sup': 1', '5', '5', '2, 'wherein said in vitro-synthesized RNA molecules encode at least one recombinant protein and comprise at least one modified nucleoside selected from ψ, mψ, mU, moU, and sU in place of U,'}wherein said purifying uses a purification process that removes RNA contaminant molecules comprises dsRNA molecules that are toxic to mammalian cells by inducing an innate immune response, and wherein said purifying generates a purified RNA preparation that exhibits reduced immunogenicity,wherein said reduced immunogenicity is detectable using an in vitro MDDC immunogenicity assay by measuring secretion of less IFN-α or TNF-α cytokine secreted by human or murine monocyte-derived dendritic cells (MDDCs) transfected with said purified RNA preparation than is secreted from MDDCs transfected with said preparation of in vitro-synthesized RNA molecules that have not been subjected to said purifying.20. The method of claim 19 , wherein said process comprises replacing at least a portion of the UTP in the IVT reaction mixture by the corresponding ...

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Номер записи: 15
04-02-2016 дата публикации

Cytochrome P450S and Uses Thereof

Номер: US20160032256A1
Автор: Chappell Joseph, Ralston Lyle F.
Принадлежит: University of Kentucky Research Foundation

The invention features isolated cytochrome P450 polypeptides and nucleic acid molecules, as well as expression vectors and transgenic plants containing these molecules. In addition, the invention features uses of such molecules in methods of increasing the level of resistance against a disease caused by a plant pathogen in a transgenic plant, in methods for producing altered compounds, for example, hydroxylated compounds, and in methods of producing isoprenoid compounds. 1. A method for producing in a recombinant host cell an isoprenoid compound not endogenously produced by a non recombinant host cell , comprising: the isoprenoid synthase catalyzes production of an isoprenoid compound;', 'wherein the isoprenoid synthase is a diterpene synthase; and', 'the cytochrome P450 polypeptide(s) catalyzes dual hydroxylation, oxidation, demethylation or methylation of the isoprenoid compound; and, 'a) providing a recombinant host cell that comprises heterologous nucleic acid encoding an isoprenoid synthase, and heterologous nucleic acid encoding one or more protein(s) comprising a cytochrome P450 polypeptide, whereinb) culturing the recombinant host cell under conditions suitable for expressing the isoprenoid synthase and the cytochrome P450 polypeptide(s) under conditions for producing the isoprenoid compound wherein the synthase and the cytochrome P450 polypeptide(s) catalyze formation of the isoprenoid compound in the host cell.2. The method of claim 1 , wherein at least one cytochrome P450 polypeptide has oxidase activity.3. The method of claim 1 , wherein at least one cytochrome P450 polypeptide has dual hydroxylase activity.4. The method of claim 1 , wherein the host cell is a yeast cell claim 1 , a bacterial cell claim 1 , an insect cell or a plant cell.5. The method of claim 1 , wherein the host cell is a yeast cell.6Saccharomyces cerevisiae.. The method of claim 5 , wherein the yeast is7. The method of claim 1 , wherein at least one of the cytochrome P450 polypeptides ...

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Номер записи: 16
01-02-2018 дата публикации

A METHOD FOR UTILIZING ENGINEERED DENDRITIC CELLS TO INDUCE GUT-HOMING REGULATORY T CELLS AND TREAT GUT INFLAMMATION

Номер: US20180030553A1
Автор: Baylink David J., Lau K.-H. William, Tang Xiaolei, Walter Michael
Принадлежит:

Gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme. A method for treating one or more than one inflammation-related condition or disease, the method comprising administering gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme. 1. A gene-modified dendritic cell suitable for treating one or more than one inflammation-related condition or disease; the gene-modified dendritic cell comprising a 1-alpha-hydroxylase gene that produces functional 1-alpha-hydroxylase and a retinaldehyde dehydrogenase 2 gene that produces functional retinaldehyde dehydrogenase 2.2. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is administered to a patient.3. The gene-modified dendritic cell of claim 2 , wherein after administration to the patient the gene-modified dendritic cell migrates into and actively produce 1 claim 2 ,25(OH)2D and retinoid acid in peripheral lymphoid organs.4. A pharmaceutical suitable for treating one or more than one inflammation-related condition or disease claim 2 , the pharmaceutical comprising:a gene-modified dendritic cell comprising a 1-alpha-hydroxylase gene that produces functional 1-alpha-hydroxylase and a retinaldehyde dehydrogenase 2 gene that produces functional retinaldehyde dehydrogenase 2.5. A method for treating one or more than one inflammation-related condition or disease claim 2 , the method comprises:identifying a ...

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Номер записи: 17
17-02-2022 дата публикации

Engineering Bacteria for Ferulic Acid Production, Preparation Method and Use Thereof

Номер: US20220049235A1
Автор: Haili Liu, Huajun Lv, Jie Shao, Ying Zhang, Yong Wang
Принадлежит: Center for Excellence in Molecular Plant Sciences of CAS

The disclosure provides an engineering bacterium for ferulic acid production, a preparation method of the bacterium and use thereof. The invention provides an engineering bacterium that can efficiently produce ferulic compounds by expressing a series of heterologous enzymes in a host cell through gene recombination technology. The expression system constructed by the invention has low metabolic background, strong heterologous expression ability and low cost. The system can synthesize the end product through relatively simple steps, and provide a new way for the industrial production of ferulic acid, intermediates or derivatives thereof.

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Номер записи: 18
17-02-2022 дата публикации

Plants with improved digestibility and marker haplotypes

Номер: US20220049265A1
Автор: Carolina LOPEZ-DURAN, Milena OUZUNOVA, Monika KLOIBER-MAITZ, Nina Meyer, Therese BOLDUAN
Принадлежит: KWS SAAT SE and Co KGaA

The present invention relates to plants, such as maize, Sorghum or sugar cane, having improved digestibility, in particular improved stover digestibility. The present invention relates to a QTL allele associated with improved digestibility and specific marker alleles associated with the QTL allele. The present invention further relates to such plants, wherein the F35H gene is mutated or wherein F35H expression is altered. The invention also relates to methods for identifying plants having improved digestibility and methods for obtaining such plants.

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Номер записи: 19
31-01-2019 дата публикации

FUNCTIONAL EXPRESSION OF MONOOXYGENASES AND METHODS OF USE

Номер: US20190032028A1
Автор: Clark Elizabeth Jane, Greenfield Derek Lorin, Helman Noah Charles, Jones Stephanie Rhianon, Zhu Baolong
Принадлежит:

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

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Номер записи: 20
31-01-2019 дата публикации

CREATION OF CHRYSANTHEMUM WITH BLUE FLOWER COLOR

Номер: US20190032066A1
Автор: Aida Ryutaro, HONGO Satoshi, Noda Naonobu, Sato Sanae, Tanaka Yoshikazu
Принадлежит:

Provided are transformed plants having blue flower color, their self-fertilized progenies or cross-fertilized progenies thereof, a vegetative propagated plants thereof, and a part, a tissue or a cell of the plant body. Anthocyanin 3′,5′-O-glucosyltransferase gene (CtA3′5′GT) derived from and flavonoid 3′,5′-hydroxylase gene derived from (CamF3′5′H) are coexpressed in petals. 1: An expression cassette comprising: (1-a) a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 1;', '(1-b) a polynucleotide that hybridizes with a polynucleotide comprising the nucleotide sequence complementary to the nucleotide sequence listed as SEQ ID NO: 1 under stringent conditions, the polynucleotide encoding a protein with activity of transferring sugars to the and 5′-hydroxyl groups of anthocyanins;', '(1-c) a polynucleotide encoding a protein comprising the amino acid sequence listed as SEQ ID NO: 2;', '(1-d) a polynucleotide comprising an amino acid sequence which is the amino acid sequence listed as SEQ ID NO: 2 with a deletion, substitution, insertion and/or addition of one or more amino acids, and encoding a protein with activity of transferring sugars to the 3′- and 5′-hydroxyl groups of anthocyanins; and', '(1-e) a polynucleotide having an amino acid sequence with at least 90% identity with the amino acid sequence listed as SEQ ID NO: 2, and encoding a protein with activity of transferring sugars to the 3′- and 5′-hydroxyl groups of anthocyanins, and, 'a first polynucleotide selected from the group consisting of the following (1-a) to (1-e) (2-a) a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 3;', '(2-b) a polynucleotide that hybridizes with a polynucleotide comprising the nucleotide sequence complementary to the nucleotide sequence listed as SEQ ID NO: 3 under stringent conditions, the polynucleotide encoding a protein with activity of hydroxylating the 3′- and 5′-positions of flavonoids;', '(2-c) a polynucleotide encoding a protein ...

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Номер записи: 21
30-01-2020 дата публикации

Kaurenoic acid hydroxylases

Номер: US20200032225A1
Автор: Johannes Gustaaf Ernst VAN LEEUWEN, Priscilla ZWARTJENS, Viktor Marius BOER
Принадлежит: DSM IP ASSETS BV

The present invention relates a polypeptide having kaurenoic acid 13-hydroxylase activity, which 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 or SEQ ID NO: 3, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 136, 248, 336 or 403, said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

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Номер записи: 22
08-02-2018 дата публикации

COMPOSITIONS AND METHODS OF ALTERING CHOLESTEROL LEVELS

Номер: US20180036369A1
Автор: de Fougerolles Antonin, Ellsworth Jeff Lynn, HOGE Stephen G.
Принадлежит: Moderna TX, Inc.

The present invention relates to compositions, methods and kits using polynucleotides, primary transcripts and mmRNA molecules. 1. A composition comprising a synthetic polynucleotide encoding PCSK9 negative a low density lipoprotein receptor (LDLR) having one or more amino acids substituted such that the LDLR mutant is deficient in proprotein convertase subtilisin/kexin type 9 (PCSK9) binding , in an acceptable diluent or carrier.2. (canceled)46-. (canceled)7. A method of treating a disease or disorder in a subject in need thereof comprising administering to said subject the composition of .816-. (canceled)17. A method of modulating cholesterol levels in plasma of a subject comprising contacting said subject with the composition of .18. (canceled)19. A polynucleotide comprising an mRNA encoding a LDLR mutant having one or more amino acids substituted such that the LDLR mutant is deficient in PCSK9 binding claim 1 , wherein the one or more amino acids correspond to amino acid 316 of SEQ ID NO: 19 claim 1 , amino acid 317 of SEQ ID NO:19 claim 1 , amino acid 331 of SEQ ID NO:19 claim 1 , amino acid 336 of SEQ ID NO:19 claim 1 , amino acid 339 of SEQ ID NO:19 claim 1 , or any combination thereof.20. The polynucleotide of claim 19 , wherein the LDLR mutant has two or more amino acids substituted such that the LDLR mutant is deficient in PCSK9 binding claim 19 , wherein the two or more substituted amino acids correspond to any two or more of amino acid 316 of SEQ ID NO:19 claim 19 , amino acid 317 of SEQ ID NO:19 claim 19 , amino acid 331 of SEQ ID NO:19 claim 19 , amino acid 336 of SEQ ID NO:19 claim 19 , and amino acid 339 of SEQ ID NO:19.21. The polynucleotide of claim 19 , wherein the substituted amino acids comprise one or more of N316A claim 19 , E317A claim 19 , D331A claim 19 , Y336A claim 19 , L339D claim 19 , D331E claim 19 , or any combination thereof.22. The polynucleotide of claim 19 , wherein the half-life of the LDLR mutant encoded by the polynucleotide is ...

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Номер записи: 23
09-02-2017 дата публикации

SYNTHETIC METHANOTROPHIC AND METHYLOTROPHIC MICROORGANISMS

Номер: US20170037438A1
Автор: CLARKE Elizabeth, GREENFIELD Derek, HELMAN Noah
Принадлежит: INDUSTRIAL MICORBES, INC.

Provided herein are non-naturally occurring microbial organisms comprising a methane-oxidizing metabolic pathway. The invention additionally comprises non-naturally occurring microbial organisms comprising pathways for the production of chemicals. The invention additionally provides methods for using said organisms for the production of chemicals. 1. A synthetic microorganism , wherein said synthetic microorganism comprises a natural methanol-consuming microorganism and one or more genetic modifications that improve the production of a chemical.2. (canceled)3. A synthetic microorganism comprising a natural non-methanol-consuming microorganism and one or more genetic modifications that allow said synthetic microorganism to oxidize methanol.4Escherichia coli, Bacillus subtilis, Pseudomonas putida, Saccharomyces cerevisiae, Corynebacterium glutamicum Klebsiella oxytoca, Anaerobiospirillum succiniciproducens, Actinobacillus succinogenes, Mannheimia succiniciproducens, Rhizobium etli, Gluconobacter oxydans, Zymomonas mobilis, Lactococcus lactis, Lactobacillus plantarum, Streptomyces coelicolor, Clostridium acetobutylicum, Pseudomonas fluorescens, Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces marxianus, Aspergillus terreusAspergillus niger.. The synthetic microorganism of claim 3 , wherein the natural non-methanol-consuming microorganism is selected from the group consisting of claim 3 , and5Corynebacterium glutamicumEscherichia coli.. The synthetic microorganism of claim 4 , wherein the natural non-methanol-consuming microorganism is not or6. The synthetic microorganism of claim 3 , further comprising exogenous polynucleotides claim 3 , wherein said exogenous polynucleotides encode enzymes selected from the group consisting of methanol dehydrogenase (EC 1.1.1.224 or 1.1.99.37 or 1.1.2.7) claim 3 , alcohol dehydrogenase (EC 1.1.1.1) claim 3 , 3-hexulose-6-phosphate synthase (EC 4.1.2.43) and 6-phospho-3-hexuloisomerase (EC 5.3.1.27).7. The synthetic ...

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Номер записи: 24
24-02-2022 дата публикации

Biological-based catalyst to delay plant development processes

Номер: US20220053761A1
Автор: George Pierce, Sidney Crow, JR.
Принадлежит: Crowpierce Technologies LLC

Disclosed are purified proteins, cell lysates, cell-free extracts, and protein extracts comprising 1-hexene monooxygenase (1-HMO) and methods of their use for delaying plant development.

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Номер записи: 25
08-02-2018 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20180037872A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO:136 , and one or more amino acid substitutions at one or more positions in SEQ ID NO: 136 , selected from the group consisting of 37 , 277 , 278 , 280 , 281 , 326 , 432 , 433 , 435 , and 490.3. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO:136.4. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring further is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% enantiomeric excess.5. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions selected from the group consisting of a glycine at position 143 claim 1 , glycine at ...

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Номер записи: 26
08-02-2018 дата публикации

PROTECTING GROUP CHEMISTRY FOR CLEAN, REDUCTANT-FREE DYEING

Номер: US20180037917A1
Автор: BHAKTA Shyam Pravin, CERVANTES Bernardo, DUEBER John Eugene, FONG, HSU Tammy Melody, III Arthur Muir, Johnson, Jr. Terry Don, LATIMER Luke Nathaniel, PRATHURI Ramya Lakshmi, RUSS Zachary Nicholas
Принадлежит: The Regents of the University of California

The present disclosure relates to the biosynthesis of indigoid dye precursors and their conversion to indigoid dyes. Specifically, the present disclosure relates to methods of using polypeptides to produce indigoid dye precursors from indole feed compounds, and the use of the indigoid dye precursors to produce indigoid dyes. 2. The method of claim 1 , wherein the host cell is a bacterial cell.3E. coliCorynebacterium glutamicum.. The method of claim 2 , wherein the host cell is selected from the group consisting of and4. The method of claim 1 , wherein the host cell is a eukaryotic cell.5. The method of claim 4 , wherein the host cell is a yeast.6Saccharomyces cerevisiae, Pichia pastoris, Kluyveromyces lactis, Kluyveromyces marxianus,Schizosaccharomyces pombe.. The method of claim 4 , wherein the host cell is selected from the group consisting of and7. The method of claim 1 , wherein the polypeptide with oxygenase activity comprises SEQ ID NO: 1 claim 1 , SEQ ID NO: 2 claim 1 , SEQ ID NO: 21 claim 1 , SEQ ID NO: 22 claim 1 , SEQ ID NO: 23 claim 1 , or SEQ ID NO: 24 claim 1 , or the amino acid sequence of any homologs thereof.8. The method of claim 1 , wherein the polypeptide with glycosyltransferase activity comprises SEQ ID NO: 3 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , SEQ ID NO: 7 claim 1 , SEQ ID NO: 8 claim 1 , SEQ ID NO: 15 claim 1 , SEQ ID NO: 16 claim 1 , SEQ ID NO: 17 claim 1 , SEQ ID NO: 18 claim 1 , SEQ ID NO: 19 claim 1 , or SEQ ID NO: 20 claim 1 , or the amino acid sequence of any homologs thereof.9. The method of claim 1 , wherein the polypeptide with sulfotransferase activity comprises SEQ ID NO: 9 claim 1 , SEQ ID NO: 10 claim 1 , SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, or SEQ ID NO: 14, or the amino acid sequence of any homologs thereof.10. The method of claim 7 , wherein the homolog of a polypeptide with oxygenase activity comprises an amino acid sequence at least 90% claim 7 , at least 95% claim 7 , or at ...

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Номер записи: 27
07-02-2019 дата публикации

Inhibiting UVB-Irradiation Damage By Targeting Nitric Oxide Synthases (cNOS)

Номер: US20190038723A1
Автор: Tong Lingying, Wu Shiyong
Принадлежит: OHIO UNIVERSITY

Compositions and methods for inhibition UVB-irradiation damage by targeting constitutive nitric oxide synthases (cNOS), including endothelial NOS (eNOS) and neuronal NOS (nNOS) are described. 1. A method of treating a skin cell of a subject , comprising:administering a constitutive nitric oxide synthases (cNOS) inhibitor composition to the subject in an amount sufficient to inhibit DNA damage in the skin cells caused by exposure to UVB irradiation, and/or to protect the skin cells from UVB-induced transformation;wherein the cNOS inhibitor composition comprises an endothelial NOS (eNOS) and/or a neuronal NOS (nNOS).2. The method of claim 1 , wherein the cNOS inhibitor composition comprises at least one of: L-N-nitro-arginine methyl ester (L-NAME) and NG-monomethyl-L-arginine (L-NMMA).3. The method of claim 1 , wherein the composition is in the form of a topical composition.4. The method of claim 3 , wherein the topical composition is in the form of a spray claim 3 , mist claim 3 , aerosol claim 3 , lotion claim 3 , cream claim 3 , solution claim 3 , oil claim 3 , gel claim 3 , ointment claim 3 , paste claim 3 , emulsion claim 3 , suspension injectable and/or transdermal application.5. The method of claim 1 , wherein the skin of the subject is normal and healthy claim 1 , and wherein the composition is applied to the skin for a time and in an amount sufficient to cause a reduction in UV damage to skin cells.6. A composition comprising: an effective amount of a constitutive nitric oxide synthase (cNOS) inhibitor sufficient to inhibit DNA damage in the skin cells caused by exposure to UVB irradiation claim 1 , and/or to protect the skin cells from UVB-induced transformation;wherein the cNOS inhibitor comprises an endothelial NOS (eNOS) and/or a neuronal (nNOS); and,one or more acceptable excipients, diluents and/or carriers and/or encapsulated.7. The composition of claim 6 , wherein the cNOS inhibitor comprises comprises at least one of: L-N-nitro-arginine methyl ester ...

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Номер записи: 28
15-02-2018 дата публикации

AMPHIPATHIC PEPTIDE

Номер: US20180043031A1
Автор: Davison Ashley, McCarthy Helen, Robson Tracy, Zholobenko Aleksey
Принадлежит:

The present invention is directed to an amphipathic peptide and methods of using the amphipathic peptide for delivering small molecule agents to a cell. Ideally, the amphipathic cell penetrating peptide comprises less than approximately 50 amino acid residues with at least 6 arginine residues, at least 12 Alanine Residues, at least 6 leucine residues, optionally at least one cysteine residue, and at least two but no greater than three glutamic acids wherein the arginine residues are evenly distributed along the length of the peptide; and the peptide has a defined ratio of arginine to negatively charged amino acid residues and a defined ratio of hydrophilic amino acid residues to hydrophobic amino acid residues. The present invention is also directed to a nanoparticle and cell delivery system comprising the amphipathic cell penetrating peptide of the invention. The peptide, nanoparticle or cell delivery system of the invention may be used in therapy. For example, the peptide may be used as a therapeutic agent delivery system, in which the therapeutic agent may include nucleic acids or other small molecules. 1. An amphipathic cell penetrating peptide of less than approximately 50 amino acid residues comprising at least 6 arginine residues (R) , at least 12 alanine residues (A) , at least 6 leucine residues (L) , optionally at least one cysteine residue (C) , and at least two but no greater than three glutamic acids (E) whereinthe arginine (R) residues are evenly distributed along the length of the peptide;the ratio of arginine (R) to negatively charged glutamic acid (E) residues is from at least 6:2 to 9:2; andthe ratio of hydrophilic amino acid residues to hydrophobic amino acid residues at pH 7 is at least 30:67 to 40:60; orwherein the peptide comprises or consists of WEARLARALARALARELARALARALRACEA (SEQ ID No. 4).2. An amphipathic cell penetrating peptide of less than approximately 50 amino acid residues comprising at least 6 arginine residues (R) , at least 12 ...

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Номер записи: 29
14-02-2019 дата публикации

PRODUCTION OF STEVIOL GLYCOSIDES IN RECOMBINANT HOSTS

Номер: US20190048356A1
Автор: Douchin Veronique, Takos Adam Matthew
Принадлежит:

The invention relates to recombinant microorganisms and methods for producing steviol glycosides and steviol glycoside precursors. 1. A recombinant host cell capable of producing one or more steviol glycosides or a steviol glycoside composition in a cell culture , comprising a recombinant gene encoding damage resistance protein 1 (DAP1) polypeptide having at least 60% sequence identity to the amino acid sequence set forth in SEQ ID NO:2;wherein expression of the recombinant gene encoding DAP1 polypeptide results in increased production of the one or more steviol glycosides or the steviol glycoside composition in the cell culture.2. The recombinant host cell of claim 1 , further comprising:(a) a gene encoding a polypeptide capable of synthesizing geranylgeranyl pyrophosphate (GGPP) from farnesyl diphosphate (FPP) and isopentenyl diphosphate (IPP);(b) a gene encoding a polypeptide capable of synthesizing ent-copalyl diphosphate from GGPP;(c) a gene encoding a polypeptide capable of synthesizing ent-kaurene from ent-copalyl pyrophosphate;(d) a gene encoding a polypeptide capable of glycosylating steviol or a steviol glycoside at its C-13 hydroxyl group;(e) a gene encoding a polypeptide capable of beta 1,3 glycosylation of the C3′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of a steviol glycoside;(f) a gene encoding a polypeptide capable of glycosylating steviol or a steviol glycoside at its C-19 carboxyl group; and/or(g) 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 steviol glycoside;and 'wherein the polypeptide comprises a polypeptide having at least 70% sequence identity to one of the amino acid sequences set forth in SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:117, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:74, or SEQ ID NO:76; and', '(h) a gene encoding a polypeptide capable of synthesizing ent-kaurenoic acid, ent- ...

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Номер записи: 30
13-02-2020 дата публикации

CULTURE MODIFIED TO CONVERT METHANE OR METHANOL TO 3-HYDROXYPROPRIONATE

Номер: US20200048639A1
Автор: CLARKE Elizabeth Jane, Greenfield Derek Lorin, Helman Noah Charles, Jones Stephanie Rhianon, Zhu Baolong
Принадлежит:

Provided are engineered organisms which can convert methane or methanol to 3-hydroxypropionate. 1. A synthetic culture comprising one or more microorganisms comprising one or more modifications that improve the production of a product from a substrate , wherein the substrate comprises methane and/or methanol.2. The synthetic culture according to claim 1 , wherein the substrate comprises methane.3. The synthetic culture according to claim 2 , wherein the product comprises 3-hydroxyproprionate.4. The synthetic culture according to claim 1 , wherein the product comprises 3-hydroxyproprionate.5. The synthetic culture according to claim 1 , wherein the product comprises a substance derived from acetyl-CoA and/or malonyl-CoA.6Escherichia coli.. The synthetic culture according to claim 1 , wherein at least one of the one or more microorganisms comprises7. The synthetic culture according to claim 1 , wherein the one or more microorganisms comprises a first at least one microorganism and a second at least one microorganism claim 1 , wherein the first at least one microorganism produces methanol from methane and the second at least one microorganism produces 3-hydroxypropionate from methanol.8. The synthetic culture according to claim 1 , wherein the one or more modifications comprise exogenous polynucleotides or deletion of one or more genes.9. The synthetic culture according to claim 8 , wherein the exogenous polynucleotides encode polypeptides selected from one or more polypeptides comprising methane monooxygenase (EC 1.14.13.25) claim 8 , malonyl-CoA reductase (EC 1.2.1.75) claim 8 , acetyl-CoA carboxylase (EC 6.4.1.2) claim 8 , methanol dehydrogenase (EC 1.1.1.244 or EC 1.1.2.7) claim 8 , 3-hexulose-6-phosphate synthase (EC 4.1.2.43) claim 8 , and/or 6-phospho-3-hexuloisomerase (EC 5.3.1.27).10Bacillus methanolicus, Bacillus stearothermophilusCorynebacterium glutamicum.. The synthetic culture according to claim 9 , wherein the methanol dehydrogenase comprises a methanol ...

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Номер записи: 31
10-03-2022 дата публикации

Methods for production of oxygenated terpenes

Номер: US20220073955A1
Автор: Ajikumar Parayil Kumaran, Anthony Qualley, Chin Giaw Lim, Christopher Pirie, Geoff MARSHALL-HILL, Liwei Li, Martin Preininger, Souvik Ghosh
Принадлежит: Givaudan SA, Manus Bio Inc

The present disclosure relates to methods for producing oxygenated terpenoids, and preparation of compositions and formulations thereof. Polynucleotides, derivative enzymes, and host cells for use in such methods are also provided.

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Номер записи: 32
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: 33
05-03-2015 дата публикации

ENZYMATIC ALKENE CLEAVAGE

Номер: US20150064756A1
Автор: Kroutil Wolfgang, Rajagopalan Aashrita, Schober Markus
Принадлежит: UNIVERSITAT GRAZ

The invention relates to an enzyme that comprises or includes a sequence according to SEQ. ID No. 1 or SEQ. ID No. 2, to a method for the production thereof, and to the use thereof as a catalyst in the oxidative cleavage of vinyl aromatics. 1. An enzyme that has or comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.2. Use of an enzyme according to claim 1 , or an enzyme having an amino acid sequence showing at least 80% identity to SEQ ID NO: 1 or 2 claim 1 , as a catalyst for cleaving vinyl aromatics with oxygen.3. The use according to claim 2 , wherein the enzyme has an amino acid sequence showing at least 90% claim 2 , preferably at least 95% claim 2 , more preferably at least 99% claim 2 , identity to SEQ ID NO: 1 or 2.4. The use according to claim 3 , wherein the enzyme has an amino acid sequence showing at least 95% identity to SEQ ID NO: 1 or 2.5. The use according to claim 4 , wherein the enzyme has an amino acid sequence showing at least 99% identity to SEQ ID NO: 1 or 2.6. The use according to claim 2 , wherein cleavage is performed in the presence of Mn3+ ions.7Trametes hirsuta. A method of preparing an enzyme according to claim 1 , wherein a culture of is cultured and the enzyme is recovered from a cell-free extract of said culture.8Trametes hirsuta. The method according to claim 7 , wherein a culture of G FCC 047 is cultured.9. The method according to claim 7 , wherein recovering from the cell-free extract is performed by a combination of hydrophobic interaction chromatography and anion exchange chromatography.10. The method according to claim 9 , wherein hydrophobic interaction chromatography claim 9 , anion exchange chromatography claim 9 , and again hydrophobic interaction chromatography are performed claim 9 , in this order claim 9 , to recover the enzyme.11. A method of recombinantly producing an enzyme according to claim 1 , wherein a nucleic acid encoding the enzyme claim 1 , having a nucleotide sequence according to SEQ ID NO: 3 ...

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Номер записи: 34
27-02-2020 дата публикации

Magnetically immobilized metabolic enzymes and cofactor systems

Номер: US20200061597A1
Автор: Matthew Stephen Chun, Rani Talal BROOKS, Stephane Cedric Corgie
Принадлежит: Zymtronix Catalytic Systems Inc

The present invention provides compositions and methods for producing magnetic bionanocatalysts (BNCs) comprising metabolically self-sufficient systems of enzymes that include P450 monooxygenases or other metabolic enzymes and cofactor regeneration enzymes.

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Номер записи: 35
09-03-2017 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20170067036A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO:2 , and one or more amino acid substitutions at one or more positions in SEQ ID NO: 2 selected from the group consisting of 277 , 281 , 326 , 433 , or 435.3. The non-naturally occurring polynucleotide of claim 1 , wherein said encoded polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 2 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO:2.4. The non-naturally occurring polynucleotide of claim 1 , wherein said encoded polypeptide further is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% enantiomeric excess.5. The non-naturally occurring polynucleotide of claim 1 , wherein said encoded polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 claim 1 , wherein said polypeptide further comprises one or more substitutions selected from the group consisting of a glycine at position 143 claim 1 , glycine at position 278 claim 1 , arginine at position ...

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Номер записи: 36
12-03-2015 дата публикации

NOVEL CAMPANULA FLAVONOID 3',5'-HYDROXYLASE GENE AND ITS USE

Номер: US20150074855A1
Автор: Katsumoto Yukihisa, Tanaka Yoshikazu
Принадлежит:

There is provided a novel flavonoid 3′,5′-hydroxylase gene, and a plasmid comprising the gene under the control of the cauliflower mosaic virus 35S promoter. 1. A polynucleotide selected from the group consisting of the following (a) to (d):(a) a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 1;(b) a polynucleotide having at least 97% sequence identity with a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 1 or its complementary nucleotide sequence, and encoding a protein having flavonoid 3′,5′-hydroxylase activity,(c) a polynucleotide encoding a protein comprising the amino acid sequence listed as SEQ ID NO: 2; and(d) a polynucleotide encoding a protein having an amino acid sequence with at least 97% sequence identity with the amino acid sequence listed as SEQ ID NO: 2, and having flavonoid 3′,5′-hydroxylase activity.2. A plasmid in which the polynucleotide according to is linked in a functional manner under the control of the cauliflower mosaic virus 35S promoter.3. A plasmid according to claim 2 , wherein the nos terminator is further linked in a functional manner.4Rosa hybridaAgrobacterium. A method for producing a transformed rose plant or its progeny having a delphinidin content of 80% or greater in the petals claim 2 , by transformation of a garden rose variety () by the method using a plasmid according to .5. The method according to claim 4 , wherein the delphinidin content in the petals is 85% or greater.6. The method according to claim 5 , wherein the delphinidin content in the petals is 90% or greater.7. The method according to claim 6 , wherein the delphinidin content in the petals is 95% or greater.8. A transformed rose plant or its progeny claim 4 , produced by the method according to .9Rosa hybridaAgrobacterium. A method for producing a transformed rose plant or its progeny having a delphinidin content of 80% or greater in the petals claim 3 , by transformation of a garden rose variety () by the method ...

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Номер записи: 37
07-03-2019 дата публикации

PRODUCTION OF GIBBERELLINS IN RECOMBINANT HOSTS

Номер: US20190071474A1
Автор: Carlsen Simon, Dyekjaer Jane Dannow, Hansen Esben Halkjaer, Naesby Michael, Ohler Nicholas, Rasmussen Nina Nicoline, Takos Adam Matthew
Принадлежит:

The invention relates to recombinant microorganisms and methods for producing compounds and precursors. 1. A recombinant host cell , comprising:(a) a recombinant gene encoding a first cytochrome P450 (P450) polypeptide; and/or(b) a recombinant gene encoding a 2-oxoglutarate-dependent dioxygenase (2-ODD) polypeptide and/or a second cytochrome P450 (P450) polypeptide;{'i': gibberellin', 'gibberellin, 'wherein the recombinant host cell is capable of producing a precursor and/or a compound.'}2. The recombinant host cell of claim 1 , wherein the gene encoding the first P450 polypeptide encodes a kaurenoic acid oxidase (KAO) polypeptide or a cytochrome P450 monooxygenase-1 (P450-1) polypeptide.3. The recombinant host cell of or claim 1 , wherein the gene encoding the first P450 polypeptide comprises:(a) a gene encoding a kaurenoic acid oxidase (KAO1) polypeptide;(b) a gene encoding a kaurenoic acid oxidase (KAO2) polypeptide;(c) a gene encoding a kaurenoic acid oxidase (KAO3) polypeptide;(d) a gene encoding a kaurenoic acid oxidase (KAO4) polypeptide;(e) a gene encoding a kaurenoic acid oxidase (KAO5) polypeptide;(f) a gene encoding a kaurenoic acid oxidase (KAO6) polypeptide;(g) a gene encoding a kaurenoic acid oxidase (KAO9) polypeptide;(h) a gene encoding a kaurenoic acid oxidase (KAO10) polypeptide;(i) a gene encoding a kaurenoic acid oxidase (KAO11) polypeptide;(j) a gene encoding a cytochrome P450 monooxygenase-2 (P450-2) polypeptide;(k) a gene encoding a cytochrome P450 monooxygenase-3 (P450-3) polypeptide;(l) a gene encoding a cytochrome P-450 BJ-1 (CYP112) polypeptide; and/or{'i': 'gibberellin', 'sub': '13ox', '(m) a gene encoding a A13-oxidase (GA) polypeptide.'}4. The recombinant host cell of claim 3 , wherein:(a) the KAO1 polypeptide comprises a KAO1 polypeptide having at least 50% sequence identity to the amino acid sequence set forth in SEQ ID NO:90;(b) the KAO2 polypeptide comprises a KAO2 polypeptide having at least 50% sequence identity to the amino acid ...

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Номер записи: 38
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: 39
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: 40
05-03-2020 дата публикации

Conversion of s-lignin compounds to useful intermediates

Номер: US20200071731A1
Автор: Christopher W. Johnson, Gregg Tyler BECKHAM, Sandra Fabienne NOTONIER
Принадлежит: Alliance for Sustainable Energy LLC

The present disclosure relates to a genetically modified microbial cell that includes a genetic modification resulting in the expression of a vanillate demethylase, where the microbial cell is capable of metabolizing at least one S-lignin decomposition molecule including at least one of syringate and/or 3-O-methyl gallate, and the genetically modified microbial cell is capable of producing gallate. In some embodiments of the present disclosure, the vanillate demethylase may include VanAB.

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Номер записи: 41
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: 42
30-03-2017 дата публикации

NITRIC OXIDE-MODULATING BONE-TARGETING COMPLEXES

Номер: US20170087221A1
Автор: Bangera Mahalaxmi Gita, Hyde Roderick A., Ishikawa Muriel Y., JR. Lowell L., Leuthardt Eric C., Wood
Принадлежит:

Compositions and methods are described herein for treating a bone disorder, including a bone-targeting complex including at least a portion of a nitric oxide synthase; a bone-targeting agent; and a linker coupling the at least a portion of the nitric oxide synthase to the bone-targeting agent. 1. A composition , comprising: at least a portion of a nitric oxide synthase;', 'a bone-targeting agent; and', 'a linker coupling the at least a portion of the nitric oxide synthase to the bone-targeting agent., 'a bone-targeting complex including'}2. (canceled)3. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase is conjugated to a first end of the linker and the bone-targeting agent is conjugated to a second end of the linker.4. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase comprises at least a portion of recombinant nitric oxide synthase.5. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase comprises at least a portion of endothelial nitric oxide synthase.6. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase comprises at least a portion of neuronal nitric oxide synthase.7. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase comprises at least a portion of inducible nitric oxide synthase.8. The composition of claim 1 , wherein the at least a portion of the nitric oxide synthase comprises a homodimer of the at least a portion of the nitric oxide synthase.9. The composition of claim 1 , wherein the bone-targeting agent comprises bisphosphonate.1012.-. (canceled)13. The composition of claim 1 , wherein the bone-targeting agent comprises a hydroxyapatite-binding polypeptide.14. The composition of claim 1 , wherein the bone-targeting agent comprises a bone morphogenetic protein.15. The composition of claim 1 , wherein the linker comprises a cleavable linker.16. The composition of claim 15 ...

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Номер записи: 43
05-05-2022 дата публикации

PROTECTING GROUP CHEMISTRY FOR CLEAN, REDUCTANT-FREE DYEING

Номер: US20220136024A1
Автор: BHAKTA Shyam Pravin, CERVANTES Bernardo, DUEBER John Eugene, FONG, HSU Tammy Melody, III Arthur Muir, Johnson, Jr. Terry Don, LATIMER Luke Nathaniel, PRATHURI Ramya Lakshmi, RUSS Zachary Nicholas
Принадлежит:

The present disclosure relates to the biosynthesis of indigoid dye precursors and their conversion to indigoid dyes. Specifically, the present disclosure relates to methods of using polypeptides to produce indigoid dye precursors from indole feed compounds, and the use of the indigoid dye precursors to produce indigoid dyes.

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Номер записи: 44
28-03-2019 дата публикации

AMPHIPATHIC PEPTIDE

Номер: US20190091344A1
Автор: Davison Ashley, McCarthy Helen, Robson Tracy, Zholobenko Aleksey
Принадлежит:

The present invention is directed to an amphipathic peptide and methods of using the amphipathic peptide for delivering small molecule agents to a cell. Ideally, the amphipathic cell penetrating peptide comprises less than approximately 50 amino acid residues with at least 6 arginine residues, at least 12 Alanine Residues, at least 6 leucine residues, optionally at least one cysteine residue, and at least two but no greater than three glutamic acids wherein the arginine residues are evenly distributed along the length of the peptide; and the peptide has a defined ratio of arginine to negatively charged amino acid residues and a defined ratio of hydrophilic amino acid residues to hydrophobic amino acid residues. The present invention is also directed to a nanoparticle and cell delivery system comprising the amphipathic cell penetrating peptide of the invention. The peptide, nanoparticle or cell delivery system of the invention may be used in therapy. For example, the peptide may be used as a therapeutic agent delivery system, in which the therapeutic agent may include nucleic acids or other small molecules. 1. An amphipathic cell penetrating peptide of less than approximately 50 amino acid residues comprising at least 6 arginine residues (R) , at least 12 alanine residues (A) , at least 6 leucine residues (L) , optionally at least one cysteine residue (C) , and at least two but no greater than three glutamic acids (E) wherein the arginine (R) residues are evenly distributed along the length of the peptide;the ratio of arginine (R) to negatively charged glutamic acid (E) residues is from at least 6:2 to 9:2, or at least 6:2 to 8:2; andthe ratio of hydrophilic amino acid residues to hydrophobic amino acid residues at pH 7 is at least 30:67 to 40:60, or at least 30:70 to 40:60; orwherein the peptide comprises or consists of WEARLARALARALARELARALARALRACEA (SEQ ID No. 4).2. The peptide according to wherein said peptide does not comprise glycine.3. The peptide according to ...

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Номер записи: 45
16-04-2015 дата публикации

METABOLICALLY ENGINEERED METHANOTROPHIC, PHOTOTROPHIC MICROORGANISMS

Номер: US20150104854A1
Автор: Kishore Ganesh M., Pakrasi Himadri, Singh Abhay
Принадлежит:

This disclosure relates to the engineering of phototrophic microorganisms for conversion of alkanes into higher-value products. Recombinant phototrophic organisms such as cyanobacteria can be engineered, optionally in a modular format, to express enzymes involved in converting methane to methanol, methanol to formaldehyde, formaldehyde to central metabolic pathway intermediates, and such intermediates to n-butanol. 1. A recombinant phototrophic microorganism , comprising one or more genes encoding a methane monooxygenase.2. The recombinant microorganism of claim 1 , wherein said MMO is a particulate MMO.3Methylococcus capsulatus. The recombinant microorganism of claim 1 , wherein said MMO comprises coding sequences for polypeptides having the amino acid sequences of Bath PmoA claim 1 , PmoB claim 1 , and PmoC.4. The recombinant microorganism of claim 1 , wherein said MMO is a soluble MMO.5. The recombinant microorganism of claim 4 , wherein said sMMO comprises one or more recombinant genes encoding the following polypeptides: an MmoX polypeptide; an MmoY polypeptide claim 4 , an MmoB polypeptide claim 4 , an MmoZ polypeptide claim 4 , an MmoD claim 4 , and an MmoC polypeptide.6. The recombinant microorganism of claim 1 , wherein expression of said one or more genes in said microorganism results in the production of methanol; ethanol; propanol claim 1 , or n-butanol claim 1 , when said microorganism is grown in the presence of light and Oin a medium comprising methane claim 1 , ethane claim 1 , propane or butane claim 1 , respectively.7. The recombinant microorganism of claim 1 , further comprising:i) a methanol dehydrogenase or a human class I alcohol dehydrogenase;ii) a hexulose-6-phosphate synthase and a 6-phosphate-3-hexuloisomerase; andiii) recombinant genes encoding an acetyl-CoA acetyltransferase polypeptide; a 3-hydroxybutyryl-CoA dehydrogenase polypeptide; a 3-hydroxybutyryl-CoA dehydratase (crotonase) polypeptide; an aldehyde/alcohol dehydrogenase ...

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Номер записи: 46
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: 47
26-03-2020 дата публикации

METHYLOTROPHIC MICROORGANISMS EXPRESSING SOLUBLE METHANE MONOOXYGENASE PROTEINS

Номер: US20200095611A1
Автор: FLEURY Samantha Therese, GALAZKA Jonathan Matthew, Rahman Asif
Принадлежит:

Methylotrophic microorganisms, particularly methylotrophic yeasts and more particularly , which exhibit the ability to oxidize methane to methanol. Methods of making such microorganisms and DNA constructs for making such microorganisms. Such methylotrophic microorganisms are genetically transformed to exhibit the oxidizing activity of a soluble methane monooxygenase of a methanotrophic bacterium. Such transformed methylotrophic microorganisms contain at least three methane monooxygenase hydroxylase (MMOH) protein subunits of a methanotrophic bacterium: MMOH alpha, MMOH beta and MMOH gamma and a methane monooxygenase reductase (MMOR) of a methanotrophic bacterium. 1. A methylotrophic microorganism comprising three methane monooxygenase hydroxylase (MMOH) protein subunits of a methanotrophic bacterium: MMOH alpha , MMOH beta and MMOH gamma and methane monooxygenase reductase (MMOR) of a methanotrophic bacterium , wherein the methane monooxygenase (MMO) is a soluble MMO and wherein the microorganism exhibits oxidation of methane to methanol.2. The methylotrophic microorganism of claim 1 , which is a methylotropic yeast.3Pichia.. The methylotrophic microorganism of claim 1 , which is a methylotropic strain of the genus4Pichia pastoris.. The methylotropic microorganism of claim 1 , which is5Methylosinus trichosporiumMethylococcus capsulatus.. The methylotropic microorganism of claim 1 , wherein the methanotrophic bacterium is a strain of or6. The methylotrophic microorganism of claim 1 , which converts methane to microbial products.7. The methylotrophic microorganism of claim 1 , which at least in part employs methane as a carbon source.8. The methylotropic microorganism of claim 1 , wherein the nucleotide coding sequences of the three MMOH protein subunits are expressed in the microorganism as a single transcript.9. The methylotropic microorganism of claim 8 , wherein in the single transcript the nucleotide coding sequences of MMOH protein subunits are separated by ...

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Номер записи: 48
21-04-2016 дата публикации

FUSION PROTEINS, RECOMBINANT BACTERIA, AND METHODS FOR USING RECOMBINANT BACTERIA

Номер: US20160108096A1
Автор: Siegel Ashley, Thompson Brian
Принадлежит:

Fusion proteins containing a targeting sequence, an exosporium protein, or an exosporium protein fragment that targets the fusion protein to the exosporium of a family member are provided. Recombinant family members expressing such fusion proteins are also provided. Genetically inactivated family members and recombinant family members that overexpress exosporium proteins are also provided. Seeds coated with the recombinant family members and methods for using the recombinant family members (e.g., for stimulating plant growth) are also provided. Various modifications of the recombinant family members that express the fusion proteins are further provided. Fusion proteins comprising a spore coat protein and a protein or peptide of interest, recombinant bacteria that express such fusion proteins, seeds coated with such recombinant bacteria, and methods for using such recombinant bacteria (e.g., for stimulating plant growth) are also provided. 1. A fusion protein comprising at least one protein or peptide of interest and:(1) a targeting sequence comprising amino acids 1-30 of SEQ ID NO: 59;(2) a targeting sequence comprising SEQ ID NO: 59;(3) an exosporium protein comprising an amino acid sequence having at least 85% identity with SEQ ID NO: 60;(4) a targeting sequence comprising amino acids 2-30 of SEQ ID NO: 59;(5) a targeting sequence comprising amino acids 4-30 of SEQ ID NO: 59;(6) a targeting sequence comprising amino acids 6-30 of SEQ ID NO: 59;(7) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 61;(8) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 61;(9) a targeting sequence comprising SEQ ID NO: 61;(10) an exosporium protein comprising an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 62;(11) a targeting sequence comprising amino acids 2-33 of SEQ ID NO: 61;(12) a targeting sequence comprising amino acids 5-33 of SEQ ID NO: 61;(13) a targeting sequence comprising amino acids 10-33 of SEQ ID NO: 61;(14) a ...

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Номер записи: 49
02-06-2022 дата публикации

NOVEL ENZYME NANOPARTICLES HAVING METHANE OXIDATION ACTIVITY

Номер: US20220170051A1
Автор: Kim Hyun Jin, LEE Jeewon
Принадлежит: KOREA UNIVERSITY RESEARCH AND BUSINESS FOUNDATION

The present invention relates to novel enzyme nanoparticles capable of converting methane into methanol, in which key active sites of methane-oxidizing bacteria are fused with each other and expressed on a protein that can be self-assembled in cells to form nanoparticles, and specifically to enzyme nanoparticles including a protein having methane monooxygenase (MMO) activity and active sites of the methane oxidase, a method for production thereof, a recombinant microorganism into which a nucleic acid encoding the protein and the active site of the methane oxidase is introduced, and immobilized enzyme nanoparticles including the enzyme nanoparticles loaded on a carrier. 1. An enzyme nanoparticle having methane monooxygenase (MMO) (“methane oxidase”) activity , wherein a first active site and a second active site of the methane oxidase are fused with each other and expressed on a surface of a protein adapted to be self-assembled in cells to form nanoparticles.2Escherichia coli. The enzyme nanoparticle according to claim 1 , wherein the protein is ferritin claim 1 , ferritin-like protein claim 1 , magnetosome constituent protein claim 1 , DNA binding protein claim 1 , human α-tocopherol transfer protein claim 1 , human PERK chaperone claim 1 , hepatitis B virus capsid protein claim 1 , proteasome claim 1 , or tobacco mosaic virus coat protein.3. The enzyme nanoparticle according to claim 1 , wherein the protein is a ferritin heavy-chain protein.4. The enzyme nanoparticle according to claim 1 , wherein the first active site and the second active site of the methane oxidase are selected from the group consisting of pmoB1 or a variant thereof and pmoB2 or a variant thereof claim 1 , and wherein the first active site and the second active site are different from each other.5. The enzyme nanoparticle according to claim 1 , wherein the first active site of the methane oxidase includes the sequence of SEQ ID NO: 2 claim 1 , and the second active site includes the sequence of ...

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Номер записи: 50
23-04-2015 дата публикации

Process for the production of methyl methacrylate

Номер: US20150112037A1
Автор: Adrianus Johannes Jozef Straathof, David William Johnson, Graham Ronald Eastham, Marco Wilhemus Fraaije, Remko Tsjibbe Winter
Принадлежит: Lucite International UK Ltd

The invention includes a process of producing methyl methacrylate or derivatives thereof is described. The process includes the steps of converting 2-butanone to methyl propionate using a Baeyer-Villiger monooxygenase, and treating the methyl propionate produced to obtain methyl methacrylate or derivatives thereof. A method of preparing polymers or copolymers of methyl methacrylate or its derivatives is also described.

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Номер записи: 51
27-04-2017 дата публикации

BIOFILMS, COMPONENTS AND METHODS OF USE TO REDUCE BIOFOULING AND CONTAMINATION

Номер: US20170112136A1
Автор: Kumar Manish, Wood Thammajun L., Wood Thomas K.
Принадлежит:

Biofilms are provided which are capable of regulating their own thickness, reducing contamination and preventing biofouling. Constructs are introduced into bacteria that comprise nucleic acid molecules encoding an autoinducer synthase polypeptide, a transcriptional regulator and a biofilm dispersal protein. Nucleic acid molecules may also be introduced which encode a nitric oxide synthase, an epoxide hydrolase, or both. Biofilms of the bacteria may be used to reduce biofouling and contamination of a surface. 1. A method of reducing biofouling of a surface , said method comprising: i. a nucleic acid molecule encoding an autoinducer synthase polypeptide;', 'ii. a nucleic acid molecule encoding a transcriptional regulator, capable of being activated by said autoinducer synthase polypeptide; and', 'iii. a nucleic acid molecule encoding a biofilm dispersal protein; and, 'a) introducing into at least one bacterial cell a quorum sensing nucleic acid construct, comprising,'}b) producing a biofilm of said bacterial cells, wherein said quorum sensing nucleic acid construct reduces the thickness of said biofilm compared to biofilm not comprising said quorum sensing nucleic acid construct.2. The method of claim 1 , wherein said autoinducer synthase comprises LasI and said transcriptional regulator comprises LasR.3. The method of claim 1 , wherein said biofilm dispersal protein comprises BdcA4. The method claim 2 , wherein said biofilm dispersal protein comprises BdcA.5. The method of claim 1 , further comprising introducing into said at least one bacterial cell a nucleic acid sequence encoding nitric oxide synthase.6. The method of claim 1 , further comprising introducing into said bacterial cell a nucleic acid molecule encoding epoxide hydrolase.7. The method of claim 2 , above claim 2 , further comprising a nucleic acid encoding epoxide hydrolase.8. The method of claim 4 , further comprising introducing into said bacterial a nucleic acid molecule encoding epoxide hydrolase.9. ...

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Номер записи: 52
18-04-2019 дата публикации

IN VITRO METHOD FOR IDENTIFYING THORACIC AORTIC ANEURYSMS (TAA) IN A SUBJECT

Номер: US20190112582A1
Автор: CAMPANERO GARCÍA Miguel Ramón, MÉNDEZ-BARBERO Nerea, OLLER PEDROSA Jorge, REDONDO MOYA Juan Miguel
Принадлежит:

The present invention refers to an In vitro method for screening for subjects at risk of developing thoracic aortic aneurysm (TAA) or a disease causing TAA comprising: (a) measuring the expression pattern or level of at least A Disintegrin And Metalloproteinase with Thrombospondin Motifs 1 (ADAMTS1) obtained from an isolated biological sample of the subjects to be screened; and (b) comparing said expression pattern or level of at least ADAMTS1 of the subjects to be screened with an already established expression pattern or level, wherein reduced expression of at least ADAMTS1 is indicative of a thoracic aortic aneurysm (TAA). 1. A method for the prophylactic or therapeutic treatment of a thoracic aortic aneurysm (TAA) in a patient , comprising administering a composition comprising an iNOS inhibitor to the patient.2. The method of claim 1 , wherein said iNOS inhibitor is capable of binding in the guanidine site of the L-arginine ligand region of the human NOS2 structure and inhibiting the human NOS2 isoform in the presence of L-arginine.4. The method of claim 1 , wherein the iNOS inhibitor is a selective iNOS inhibitor.5. The method of claim 1 , wherein the iNOS inhibitor is selected from the group consisting of 1400 W claim 1 , L-NAME claim 1 , GW274150 claim 1 , GW273629 claim 1 , Aminoguanidine (AG) hydrocloride claim 1 , L-NIL and clotrimazale.6. The method of claim 1 , wherein the patient is suffering from a disease selected from the group consisting of: bicuspid aortic valve; a syndromic thoracic aortic aneurysm (TAA) such as Marfan Syndrome claim 1 , vascular Ehlers Danlos claim 1 , Loeys Dietz Syndrome (Types 1 and 2) claim 1 , and Familial thoracic aortic aneurysm and dissection (familial TAAD) claim 1 , a non-syndromic TAA or any other disease associated with an aorthopathy triggered by Adamts1 deficiency.7. The method of claim 5 , wherein the patient is suffering from a disease selected from the group consisting of: a syndromic thoracic aortic aneurysm ( ...

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Номер записи: 53
09-04-2020 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20200109376A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 136 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 136 , selected from the group consisting of 75 , 79 , 82 , 99 , 110 , 166 , 172 , 208 , 216 , 273 , 324 , 364 , 395 , 412 , 491 , 503 , and 504.3. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 136.5. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: 136 selected from the group consisting of a glycine at position 143 claim 1 , glycine at position 278 claim 1 , an arginine at position 326 claim 1 , and ...

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Номер записи: 54
25-04-2019 дата публикации

TRANSFORMANT, AND METHOD FOR PRODUCING PROTOCATECHUIC ACID OR SALT THEREOF USING SAME

Номер: US20190119664A1
Автор: HIRAGA Kazumi, INUI MASAYUKI, Kogure Takahisa, SUDA Masako
Принадлежит:

Provided is a microorganism that is able to efficiently produce protocatechuic acid or a salt thereof by using a saccharide as a raw material, and a method of efficiently producing protocatechuic acid or a salt thereof by using the microorganism. 1. A transformant having protocatechuic acid producing ability ,wherein the transformant is subjected to modifications (A), (B), and (C) below:(A) enhancement of 3-dehydroshikimate dehydratase activity;(B) enhancement of chorismate pyruvate lyase activity; and(C) enhancement of 4-hydroxybenzoate hydroxylase activity.2. The transformant according to claim 1 ,{'i': Corynebacterium', 'Rhodococcus', 'Bacillus', 'Rhodopseudomonas', 'Alteromonas', 'Marinobacter', 'Methylobacterium', 'Pantoea', 'Neurospora', 'Aspergillus., 'wherein the enhancement of 3-dehydroshikimate dehydratase activity is achieved by introducing, into a host, a gene that encodes an enzyme having 3-dehydroshikimate dehydratase activity, the gene being derived from a microorganism belonging to the genus , the genus , the genus , the genus , the genus , the genus , the genus , the genus , the genus , or the genus'}3. The transformant according to claim 2 ,{'i': Corynebacterium glutamicum, Corynebacterium halotolerans, Corynebacterium casei, Corynebacterium efficiens, Aspergillus niger', 'Aspergillus oryzae., 'wherein the gene that encodes the enzyme having 3-dehydroshikimate dehydratase activity is a gene of , or'}4. The transformant according to claim 2 ,wherein the gene that encodes the enzyme having 3-dehydroshikimate dehydratase activity is encoded by a DNA of (a) or (b) below:(a) a DNA which consists of a base sequence of SEQ ID NO. 7, 134, 135, 145, 147, or 149; or(b) a DNA which consists of a base sequence having 90% or more of identity with a base sequence of SEQ ID NO. 7, 134, 135, 145, 147, or 149, the DNA encoding a polypeptide having 3-dehydroshikimate dehydratase activity.5. The transformant according to claim 1 ,{'i': Providencia', 'Cronobacter., ' ...

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Номер записи: 55
12-05-2016 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20160130566A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide sequence encoding a polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 95% sequence identity to SEQ ID NO:1362. The polynucleotide sequence of claim 1 , wherein said encoded polypeptide is capable of converting the acid substrate compound (1b) to compound (2b) (R-enantiomer) or its opposite enantiomer compound (S-enantiomer) with at least 2-fold improved activity relative to the wild-type polypeptide of SEQ ID NO: 2.3. The polynucleotide of claim 1 , wherein said encoded polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 selected from: X246A claim 1 , E claim 1 , G claim 1 , I claim 1 , L claim 1 , P claim 1 , S claim 1 , T claim 1 , and V.4. The polynucleotide of claim 1 , wherein said encoded polypeptide is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% ee.5. The polynucleotide of claim 1 , wherein said encoded polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 selected from: X143G; X278G; X326R; and X490L.6. The polynucleotide of claim 1 , wherein said encoded polypeptide further comprises at least the following amino acid differences relative to SEQ ID NO: 2: X277I; X278A claim 1 , or G; X280T or Y; X281I; X326R; and X490L or X490Q.8. The ...

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Номер записи: 56
17-05-2018 дата публикации

METHODS FOR PRODUCTION OF OXYGENATED TERPENES

Номер: US20180135081A1
Автор: Ghosh Souvik, KUMARAN Ajikumar Parayil, LI Liwei, Lim Chin Giaw, Pirie Christopher, Qualley Anthony
Принадлежит:

The present invention relates to methods for producing oxygenated terpenoids. Polynucleotides, derivative enzymes, and host cells for use in such methods are also provided. 1Stevia rebaudiana. A method for making an oxygenated product of valencene , comprising , contacting the valencene with Kaurene Oxidase (SrKO) or derivative thereof having valencene oxidizing activity , and recovering an oxygenated product that is predominantly nootkatone.2. (canceled)3. (canceled)4. The method of claim 1 , wherein the SrKO derivative comprises an amino acid sequence having at least 70% sequence identity to SrKO (SEQ ID NO: 37 claim 1 , 38 claim 1 , or 55) claim 1 , and has valencene oxidase activity.5. The method of claim 1 , wherein one or more mutations relative to SrKO (SEQ ID NO: 37 claim 1 , 38 claim 1 , or 55) are selected from Table 2 or Table 6.6. (canceled)7E. coli. The method of claim 1 , wherein the SrKO is a derivative having a deletion of at least a portion of its N-terminal transmembrane region claim 1 , and the addition of an inner membrane transmembrane domain from yhcB or derivative thereof8. (canceled)9. (canceled)10. The method of claim 1 , wherein the SrKO is a derivative having one or more mutations at positions selected from 46 claim 1 , 76 claim 1 , 94 claim 1 , 131 claim 1 , 231 claim 1 , 284 claim 1 , 383 claim 1 , 390 claim 1 , 400 claim 1 , 444 claim 1 , 468 claim 1 , 488 and 499 relative to SEQ ID NO:37.11. The method of claim 10 , wherein the SrKO is a derivative having one or more mutations selected from R76K claim 10 , M94V claim 10 , T131Q claim 10 , F231L claim 10 , H284Q claim 10 , R383K claim 10 , I390L claim 10 , T468I claim 10 , and T499N relative to SEQ ID NO:37.12. The method of claim 1 , wherein the SrKO is a derivative comprising an amino acid sequence selected from SEQ ID NOS:55-61 claim 1 , 104 claim 1 , and 105 or an amino acid sequence having from one to twenty mutations relative to a sequence selected from SEQ ID NOS: 55-61 claim 1 , ...

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Номер записи: 57
18-05-2017 дата публикации

BACTERIA ENGINEERED FOR CONVERSION OF ETHYLENE TO N-BUTANOL

Номер: US20170137846A1
Автор: ATSUMI Shota, CARLIN D. Alexander, DESAI Shuchi H., KORYAKINA Irina, RODRIGUEZ Gabriel M., SIEGEL Justin B., TASHIRO Yohei, TONEY Michael D.
Принадлежит: The Regents of the University of California

The present disclosure provides recombinant bacteria with elevated production of ethanol and/or n-butanol from ethylene. Methods for the production of the recombinant bacteria, as well as for use thereof for production of ethanol and/or n-butanol are also provided. 1. A bacterium comprising a recombinant polynucleotide encoding an ethylene hydratase (EH) , wherein expression of the EH results in an increase in production of ethanol as compared to a corresponding bacterium lacking the recombinant polynucleotide.2. The bacterium of claim 1 , wherein the EH is an oleate hydratase.3Lysinibacillus fusiformis. The bacterium of claim 2 , wherein the oleate hydratase is a oleate hydratase.4. The bacterium of claim 1 , wherein the EH is a 2-haloacrylate hydratase.5Pseudomonas. The bacterium of claim 4 , wherein the 2-haloacrylate hydratase is a species 2-haloacrylate hydratase.6. The bacterium of claim 1 , wherein the EH is a kievitone hydratase.7Fusarium solani. The bacterium of claim 6 , wherein the kievitone hydratase is a kievitone hydratase.8. A bacterium comprising a recombinant polynucleotide encoding an alkene monooxygenase (AMO) and an ethylene oxide reductase (EOR) claim 6 , wherein expression of the AMO and the EOR results in an increase in production of ethanol as compared to a corresponding bacterium lacking the recombinant polynucleotide.9. The bacterium of claim 8 , wherein the AMO is a toluene monooxygenase.10Pseudomonas mendocina. The bacterium of claim 9 , wherein the toluene monooxygenase is a toluene monooxygenase.11Burkholderia cepacia. The bacterium of claim 9 , wherein the toluene monooxygenase is a toluene monooxygenase.12. The bacterium of claim 8 , wherein the EOR is an NAD dependent formate dehydrogenase.13. The bacterium of claim 8 , wherein the recombinant polynucleotide comprises a first polynucleotide encoding the alkene monooxygenase (AMO) and a second polynucleotide encoding the ethylene oxide reductase (EOR).1421-. (canceled)22. A bacterium ...

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Номер записи: 58
25-05-2017 дата публикации

MICROBIAL PRODUCTION OF MUCONIC ACID AND SALICYLIC ACID

Номер: US20170145447A1
Автор: LIN Yuheng, Yan Yajun
Принадлежит: University of Georgia Research Foundation, Inc.

The invention provides a recombinant microorganism that has been genetically engineered to contain metabolic pathway for the production of muconic acid from a salicylic acid intermediate. The genetically engineered metabolic pathway comprises both biosynthetic and biodegradative elements. 1. A genetically engineered microorganism comprising a genetically engineered metabolic pathway for the production of muconic acid from a salicylic acid intermediate , said pathways comprising:a plurality of enzymes associated with the biosynthesis of salicylic acid, said plurality of enzymes comprising an isochorismate synthase (ICS) and an isochorismate pyruvate lyase (IPL); anda plurality of enzymes associated with the conversion of salicylic acid to muconic acid, said plurality of enzymes comprising a salicylate 1-monoxygenase (SMO) and a catechol 1,2-dioxygenase (CDO).24.-. (Canceled)5. The genetically engineered microorganism of wherein the genetically engineered metabolic pathway further comprises at least one enzyme associated with enhanced chorismate availability.6. The genetically engineered microorganism of wherein the enzyme associated with enhanced chorismate availability is selected from the group consisting of a shikimate kinase claim 5 , a phosphoenolpyruvate synthase claim 5 , a transketolase claim 5 , a 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase claim 5 , a 3-dehydroquinate synthase claim 5 , a 3-dehydroquinate dehydratase claim 5 , a shikimate dehydrogenase claim 5 , a 3-phosphoshikimate-1-carboxyvinyltransferase claim 5 , and a chorismate synthase.7. The genetically engineered microorganism of comprising:a first module expressing an isochorismate synthesis (ICS) and an isochorismate pyruvate lyase (IPL); anda second module expressing a salicylate 1-monoxygenase (SMO) and a catechol 1,2-dioxygenase (CDO).8. The genetically engineered microorganism of wherein the first and second modules are present on different plasmids.9. The genetically engineered ...

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Номер записи: 59
07-05-2020 дата публикации

Bradyrhizobium monooxygenase and use thereof for preparation of chiral sulfoxide

Номер: US20200140830A1
Автор: Feng Liu, Guoqiang Lin, Huilei YU, JIANG Pan, Jianhe Xu, Qian Zhao, Yan Zhang
Принадлежит: EAST CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY, Jiangsu Aosaikang Pharmaceutical Co Ltd

A Bradyrhizobium monooxygenase, a gene for encoding the monooxygenase, a recombinant expression vector comprising the gene and a recombinant transformant, a method of preparing the monooxygenase by the recombinant expression transformant, and a method of preparing an optically pure chiral sulfoxide by the monooxygenase, in particular to a method of preparing prazole drugs by means of catalyzing the asymmetric oxidation of thioether, a prazole precursor. As compared with other methods of preparing an optically pure sulfoxide, the product produced by the monooxygenase of the present invention as a catalyst has high optical purity, avoids the generation of the byproduct sulfone, and has advantages of mild reaction conditions, simple and convenient operations, easy amplification, etc.

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Номер записи: 60
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: 61
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: 62
23-05-2019 дата публикации

Mycobacterium Neoaurum-Derived Steroid C27-monooxygenase and Application Thereof

Номер: US20190153404A1
Автор: RAO Zhiming, Shao Minglong, XU Meijuan, YANG Taowei, Zhang Xian
Принадлежит:

The present invention discloses a -derived steroid C27-monooxygenase and an application thereof, which belong to the technical fields of genetic engineering and enzyme engineering. By the method of gene knockout and intensive expression, the present invention screens out three isoenzymes of a key enzyme SMO in the process of degrading sterol side chains from . The three isoenzymes are intensively expressed respectively in the for the high yield of androsta-1,4-diene-3,17-dione (ADD), the yield of ADD is increased remarkably, wherein the effect of SMO2 is most remarkable. By overexpressing SMO2, the final ADD yield is increased from 5.2 g·L.to 7.3 g·L.. The present invention provides a helpful guidance for the industrialization of the microbial fermentation method for increasing the ADD yield. 1Mycobacterium neoaurumMycobacterium neoaurum. Recombinant with an increased yield of ADD , wherein the recombinant comprises an overexpression of a steroid C27-monooxygenase gene; and the steroid C27-monooxygenase gene is selected from the group consisting of Smo1 gene , Smo2 gene and Smo3 gene.2Mycobacteriurn neoaurum. The recombinant according to claim 1 , characterized in that the amino acid sequences of the steroid C27-monooxygenase genes Smo1 claim 1 , Smo2 and Smo3 are set forth respectively in SEQ ID NO.4 claim 1 , SEQ ID NO.5 and SEQ ID NO.6.3Mycobacterium neoaurumMycobacterium neoaurum. The recombinant according to claim 1 , characterized in that with regard to the overexpression claim 1 , the recombinant comprises a recombinant plasmid comprising the steroid C27-monooxygenase gene.4Mycobacterium neoaurumMycobacterium neoaurum. A method for increasing the yield of ADD by overexpressing a steroid C27-monooxygenase gene claim 1 , comprising using the recombinant of as a production strain; overexpressing the steroid C27-monooxygenase gene in the recombinant ; and fermentatively producing ADD.55. The method according to claim 4 , characterized in that the amino acid ...

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Номер записи: 63
08-06-2017 дата публикации

CELL-FREE SYSTEM FOR CONVERTING METHANE INTO FUEL AND CHEMICAL COMPOUNDS

Номер: US20170159058A1
Автор: Blake William Jeremy, Swartz James R.
Принадлежит: GreenLight Biosciences, Inc.

The present disclosure relates, in some aspects, to cell-free methods and systems for large-scale conversion of methane to isobutanol, comprising combining, in a bioreactor at elevated pressure, methane, oxygen, and cell lysates containing methane monooxygenase, methanol dehydrogenase, and enzymes that catalyze the conversion of formaldehyde to isobutanol, to form a cell-free reaction mixture, and incubating under suitable conditions the cell-free reaction to convert methane to isobutanol. 175-. (canceled)76. A composition comprising at least two cell lysates and the following enzymes: a methane monooxygenase , a methanol dehydrogenase , a hexulose-6-phosphate synthase , a 6-phospho-3-hexuloisomerase , a 6-phosphofructokinase , a fructose bisphosphate aldolase , a triose phosphate isomerase , a transketolase , a ribose-5-phosphate isomerase or a ribulose-5-phosphate 3-epimerase , a glyceraldehyde 3-phosphate dehydrogenase , a phosphoglycerate kinase , a phosphoglycerate mutase , an enolase , and a pyruvate kinase , wherein at least one of the cell lysates is obtained from recombinant cells that overexpress at least one of the foregoing enzymes.77. The composition of claim 76 , wherein(i) the methane monooxygenase has EC number 1.14.13.25 or 1.14.18.3,(ii) the methanol dehydrogenase has EC number 1.1.1.244, 1.1.2.7, or 1.1.99.37,(iii) the hexulose-6-phosphate synthase has EC number 4.1.2.43,(iv) the 6-phospho-3-hexuloisomerase has EC number 5.3.1.27,(v) the 6-phosphofructokinase has EC number 2.7.1.11,(vi) the fructose bisphosphate aldolase has EC number 4.1.2.13,(vii) the triose phosphate isomerase has EC number 5.3.1.1,(viii) the transketolase has EC number 2.2.1.1,(xi) the ribulose-5-phosphate 3-isomerase has EC number 5.3.1.6, or the ribulose-5-phosphate 3-epimerase has EC number 5.1.3.1,(xii) the glyceraldehyde 3-phosphate dehydrogenase has EC number 1.2.1.12,(xiii) the phosphoglycerate kinase has EC number 2.7.2.3,(xiv) the phosphoglycerate mutase has EC number ...

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Номер записи: 64
14-05-2020 дата публикации

METHOD FOR UTILIZING ENGINEERED DENDRITIC CELLS TO INDUCE GUT-HOMING REGULATORY T CELLS AND TREAT GUT INFLAMMATION

Номер: US20200149121A1
Автор: Baylink David J., Lau K.-H. William, Tang Xiaolei, Walter Michael
Принадлежит:

Gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme. A method for treating one or more than one inflammation-related condition or disease, the method comprising administering gene-modified, lymphoid-tissue-homing dendritic cells that comprise a 1-alpha-hydroxylase gene and a retinaldehyde dehydrogenase 2 gene, where the 1-alpha-hydroxylase gene is expressed to produce functional 1-alpha-hydroxylase enzyme and the retinaldehyde dehydrogenase 2 gene is expressed to produce functional retinaldehyde dehydrogenase 2 gene enzyme. 1. A gene-modified dendritic cell , the gene-modified dendritic cell comprising a cytochrome p450 family 27 subfamily B polypeptide 1 (CYP27B1) gene encoding 1-alpha-hydroxylase and an aldehyde dehydrogenase 1 family member A2 (ALDH1a2) gene encoding retinaldehyde dehydrogenase 2 (RALDH2).2. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is engineered to constitutively overexpress the 1-alpha-hydroxylase.3. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is engineered to constitutively overexpress the RALDH2.4. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is engineered to constitutively overexpress both the 1-alpha-hydroxylase and the RALDH2.5. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is a bone marrow-derived dendritic cell.6. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell is a bone marrow-derived dendritic cell.7. The gene-modified dendritic cell of claim 1 , wherein the gene-modified dendritic cell contains a bicistronic expression vector with a ...

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Номер записи: 65
29-09-2022 дата публикации

Multi-Enzyme Nanoparticle-Assisted Stable Isotope Incorporation Into Small Molecules by Channeling

Номер: US20220307004A1
Автор: Breger Joyce C., Ellis Gregory A., Malone Kirk, Medintz Igor L., Scrutton Nigel
Принадлежит:

Multi-enzyme systems attached to nanoparticles are effective to efficiently and controllably incorporate stable isotopes (such as deuterium) during the synthesis of small molecules. In one example, deuterium is incorporated into (+)-dihydrocarvide using a cascade involving the enzymes (a) pentaerythritol tetranitrate reductase (PETNR) and (b) flavin-dependent cyclohexanone monooxygenase triple variant F249A/F280A/F435A (CHMO). 1. A method of incorporating a radioisotope into a product molecule comprises:providing a nanoparticle attached to a plurality of enzymes configured as an enzymatic cascade such that a product of a first enzyme is a substrate of the second enzyme and so forth;providing a radioisotope source and a source substrate to the enzymatic cascade; andallowing the enzymatic cascade to act on the radioisotope source and the source substrate, thereby transferring a radioisotope from the radioisotope source into a product molecule.2. The method of claim 1 , wherein the radioisotope source is provided in the form of a solvent and/or one or more cofactors of the enzymes.3. The method of claim 1 , wherein the radioisotope source is a deuterium source.4. The method of claim 3 , wherein the deuterium source is selected from the group consisting of as deuterated water claim 3 , deuterated nicotinamide adenine dinucleotide phosphate (NADH-11) claim 3 , deuterated reduced nicotinamide adenine dinucleotide (NADH-11) claim 3 , and combinations thereof.5. The method of claim 1 , wherein the enzymatic cascade comprises one or more reductase enzymes effective to utilize at least one of the radioisotope sources.6. The method of claim 5 , wherein said one or more reductase enzymes comprise pentaerythritol tetranitrate reductase (PETNR) and/or carboxylic acid reductase (CAR).7. The method of claim 1 , whereinthe enzymatic cascade comprises pentaerythritol tetranitrate reductase and a cyclohexanone monooxygenase;and the product molecule is dihydrocarvide.8. The method of ...

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Номер записи: 66
30-05-2019 дата публикации

TRANSGENIC PLANTS WITH INCREASED PHOTOSYNTHESIS EFFICIENCY AND GROWTH

Номер: US20190161765A1
Автор: Gabilly Stephane T., Glowacka Katarzyna, Kromdijk Johannes, Leonelli Laurie, Long Stephen P., Niyogi Krishna K.
Принадлежит:

The present disclosure provides a transgenic plant comprising one or more nucleotide sequences encoding polypeptides selected from photosystem II subunit S (PsbS), zeaxanthin epoxidase (ZEP), and violaxanthin de-epoxidase (VDE), operably linked to at least one expression control sequence. Expression vectors for making transgenic plants, and methods for increasing biomass production and/or carbon fixation and/or growth in a plant comprising increasing expression of at least one of PsbS, ZEP and VDE polypeptides are also provided. 1. A transgenic plant comprising one or more heterologous nucleotide sequences encodingphotosystem II subunit S (PsbS), zeaxanthin epoxidase (ZEP), and violaxanthin de-epoxidase (VDE);PsbS and ZEP; orZEP and VDE, operably linked to at least one expression control sequence.2Arabidopsis thaliana.. The transgenic plant of claim 1 , wherein the one or more heterologous nucleotide sequences are derived from a dicot or wherein the one or more heterologous nucleotide sequences are derived from3. (canceled)4. The transgenic plant of claim 1 , comprising one or more heterologous nucleotide sequences encoding PsbS claim 1 , ZEP and VDE.5. The transgenic plant of claim 1 , wherein the transcript levels of any of VDE claim 1 , PsbS or ZEP are increased as compared to a control plant.6. The transgenic plant of claim 1 , wherein:PsbS is encoded by the nucleotide sequence of SEQ ID NO: 1, ZEP is encoded by the nucleotide sequence of SEQ ID NO:2, and VDE is encoded by the nucleotide sequence of SEQ ID NO: 3;PsbS is encoded by a nucleotide sequence having at least 90% identity to SEQ ID NO: 1, ZEP is encoded by a nucleotide sequence having at least 90% identity to SEQ ID NO:2, and VDE is encoded by a nucleotide sequence having at least 90% identity to SEQ ID NO: 3;PsbS is encoded by a nucleotide sequence having at least 70% identity to SEQ ID NO: 1, ZEP is encoded by the nucleotide sequence having at least 70% identity to SEQ ID NO:2, and VDE is encoded by ...

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Номер записи: 67
01-07-2021 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20210198638A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 2 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 2 , selected from the group consisting of 280 , 322 , 325 , 426 , 430 , 432 , 435 , and 532.3. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 2 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 2.5. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: 2 selected from the group consisting of a glycine at position ...

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Номер записи: 68
04-06-2020 дата публикации

IMPROVED METHANE MONOOXYGENASE ENZYMES

Номер: US20200172881A1
Автор: CLARKE Elizabeth Jane, Greenfield Derek Lorin, Helman Noah Charles, Jones Stephanie Rhianon, Zhu Baolong
Принадлежит:

Improved soluble methane monooxygenases and soluble methane monooxygenase systems are provided. 1. A mutant soluble diiron monooxygenase system comprising at least one mutant subunit , wherein the at least one mutant subunit comprises one or more mutations , such that the activity of the mutant soluble diiron monooxygenase system is greater than the activity of a wild-type soluble diiron monooxygenase system.2. The mutant soluble diiron monooxygenase system of claim 1 , comprising a mutant hydroxylase alpha subunit wherein the mutant hydroxylase alpha subunit comprises one or more mutations at one or more residue position(s): 3 claim 1 , 14 claim 1 , 25 claim 1 , 26 claim 1 , 32 claim 1 , 33 claim 1 , 35 claim 1 , 42 claim 1 , 44 claim 1 , 53 claim 1 , 54 claim 1 , 55 claim 1 , 56 claim 1 , 57 claim 1 , 61 claim 1 , 130 claim 1 , 135 claim 1 , 154 claim 1 , 162 claim 1 , 163 claim 1 , 168 claim 1 , 216 claim 1 , 251 claim 1 , 262 claim 1 , 263 claim 1 , 264 claim 1 , 277 claim 1 , 316 claim 1 , 322 claim 1 , 330 claim 1 , 347 claim 1 , 356 claim 1 , 374 claim 1 , 375 claim 1 , 381 claim 1 , 387 claim 1 , 393 claim 1 , 399 claim 1 , 405 claim 1 , 419 claim 1 , 420 claim 1 , 422 claim 1 , 431 claim 1 , 432 claim 1 , 433 claim 1 , 434 claim 1 , 434 claim 1 , 444 claim 1 , 453 claim 1 , 456 claim 1 , 459 claim 1 , 469 claim 1 , 470 claim 1 , 475 claim 1 , 480 claim 1 , 485 claim 1 , 493 claim 1 , 504 claim 1 , 505 claim 1 , 510 claim 1 , 513 claim 1 , 515 claim 1 , 525 claim 1 , and/or 526 claim 1 , wherein the residue position(s) are numbered with respect to SEQ ID NO: 3.3. The mutant soluble diiron monooxygenase hydroxylase alpha subunit according to claim 2 , wherein the one or more mutations comprises substitutions at one or more of residue position(s): L3P claim 2 , L14P claim 2 , A25S claim 2 , Q26G claim 2 , L32P claim 2 , Q33K claim 2 , Q33R claim 2 , F35L claim 2 , N42R claim 2 , T44A claim 2 , T44S claim 2 , A53G claim 2 , N54K claim 2 , N54L claim 2 , E55A ...

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Номер записи: 69
05-07-2018 дата публикации

CELLULOLYTIC COMPOSITIONS COMPRISING MONOOXYGENASE POLYSACCHARIDE ENZYMES WITH IMPROVED ACTIVITY

Номер: US20180187167A1
Автор: ÁLVAREZ NÚÑEZ Consolación, Bermudez Alcantara Maria de los Angeles, DÍEZ GARCÍA Bruno, GAVALDÁ MARTÍN Sandra, LEDESMA GARCÍA Laura, MARTÍN PÉREZ Lucia, MORENO PÉREZ Antonio Javier, PÉREZ GÓMEZ Dolores, PLATERO GÓMEZ Ana Isabel, RAMOS MARTÍN Juan Luis, REYES SOSA Francisco Manuel, ROCHA MARTÍN Javier, SÁNCHEZ ZAMORANO Laura, VALBUENA CRESPO Noelia, VIÑAS DE LA CRUZ Laura
Принадлежит:

The invention refers to methods and compositions for stabilizing and increasing the activity of enzymatic mixtures comprising GH61 (PMO or polysaccharide monooxigenase) polypeptides used for the degradation of cellulosic material during the saccharification step of biofuel production processes. This improvement is achieved by the addition of a nickel cation to said enzymatic mixtures before and/or during the saccharification step. Thus, the invention provides compositions comprising PMOs, cellulolytic enzymes and a nickel cation, as well as methods for preparing said compositions and methods for producing fermentable sugars and bioproducts, preferably bioethanol, from cellulosic biomass in which said compositions are used. 1. A composition comprising at least one polysaccharide monooxygenase enzyme and a nickel cation.2. The composition according to wherein the nickel cation is present at a concentration of more than 0.0001 mM and less than 50 mM claim 1 , preferably between 0.001 and 20 mM claim 1 , more preferably between 0.001 and 5 mM claim 1 , even more preferably between 0.05 and 5 mM claim 1 , even more preferably between 0.05 and 0.5 mM and even more preferably between 0.075 and 0.125 mM.3. The composition according to any of or wherein the nickel cation is in the form of a salt.4. The composition according to wherein the nickel salt is selected from nickel sulphate claim 3 , nickel chloride claim 3 , nickel nitrate claim 3 , nickel acetate or nickel hydroxide claim 3 , or any combination thereof.5. The composition according to any of to wherein the polysaccharide monooxygenase enzyme is selected from PMO1 claim 3 , PMO2 claim 3 , PMO3 or any combination thereof.6. The composition according to any of to wherein the composition further comprises cellulolytic enzymes.7. The composition according to wherein the cellulolytic enzymes are selected from endoglucanase claim 6 , beta-glucosidase claim 6 , cellobiohydrolase claim 6 , beta-xylosidase claim 6 , ...

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Номер записи: 70
12-07-2018 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20180195053A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity , wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO:136 , and one or more amino acid substitutions at one or more positions corresponding to positions in SEQ ID NO: 136 , selected from the group consisting of 37 , 277 , 278 , 280 , 281 , 326 , 432 , 433 , 435 , and 490.3. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO:136.4. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide is further capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% enantiomeric excess.5. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring ...

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Номер записи: 71
13-07-2017 дата публикации

PREPARATION OF 7-DEHYDROCHOLESTEROL AND/OR THE BIOSYNTHETIC INTERMEDIATES AND/OR SECONDARY PRODUCTS THEREOF IN TRANSGENIC ORGANISMS

Номер: US20170198328A1
Автор: Lang Christine, VEEN Markus
Принадлежит: ORGANOBALANCE GMBH

The present invention relates to a method for preparing 7-dehydrocholesterol and/or the biosynthetic intermediates and/or secondary products thereof by culturing organisms, in particular yeasts. Furthermore, the invention relates to the preparation of the nucleic acid constructs required for preparing the genetically modified organisms and to said genetically modified organisms, in particular yeasts, themselves. 1. A method for preparing 7-dehydrocholesterol and/or the biosynthetic intermediates and/or secondary products thereof comprising the step of culturing organisms which , compared to the wild type , have an increased activity of at least one of the activities selected from the group consisting of Δ8-Δ7-isomerase activity , Δ5-desaturase activity and Δ24-reductase activity.2. The method of claim 1 , wherein the organisms claim 1 , compared to the wild type claim 1 , have an increased activity of at least two of the activities selected from the group consisting of Δ8-Δ7-isomerase activity claim 1 , Δ5-desaturase activity and Δ24-reductase activity.3. The method of claim 1 , wherein the organisms claim 1 , compared to the wild type claim 1 , have an increased Δ8-Δ7-isomerase activity claim 1 , Δ5-desaturase activity and Δ24-reductase activity.4. The method of claim 1 , further comprising the step of increasing the Δ8-Δ7-isomerase activity by increasing claim 1 , compared to the wild type claim 1 , gene expression of a nucleic acid encoding a Δ8-Δ7-isomerase.5. The method of claim 4 , wherein the gene expression is increased by introducing into the organism one or more nucleic acids encoding a Δ8-Δ7-isomerase.6. The method of claim 5 , further comprising the step of introducing the nucleic acids claim 5 , which encode proteins comprising the amino acid sequence SEQ. ID. NO. 2 or a sequence derived from this sequence by substitution claim 5 , insertion or deletion of amino acids claim 5 , which is at least 30% identical at the amino acid level with the sequence ...

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Номер записи: 72
27-07-2017 дата публикации

BIOSYNTHETIC PRODUCTION OF ACETAMINOPHEN, P-AMINOPHENOL, AND P-AMINOBENZOIC ACID

Номер: US20170211104A1
Автор: ANDERSON JOHN CHRISTOPHER, Daly Mark T., Poon Patrick, Revak Timothy, Srivastava Saurabh
Принадлежит:

The present disclosure provides compositions and methods for the biosynthetic production of acetaminophen, p-aminophenol, and p-aminobenzoic acid and the purification of biologically derived acetaminophen. 1. A non-naturally occurring microbial organism comprising at least three exogenous genes encoding acetaminophen pathway enzymes expressed in a sufficient amount to produce acetaminophen , wherein said acetaminophen pathway comprises (i) an enzyme that converts chorismic acid to p-aminobenzoic acid (ii) an enzyme that converts p-aminobenzoic acid to p-aminophenol and (iii) an enzyme that converts p-aminophenol to acetaminophen.2. The non-naturally occurring microbial organism of wherein said enzyme that converts chorismic acid to p-aminobenzoic acid is a two protein complex comprising ADC synthase and aminodeoxychorismate lyase; wherein said enzyme that converts p-aminobenzoic acid to p-aminophenol is 4-aminobenzoate 1-monoygenase and wherein said enzyme that converts p-aminophenol to acetaminophen is N-hydroxyarylamine O-acetyltransferase.3. The non-naturally occurring microbial organism of wherein said N-hydroxyarylamine O-acetyltransferase comprises SEQ ID NO: 4 or the active domain thereof.4. The non-naturally occurring microbial organism of wherein said enzyme that converts chorismic acid to p-aminobenzoic acid is a two protein complex comprising ADC synthase and aminodeoxychorismate lyase; wherein said enzyme that converts p-aminobenzoic acid to p-aminophenol is 4-aminobenzoate 1-monoygenase and wherein said enzyme that converts p-aminophenol to acetaminophen is arylamine N-acetyltransferase.5. The non-naturally occurring microbial organism of wherein said arylamine N-acetyltransferase comprises SEQ ID NO: 5 or the active domain thereof.69.-. (canceled)10. The non-naturally occurring microbial organism of wherein said ADC synthase comprises SEQ ID NO: 1 or the active domain thereof and said aminodeoxychorismate lyase comprises SEQ ID NO: 2 or the active ...

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Номер записи: 73
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: 74
18-07-2019 дата публикации

MODULATION OF FLUID INTAKE

Номер: US20190216948A1
Автор: Augustine Vineet, OKA Yuki
Принадлежит:

In some embodiments, methods of stimulating fluid intake in a subject in need thereof are described. The methods can comprise stimulating a nitric oxide synthase (nNOS)-positive neuron of the median preoptic nucleus (MnPO). In some embodiments, methods of inhibiting fluid intake in a subject in need thereof are described. The methods can comprise inhibiting stimulation of an nNOS-positive neuron of the MnPO. 1. A method of stimulating fluid intake in a subject in need thereof , the method comprising , in a first nitric oxide synthase (nNOS)-positive neuron of the median preoptic nucleus (MnPO) comprising a polarized cell membrane , stimulating depolarization of the cell membrane ,thereby stimulating the first nNOS-positive neuron, thereby stimulating fluid intake in the subject.2. The method of claim 1 , wherein stimulating depolarization of the cell membrane comprises at least one of a net influx of cations into a cytosol of the first nNOS-positive neuron claim 1 , a net efflux of anions from the cytosol of the first nNOS-positive neuron claim 1 , and/or stimulating a second nNOS-positive neuron of the subfornical organ (SFO).3. The method of claim 1 , wherein stimulating depolarization of the cell membrane of the first nNOS-positive neuron comprises:administering a nucleic acid encoding a stimulatory conditional ion modulator to the subject, the stimulatory conditional ion modulator configured to induce a net influx of cations into the cytosol of the first nNOS-positive neuron and/or a net efflux of anions from the cytosol of the first nNOS-positive neuron in response to a stimulus or agonist,whereby the nucleic acid is under the control of a promoter that induces transcription of the nucleic acid in the first nNOS-positive neuron, whereby the conditional ion modulator is expressed in the first nNOS-positive neuron; andapplying an agonist or stimulus to the first nNOS-positive neuron of the subject, causing the conditional ion modulator to induce the net influx of ...

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Номер записи: 75
19-08-2021 дата публикации

HETEROLOGOUS EXPRESSION OF SHORT-CHAIN MONOOXYGENASES IN MICROORGANISMS

Номер: US20210254026A1
Автор: CLARKE Elizabeth Jane, Greenfield Derek Lorin, Helman Noah Charles, Jones Stephanie Rhianon, Zhu Baolong
Принадлежит:

Methods and compositions for the oxidation of short alkanes by engineered microorganisms expressing enzymes are described, along with methods of use. 1. A synthetic polynucleotide for a soluble diiron monooxygenase enzyme which can be expressed in a microorganism of interest , comprising at least one monooxygenase coding region encoding a diiron monooxygenase enzyme , the at least one monooxygenase coding region linked to at least one promoter which will function in the microorganism of interest.2. The synthetic polynucleotide of claim 1 , further comprising at least one protein folding chaperone coding region encoding at least one protein folding chaperone claim 1 , the at least one protein chaperone coding region linked to at least one promoter which will function in the microorganism of interest.3. The synthetic polynucleotide of claim 1 , further comprising at least one mutation claim 1 , wherein the at least one mutation increases specificity for a monooxygenase substrate and/or increases production of a chemical.4. The synthetic polynucleotide of claim 2 , further comprising at least one mutation claim 2 , wherein the at least one mutation increases specificity for a monooxygenase substrate and/or increases production of a chemical.5. The synthetic polynucleotide of or claim 2 , wherein the soluble diiron monooxygenase enzyme is at least 60% identical to SEQ ID NO: 7 or SEQ ID NO: 9 or SEQ ID NO: 11 or SEQ ID NO: 13 or SEQ ID NO: 58 or SEQ ID NO: 60 or SEQ ID NO: 87 or SEQ ID NO: 89 or SEQ ID NO: 91 or SEQ ID NO: 93 or SEQ ID NO: 95 or SEQ ID NO: 97 or SEQ ID NO: 99 or SEQ ID NO: 101 or SEQ ID NO: 103 or SEQ ID NO: 105 or SEQ ID NO: 107 or SEQ ID NO: 109 or SEQ ID NO: 111 or SEQ ID NO: 113 or SEQ ID NO: 115 or SEQ ID NO: 117 or SEQ ID NO: 143 or SEQ ID NO: 145 or SEQ ID NO: 147 or SEQ ID NO: 149 or SEQ ID NO: 151 or SEQ ID NO: 153.6. The synthetic polynucleotide of or claim 2 , wherein the at least one mutation is a E to N at position 240 in SEQ ID NO: 10.7. A ...

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Номер записи: 76
19-08-2021 дата публикации

Herbicide tolerant plants expressing a cyanobacterial plastoquinone biosynthetic pathway

Номер: US20210254091A1
Автор: Manuel Dubald
Принадлежит: BASF Agricultural Solutions Seed US LLC

A method for conferring tolerance to a 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicide in a plant by expressing one or more polypeptide components of an exogenous plastoquinone-9 pathway in the plant. Nucleic acids, vectors, transgenic cells and transgenic plants useful in such a method are also disclosed.

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Номер записи: 77
09-07-2020 дата публикации

REGULATING PROTEIN SPONTANEOUS RUPTURES

Номер: US20200216824A1
Автор: Lu H. Peter
Принадлежит: BOWLING GREEN STATE UNIVERSITY

Protein rupture under compressive forces can be regulated by cations. More specifically, pico-Newton forces can cause rupture of protein molecules, as shown in examples with calmodulin (CaM) and tau proteins, among others. However, rupture does not occur in the presence of various concentrations of cation(s), thus elucidating new targets for disease therapy and providing therapies for neurodegenerative diseases or other conditions involving protein misfolding, dysfunction, or aggregation. 1. A method for preventing or reducing rupture of a protein from a compressive force , the method comprising exposing the protein to a concentration of a cation effective to prevent or reduce rupture of the protein from the compressive force.2. The method of claim 1 , wherein the protein is present in a human cell.3. The method of claim 1 , wherein the protein is a globular protein.4. The method of claim 1 , wherein the protein is selected from the group consisting of tau claim 1 , HPPK claim 1 , nitric oxide synthase (NOS) claim 1 , and calmodulin (CaM).5. The method of claim 1 , wherein the cation is selected from the group consisting of metallic cations and organic cations.6. The method of claim 1 , wherein the cation comprises Ca.7. The method of claim 1 , wherein the cation comprises Mg.8. The method of claim 1 , wherein the cation comprises Zn.9. The method of claim 1 , wherein the compressive force is at least about 12 pN.10. The method of claim 1 , wherein the protein is in an aqueous solution.11. The method of claim 10 , wherein the aqueous solution is present in a living cell of a subject selected from the group consisting of human claim 10 , mammal claim 10 , and other animal.12. The method of claim 11 , wherein the subject has a neurodegenerative disease or other condition involving protein misfolding claim 11 , dysfunction claim 11 , or aggregation.13. The method of claim 1 , wherein the concentration of the cation is a micromolar concentration.14. The method of claim ...

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Номер записи: 78
09-07-2020 дата публикации

SYNTHETIC METABOLIC FUNNELING FOR BIOCHEMICAL PRODUCTION

Номер: US20200216865A1
Автор: NIELSEN David, Thompson Brian
Принадлежит: Arizona Board of Regents on behalf of Arizona State University

Certain embodiments provide a method for preparing a biochemical product (e.g., phenol, catechol, or muconic acid, or a salt thereof). For example, such methods include contacting a recombinant host having two or more recombinant pathways with a fermentable carbon source and growing the recombinant cell for a time sufficient to synthesize the product. In certain embodiments, each recombinant pathway: 1) is capable of producing the same final biochemical product; 2) comprises at least one gene encoding a polypeptide; 3) is derived from a different endogenous metabolite as its immediate precursor; and 4) converges to the same final product or the same intermediate metabolite. 1. A method for preparing a biochemical product , the method comprising: a) each pathway is capable of producing the same final biochemical product;', 'b) each pathway comprises at least one gene encoding a polypeptide;', 'c) each pathway is derived from a different endogenous metabolite as its immediate precursor; and', 'd) each pathway converges to the same final product or the same intermediate metabolite; and, 'i) contacting a recombinant host cell with a fermentable carbon source, wherein the recombinant host comprises two or more recombinant pathways, and whereinii) growing said recombinant cell for a time sufficient to synthesize the product.2. The method of claim 1 , wherein each pathway converges to the same final product.3. The method of claim 1 , wherein each pathway converges to the same intermediate metabolite claim 1 , further comprising each pathway continuing to the same final product.4. A method for preparing a biochemical product claim 1 , the method comprising: a) each pathway is capable of producing the same final biochemical product;', 'b) each pathway comprises at least one gene encoding a polypeptide;', 'c) each pathway is derived from the same endogenous metabolite as its immediate precursor;', 'd) each pathway proceeds via different intermediate metabolites;', 'e) each ...

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Номер записи: 79
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: 80
16-07-2020 дата публикации

METHODS FOR PRODUCTION OF OXYGENATED TERPENES

Номер: US20200224224A1
Автор: Ghosh Souvik, KUMARAN Ajikumar Parayil, LI Liwei, Lim Chin Giaw, Marshall-Hill Geoff, Pirie Christopher, Preininger Martin, Qualley Anthony
Принадлежит:

The present disclosure relates to methods for producing oxygenated terpenoids, and preparation of compositions and formulations thereof. Polynucleotides, derivative enzymes, and host cells for use in such methods are also provided. 1182-. (canceled)183. A method of making a formulation comprising nootkatone , α-nootkatol , and β-nootkatol , the method comprising:{'i': 'Stevia rebaudiana', 'contacting valencene with Kaurene Oxidase (SrKO) or an SrKO derivative having valencene oxidizing activity in an in vitro or in vivo system to produce an oxygenated product comprising nootkatone, α-nootkatol, and β-nootkatol.'}184. The method of claim 183 , further comprising processing the oxygenated product by fractional distillation to yield two or more fractions of the oxygenated product.185. The method of claim 184 , further comprising blending the two or more fractions of the oxygenated product.186. The method of claim 183 , further comprising adding an unoxygenated product to the oxygenated product claim 183 , one or more fractions of the oxygenated product claim 183 , or the formulation.187. The method of claim 183 , wherein the nootkatone is present in the formulation in an amount ranging from about 55% to about 60% (w/w) claim 183 , the a-nootkatol is present in the formulation in an amount ranging from about 20% to about 25% (w/w) claim 183 , and the β-nootkatol is present in the formulation an amount ranging from about 5% to about 10% (w/w).188. The method of claim 186 , wherein the unoxygenated product comprises valencene.189. The method of claim 188 , wherein the valencene is present in the formulation in an amount ranging from about 5% to about 10% (w/w). This application is a divisional of U.S. patent application Ser. No. 15/505,022, filed Feb. 17, 2017, which is a United States National Stage Application of International Application No. PCT/US/2015/046421, filed Aug. 21, 2015, which claims the benefit of, and priority to, United States Provisional Application No. ...

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Номер записи: 81
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: 82
26-08-2021 дата публикации

IMMOBILIZED ENZYMES AND MICROSOMES ON MAGNETIC SCAFFOLDS

Номер: US20210263001A1
Автор: Chun Matthew Steven, Corgie Stephane Cedric, Hoepker Alexander Chris, Rivera Katia Argelia Rodriguez, Wong Braedon Carter
Принадлежит: ZYMtronix Catalytic Systems, Inc.

The present invention provides devices and methods for producing metabolites used to measure the toxicity of chemical compounds. They incorporate enzymatic microsomes and magnetic nanoparticles that magnetically entrap enzymes. These enzyme systems catalyze chemicals to yield measurable metabolic products. The microsomes and the magnetic nanoparticles containing enzymes are associated with macroporous scaffolds and non-reactive components that facilitate the enzyme reactions. 1. A device , comprising microsomes and self-assembled mesoporous aggregates of magnetic nanoparticles , wherein a first enzyme requiring a diffusible cofactor having a first enzymatic activity is contained within said microsomes , wherein a second enzyme comprising a cofactor regeneration activity is magnetically-entrapped within said mesopores ,wherein said cofactor is utilized in said first enzymatic activity;wherein said first and second enzymes function by converting a diffusible substrate into a diffusible product;wherein said magnetic nanoparticles are magnetically associated with a macroporous scaffold;wherein said microsomes are associated with said macroporous scaffold; andwherein said macroporous scaffold comprising said magnetic nanoparticles is associated with a non-reactive portion operable for placing said macroporous scaffold into or removing it from a reaction solution.2. The device of claim 1 , further comprising a functional portion for stably maintaining said microsomes claim 1 , said magnetic nanoparticles claim 1 , said first enzyme claim 1 , said second enzyme claim 1 , and said macroporous scaffold.3. The device of claim 2 , wherein said functional portion comprises a buffer.4. The device of claim 2 , wherein said functional portion comprises a substrate for said second enzyme.5. The device of claim 2 , wherein said functional portion comprises said cofactor.6. The device of claim 2 , wherein said functional portion is magnetic.7. The device of any one of - claim 2 , ...

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Номер записи: 83
25-08-2016 дата публикации

RECOMBINANT ACETOGENIC BACTERIUM FOR THE CONVERSION OF METHANE TO PRODUCTS

Номер: US20160244785A1
Автор: Dürre Peter, Koepke Michael
Принадлежит:

The invention provides a recombinant, acetogenic bacterium that consumes a substrate comprising CHand converts at least a portion of the CHto a product. In particular, the bacterium of may comprise one or more of exogenous methane monooxygenase (MMO), exogenous nitrite reductase (NIR), and exogenous nitric oxide dismutase (NOD). The invention further provides a method for producing a product comprising providing a substrate comprising CHto a culture comprising a recombinant, acetogenic bacterium, whereby the bacterium converts at least a portion of the CHto a product. 1. A recombinant , acetogenic bacterium that consumes a substrate comprising CHand converts at least a portion of the CHto a product.2. The bacterium of claim 1 , wherein the bacterium comprises one or more of exogenous methane monooxygenase (MMO) claim 1 , exogenous nitrite reductase (NIR) claim 1 , and exogenous nitric oxide dismutase (NOD).3. The bacterium of claim 2 , wherein the methane monooxygenase claim 2 , nitrite reductase claim 2 , or nitric oxide dismutase are derived from Methylomirabilis oxyfera.4. The bacterium of claim 2 , wherein the bacterium further comprises exogenous methanol methyltransferase.5ClostridiumAcetobacterium.. The bacterium of claim 1 , wherein the bacterium is a member of genus or6Clostridium autoethanogenum, Clostridium ljungdahlii, Clostridium ragsdalei,Acetobacterium woodii.. The bacterium of claim 1 , wherein the bacterium is derived from or7. The bacterium of claim 1 , wherein the substrate further comprises one or more of CO claim 1 , CO claim 1 , and H.8. The bacterium of claim 1 , wherein the substrate further comprises one or more of NO and NO.9. The bacterium of claim 1 , wherein CHis the sole carbon source for the bacterium.10. The bacterium of claim 1 , wherein the product comprises one or more of ethanol claim 1 , acetate claim 1 , butanol claim 1 , butyrate claim 1 , 2 claim 1 ,3-butanediol claim 1 , lactate claim 1 , butene claim 1 , butadiene claim 1 , ...

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Номер записи: 84
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: 85
07-09-2017 дата публикации

Nucleic acid constructs including a txnip promoter for the treatment of disease

Номер: US20170252376A1
Автор: Lalit Singh Pukhrambam
Принадлежит: WAYNE STATE UNIVERSITY

Nucleic acids for the treatment of diseases are described. The nucleic acids include a thioredoxin-interacting protein (TXNIP) promoter and a gene that encodes a therapeutic protein or an interfering nucleic acid sequence (e.g., interfering RNA (iRNA sequence)).

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Номер записи: 86
15-09-2016 дата публикации

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

Номер: US20160264945A1
Автор: ALVIZO Oscar, Ang Ee Lui, Behrouzian Behnaz, Clay Michael D., Collier Steven J., Eberhard Ellen D., Fan Fu, Smith Derek J., Song Shiwei, Widegren Magnus, Wilson Robert, Xu Junye, ZHU Jun
Принадлежит:

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO:2 and one or more amino acid differences relative to SEQ ID NO: 2 at positions selected from 277 , 281 , 326 , 433 , or 435.2. The non-naturally occurring polypeptide sequence of claim 1 , wherein said polypeptide is capable of converting the acid substrate compound (1b) to compound (2b) (R-enantiomer) or its opposite enantiomer compound (S-enantiomer) with at least 2-fold improved activity relative to the wild-type polypeptide of SEQ ID NO: 2.3. The non-naturally occurring polypeptide of claim 1 , wherein said polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 selected from: X246A claim 1 , E claim 1 , G claim 1 , I claim 1 , L claim 1 , P claim 1 , S claim 1 , T claim 1 , and V.4. The non-naturally occurring polypeptide of claim 1 , wherein said polypeptide is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% ee.5. The non-naturally occurring polypeptide of claim 1 , wherein said polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 selected from: X143G claim 1 , X278G claim 1 , X326R claim 1 , and X490L.6. The non-naturally occurring polypeptide of claim 1 , wherein said ...

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Номер записи: 87
15-09-2016 дата публикации

ENGINEERED RECOMBINANT ENZYMES FOR METHANE OXIDATION

Номер: US20160265003A1
Автор: Dousis Athanasios D., Kostecki Jan S., Lawton Thomas J., Lee Fredrick, Nisthal Alex, Olafson Barry, Privett Heidi K., Rosenzweig Amy C
Принадлежит:

Provided herein are soluble engineered polypeptides for oxidizing hydrocarbons, and methods of use, manufacture, and design thereof. In particular, soluble, polypeptides capable of oxidizing methane to methanol (e.g., hydroxylation) are provided. 1. A soluble polypeptide comprising a core sequence having at least 60% but less than 100% sequence identity with SEQ ID NO: 23 , wherein the soluble polypeptide is capable of converting an alkane into an alkanol , and wherein the core sequence is C-terminally- and/or N-terminally-flanked by one or more soluble peptide segments.2. The soluble polypeptide of claim 1 , wherein the polypeptide is capable of converting methane into methanol.3. The soluble polypeptide of claim 1 , wherein the polypeptide carries out the hydroxylation of the alkane.4. The soluble polypeptide of claim 1 , wherein the polypeptide comprises a copper oxidase with an engineered active site.6. The soluble polypeptide of claim 1 , wherein the polypeptide is expressed in an industrially-relevant host organism and at a commercially-relevant scale.7. The soluble polypeptide of claim 1 , wherein the one or more soluble peptide segments are selected from peptides having at least 60% sequence identity with one or SEQ ID NOS: 16 claim 1 , 17 claim 1 , 18 claim 1 , 21 claim 1 , and 22.9. The soluble polypeptide of claim 1 , having at least 60% sequence identity with spmoB7 (SEQ ID NO: 13) claim 1 , sumo-spmoB7 (SEQ ID NO: 14) claim 1 , and/or sumo-spmoB7 8pt3 (SEQ ID NO: 13) claim 1 , but less than 100% sequence identify from a naturally-occurring sequence.10. A method of converting an alkane into an alkanol comprising exposing the alkane to the soluble polypeptide .11. A cell expressing the soluble polypeptide of .12. A method of producing a biofuel comprising exposing an alkane to a soluble polypeptide of .13. A system comprising a soluble polypeptide of attached to a fixed support.14. The system of claim 13 , wherein the fixed support is selected from the ...

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Номер записи: 88
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: 89
15-08-2019 дата публикации

Method for Producing Objective Substance

Номер: US20190249206A1
Автор: Asari Sayaka, FUKUI Keita, Mijts Benjamin, Roche Christine, Toyazaki Miku
Принадлежит: AJINOMOTO CO., INC.

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 enolase is reduced. 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 enolase is reduced as compared with a non-modified microorganism, andwherein 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 the culture broth claim 5 , cells present in a processed product of ...

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Номер записи: 90
14-10-2021 дата публикации

METHODS OF TREATING AN AUTOIMMUNE DISEASE

Номер: US20210315967A1
Автор: Gommerman Jennifer, Rojas Olga
Принадлежит:

The present disclosure provides methods of treating an autoimmune disease by administering at least a B-cell Activating Factor (BAFF) polypeptide to a subject in need thereof. 1. A method of treating an autoimmune disease in a subject , the method comprising administering an effective amount of one or more of:a B-cell Activating Factor (BAFF) polypeptide;a BAFF polypeptide and an agent that promotes survival and/or migration of gut-derived commensal-reactive B cells to the central nervous system of the subject; ora BAFF polypeptide and a gut commensal that increases IgA levels to the subject.2. The method of claim 1 , wherein the autoimmune disease is a non-systemic organ-specific autoimmune disease.3. The method of or claim 1 , wherein the autoimmune disease is multiple sclerosis.4. The method of any one of - claim 1 , wherein the commensal-reactive B cells are IgA+ plasmablasts and/or plasma cells.5. The method of any one of - claim 1 , wherein the commensal-reactive B cells express IL-10 and/or iNOS.6. The method of any one of - claim 1 , wherein the agent that promotes survival and/or migration of gut-derived commensal-reactive B cells is a cytokine or a chemokine.7. The method of any one of - claim 1 , wherein the agent that promotes survival and/or migration of gut-derived commensal-reactive B cells is IL-10 and/or iNOS.8. The method of any one of - claim 1 , wherein the gut commensal is a commensal microbe.9. The method of any one of - claim 1 , wherein the administering an effective amount of a gut commensal comprises oral or rectal administration of a microbe or community of microbes.10Tritrichomonas musculisTritrichomonas musculis.. The method of any one of - claim 1 , wherein the gut commensal comprises or a gut microbial community that has been modified by carriage of11. A method of reducing inflammation in a subject claim 1 , the method comprising administering an effective amount of one or more of:a B-cell Activating Factor (BAFF) polypeptide;a BAFF ...

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Номер записи: 91
14-09-2017 дата публикации

METHOD FOR BIOSYNTHESIS OF ACETAMINOPHEN

Номер: US20170260555A1
Автор: ANDERSON JOHN CHRISTOPHER, Bodik Rastislav, HSIAU Timothy, Kotker Jonathan Prakash Inocencio, Ruan Paul, Seshia Sanjit A., Srivastava Saurabh
Принадлежит:

The present disclosure provides methods for biosynthesis of acetaminophen. The present disclosure provides host cells genetically modified to provide for production of acetaminophen. The present disclosure provides a recombinant host cell that is genetically modified with one or more heterologous nucleic acids comprising nucleotide sequences encoding 4-aminobenzoate hydroxylase (4ABH) and N-hydroxyarylamine O-acetyltransferase (NhoA). The present disclosure provides a recombinant prokaryotic host cell that is genetically modified with one or more heterologous nucleic acids comprising nucleotide sequences encoding 44ABH and NhoA. 1. A recombinant prokaryotic host cell that is genetically modified with one or more heterologous nucleic acids comprising nucleotide sequences encoding 4-aminobenzoate hydroxylase (4ABH) and N-hydroxyarylamine O-acetyltransferase (NhoA).2. The recombinant prokaryotic host cell of claim 1 , wherein the host cell comprises one or more endogenous nucleic acids comprises nucleotide sequences encoding polypeptides comprising amino acid sequences having at least 75% amino acid sequence identity to the amino acid sequences set forth in SEQ ID NOs:3 claim 1 , 4 claim 1 , and 5.3. The recombinant prokaryotic host cell of claim 1 , wherein the host cell is genetically modified with one or more heterologous nucleic acids comprising nucleotide sequences encoding polypeptides comprising amino acid sequences having at least 75% amino acid sequence identity to the amino acid sequences set forth in SEQ ID NOs:3 claim 1 , 4 claim 1 , and 5.4. The recombinant prokaryotic host cell of claim 3 , wherein the nucleotide sequences encoding polypeptides comprising amino acid sequences having at least 75% amino acid sequence identity to the amino acid sequences set forth in SEQ ID NOs:3 claim 3 , 4 claim 3 , and 5 are present on a single expression vector.5. The recombinant prokaryotic host cell of or claim 3 , wherein the nucleotide sequences encoding polypeptides ...

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Номер записи: 92
20-09-2018 дата публикации

RNA PREPARATIONS COMPRISING PURIFIED MODIFIED RNA FOR REPROGRAMMING CELLS

Номер: US20180265848A1
Автор: Dahl Gary, Jendrisak Jerome, Kariko Katalin, Meis Judith, Person Anthony, Weissman Drew
Принадлежит:

The present invention provides compositions and methods for reprogramming somatic cells using purified RNA preparations comprising single-strand mRNA encoding an iPS cell induction factor. The purified RNA preparations are preferably substantially free of RNA contaminant molecules that: i) would activate an immune response in the somatic cells, ii) would decrease expression of the single-stranded mRNA in the somatic cells, and/or iii) active RNA sensors in the somatic cells. In certain embodiments, the purified RNA preparations are substantially free of partial mRNAs, double-stranded RNAs, un-capped RNA molecules, and/or single-stranded run-on mRNAs. 1. A method for reprogramming a human or animal cell that exhibits a first differentiated state or phenotype to a cell that exhibits a second differentiated state or phenotype comprising: introducing into the cell that exhibits a first differentiated state a purified RNA preparation comprising modified mRNA molecules that encode at least one reprogramming factor and culturing the cell under conditions wherein the cell exhibits a second differentiated state.2. The method of claim 1 , wherein the modified mRNA molecules contain at least one modified nucleoside selected from the group consisting of: pseudouridine (ψ) claim 1 , 5-methylcytosine (mC) claim 1 , 5-methyluridine (mU) claim 1 , 2′-O-methyluridine (Um or mU) claim 1 , 2-thiouridine (sU) claim 1 , and N-methyladenosine (mA)) in place of at least a portion of the corresponding unmodified canonical nucleoside.3. The method of claim 2 , wherein the at least one modified nucleoside is present in place of substantially all of the corresponding unmodified canonical nucleoside.4. The method of claim 1 , wherein said purified RNA preparation:i) comprises at least one single-stranded mRNA that encodes an iPS cell induction factor, wherein substantially all of said first single-stranded mRNAs comprise at least one pseudouridine residue and/or at least one 5-methylcytidine ...

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Номер записи: 93
11-11-2021 дата публикации

KAURENOIC ACID HYDROXYLASES

Номер: US20210348136A1
Автор: BOER Viktor Marius, VAN LEEUWEN Johannes Gustaaf Ernst, ZWARTJENS Priscilla
Принадлежит:

The present invention relates a polypeptide having kaurenoic acid 13-hydroxylase activity, which 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 or SEQ ID NO: 3, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 136, 248, 336 or 403, said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside. 1. A polypeptide having kaurenoic acid 13-hydroxylase activity , which 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 or SEQ ID NO: 3 , comprises at least one substitution of an amino acid corresponding to any of amino acids at positions136, 248, 336 or 403said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity.2. The polypeptide according to wherein the positions in the polypeptide having kaurenoic acid 13-hydroxylase activity corresponding to any amino acids at position 136 claim 1 , 248 claim 1 , 336 or 403 claim 1 , said position being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 claim 1 , are identified by aligning the amino acid sequence of the polypeptide with kaurenoic acid 13-hydroxylase activity with the amino acid sequence set out in SEQ ID NO: 1 or SEQ ID NO: 3 using the EMBOSS Needle alignment method claim 1 , using EBLOSUM62 as a substitution matrix claim 1 , with a gap-open penalty of 10 and a gap extension penalty of 0.5.3. The polypeptide according ...

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Номер записи: 94
29-08-2019 дата публикации

Compositions And Methods For Production Of Salidroside, Icariside D2, And Precursors Of Salidroside And Icariside D2

Номер: US20190264221A1
Автор: Torrens-Spence Michael, Weng Jing-Ke
Принадлежит:

Transgenic host cells, vectors useful for making transgenic host cells, and kits useful for making transgenic host cells are described. Also described are transgenic plants. In some embodiments, transgenic host cells express a 4-hydroxyphenylacetaldehyde synthase (4HPAAS). In some embodiments, transgenic host cells express a tyrosol:UDP-glucose 8-O-glucosyltransferase (T8GT). The transgenic host cells are useful for biosynthesis of one or more of salidroside, icariside D2, tyrosol, and 4-hydroxypenylacetaldehyde. 179-. (canceled)80. A host cell comprising a transgene encoding a 4-hydroxyphenylacetaldehyde synthase (4HPAAS) , wherein the 4HPAAS comprises an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 2 , and wherein the amino acid sequence of the 4HPAAS comprises:a) an amino acid residue selected from the group consisting of F, L, I, M and V at a position corresponding to the F residue at position 343 in SEQ ID NO: 2;b) an amino acid residue selected from the group consisting of N and D at a position corresponding to the H residue at position 198 in SEQ ID NO: 2; orc) a combination thereof.81. The host cell of claim 80 , wherein the amino acid sequence of the 4HPAAS comprises an amino acid residue selected from the group consisting of F claim 80 , L claim 80 , I claim 80 , M and V at a position corresponding to the F residue at position 343 in SEQ ID NO: 2.82. The host cell of claim 80 , wherein the amino acid sequence of the 4HPAAS comprises an amino acid residue selected from the group consisting of N and D at a position corresponding to the H residue at position 198 in SEQ ID NO: 2.83. The host cell of claim 80 , wherein the amino acid sequence of the 4HPAAS comprises an amino acid residue selected from the group consisting of F claim 80 , L claim 80 , I claim 80 , M and V at a position corresponding to the F residue at position 343 in SEQ ID NO: 2 claim 80 , and an amino acid residue selected from the group consisting of N ...

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Номер записи: 95
05-09-2019 дата публикации

MICROBIAL PRODUCTION OF STEVIOL GLYCOSIDES

Номер: US20190269157A1
Автор: Donald Jason, Gupta Swati, KUMARAN Ajikumar Parayil, LI Liwei, LIM Ryan, PATEL Krishna, PHILIPPE Ryan
Принадлежит:

The invention provides methods for making steviol glycosides, including RebM and glycosylation products that are minor products in stevia leaves, and provides enzymes, encoding polynucleotides, and host cells for use in these methods. The invention provides engineered enzymes and engineered host cells for producing steviol glycosylation products, such as RebM, at high purity and/or yield. The invention further provides methods of making products containing steviol glycosides, such as RebM, including food products, beverages, oral care products, sweeteners, and flavoring products. 180.-. (canceled)81. A method for producing Rebaudioside M (RebM) , comprising:{'i': 'E. coli', 'providing an host cell expressing recombinant uridine diphosphate-dependent glycosyl transferase (UGT) enzymes catalyzing the following glycosyltransferase reactions on steviol or steviol glycoside substrate: 13-O glycosylation, 19-O glycosylation, 1-2′ glycosylation at C-19 and C-13, and 1-3′ glycosylation at C-19 and C-13;'}{'i': 'E. coli', 'culturing the host cell in a culture medium under conditions suitable for producing RebM, wherein the host cell transports RebM into the culture medium; and'}recovering RebM from the culture medium.82. The method of claim 81 , wherein RebM is produced in the culture medium at a concentration of at least 100 mg/L.83. The method of claim 81 , wherein RebM is produced in the culture medium at a concentration of at least 1 g/L.84. The method of claim 81 , wherein RebM is produced in the culture medium at a concentration of at least 10 g/L.85. The method of claim 81 , wherein the UGT enzymes comprise one or more of:(a) a modified UGT enzyme having an increase in 1-2′ glycosylating activity at C19 of Rebaudioside A (RebA) as compared to its parent UGT enzyme, without substantial loss of 1-2′ glycosylating activity at C13 of steviolmonoside; and(b) a modified UGT enzyme having an increase in 1-3′ glycosylating activity at C19 of Rebaudioside D (RebD) as compared ...

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Номер записи: 96
03-09-2020 дата публикации

COMPOSITIONS AND METHODS FOR TREATMENT OF SEPSIS-RELATED DISORDERS

Номер: US20200276223A1
Автор: Mansell John
Принадлежит:

A method comprising administering, to a subject in need thereof, an effective amount of a nucleotide effective to disrupt one or more pathways leading to sepsis. The nucleotide may be a nitric oxide disruptor effective to decrease the expression of inducible nitric oxide synthase. The nitric oxide disrupter may comprise a polynucleotide strand exhibiting at least 70% sequence identity to one of Sequence ID No.1 through Sequence ID No. 47. Additionally or alternatively, the nucleotide may be an α disintegrin and metalloproteinase (ADAM) enzyme inhibitor effective to decrease the expression of ADAM enzyme. The ADAM enzyme inhibitor may comprise a polynucleotide strand exhibiting at least 70% sequence identity to one of Sequence ID No. 48 through Sequence ID No. 56. 1. A method comprising administering an effective amount of a nucleotide effective to disrupt one or more pathways leading to sepsis to a subject in need thereof.2. The method of claim 1 , wherein the nucleotide is a nitric oxide disruptor claim 1 , wherein the nitric oxide disrupter is effective to decrease the expression of inducible nitric oxide synthase by at least 50%.3. The method of claim 1 , wherein the nucleotide is a nitric oxide disruptor claim 1 , wherein the nitric oxide disrupter is effective to decrease the expression of inducible nitric oxide synthase by at least 75%.4. The method of claim 2 , wherein the nitric oxide disrupter comprises a polynucleotide strand exhibiting at least 70% sequence identity to one of Sequence ID No.1 through Sequence ID No. 47.5. The method of claim 2 , wherein the nitric oxide disrupter comprises a polynucleotide strand exhibiting at least 80% sequence identity to one of Sequence ID No.1 through Sequence ID No. 47.6. The method of claim 2 , wherein the nitric oxide disrupter comprises a polynucleotide strand exhibiting at least 90% sequence identity to one of Sequence ID No.1 through Sequence ID No. 47.7. The method of claim 2 , wherein the nitric oxide ...

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Номер записи: 97
03-09-2020 дата публикации

SYNTHETIC METHANOTROPHIC AND METHYLOTROPHIC MICROORGANISMS

Номер: US20200277636A1
Автор: CLARKE Elizabeth, GREENFIELD Derek, HELMAN Noah
Принадлежит:

Provided herein are non-naturally occurring microbial organisms comprising a methane-oxidizing metabolic pathway. The invention additionally comprises non-naturally occurring microbial organisms comprising pathways for the production of chemicals. The invention additionally provides methods for using said organisms for the production of chemicals. 1. A synthetic microorganism , wherein said synthetic microorganism comprises a natural methanol-consuming microorganism and one or more genetic modifications that improve the production of a chemical.2. The synthetic microorganism of claim 1 , wherein said chemical is selected from the group consisting of dicarboxylic acid claim 1 , malic acid claim 1 , fumaric acid claim 1 , succinic acid claim 1 , malic acid salt claim 1 , fumaric acid salt claim 1 , succinic acid salt claim 1 , L-malic acid claim 1 , D-malic acid claim 1 , maleic acid claim 1 , lactic acid claim 1 , adipic acid claim 1 , 1 claim 1 ,3-propanediol claim 1 , 2 claim 1 ,3-butanediol claim 1 , 1 claim 1 ,4-butanediol claim 1 , butadiene claim 1 , fatty acid derivatives claim 1 , fatty alcohols claim 1 , fatty acids claim 1 , fatty acid esters claim 1 , fatty acid methyl esters claim 1 , fatty acid ethyl esters claim 1 , branched fatty acids claim 1 , branched fatty acid derivatives claim 1 , omega-3 fatty acids claim 1 , isoprenoids claim 1 , isoprene claim 1 , farnesene claim 1 , farnesane claim 1 , squalene claim 1 , squalane claim 1 , carotenoids claim 1 , amino acids claim 1 , alanine claim 1 , arginine claim 1 , asparagine claim 1 , aspartic acid claim 1 , cysteine claim 1 , glutamic acid claim 1 , monosodium glutamate claim 1 , glutamine claim 1 , glycine claim 1 , histidine claim 1 , isoleucine claim 1 , leucine claim 1 , lysine claim 1 , methionine claim 1 , phenylalanine claim 1 , threonine claim 1 , tryptophan claim 1 , valine claim 1 , ornithine claim 1 , proline claim 1 , selenocysteine claim 1 , serine claim 1 , tyrosine claim 1 , ethanol claim ...

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Номер записи: 98
19-09-2019 дата публикации

GERANYLGERANYL PYROPHOSPHATE SYNTHASES

Номер: US20190284539A1
Автор: BOER Viktor Marius, VAN LEEUWEN Johannes Gustaaf Ernst, ZWARTJENS Priscilla
Принадлежит: DSM IP ASSETS B.V.

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 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 positions92, 100 or 235said 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.2. A variant polypeptide according to claim 1 , wherein the modified property is modified geranylgeranyl pyrophosphate synthase activity.3. A variant polypeptide according to claim 1 , wherein the reference polypeptide comprises the geranylgeranyl pyrophosphate synthase of SEQ ID NO: 1 or SEQ ID NO: 17.4. A variant polypeptide according to claim 1 , wherein the variant polypeptide is a non-naturally occurring polypeptide.5. A variant polypeptide according to claim 1 , which comprises additional substitutions other than 92 claim 1 , 100 or 235.6. A variant polypeptide according to claim 1 , having at least 70% claim 1 , at least 75% claim 1 , at least ...

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Номер записи: 99
10-09-2020 дата публикации

TRANSFORMED PLANT HAVING BLUE FLOWER COLOR, AND METHOD FOR CREATING SAME

Номер: US20200283782A1
Автор: Nakamura Noriko, Okitsu Naoko, Tanaka Yoshikazu
Принадлежит: SUNTORY HOLDINGS LIMITED

Provided are a transformed plant having a blue-hued flower color or its inbred or outbred progenies, or their propagules, partial plant bodies, tissues or cells. 1. A method for creating a transformed plant characterized in that delphinidin-type anthocyanin and flavone C-glycoside are made to coexist in a cell of a plant.2. The method according to claim 1 , wherein the flavone C-glycoside is selected from the group consisting of flavone 6-C-glucoside claim 1 , flavone 8-C-glucoside claim 1 , and a combination thereof.3. The method according to claim 2 , wherein the flavone C-glycoside is apigenin 6-C-glucoside and/or luteolin 6-C-glucoside.4. The method according to claim 1 , wherein the delphinidin-type anthocyanin is selected from the group consisting of malvidin 3 claim 1 ,5-diglucoside claim 1 , delphinidin 3 claim 1 ,5-diglucoside claim 1 , petunidin 3 claim 1 ,5-diglucoside claim 1 , acylated anthocyanins claim 1 , and combinations thereof.5. The method according to comprising transforming a host plant with a vector comprising a flavone synthase (FNS) gene or a homologue thereof and a flavone C-glucosyltransferase (CGT) gene or a homologue thereof.6. The method according to claim 5 , wherein the vector further comprises a flavonoid 3′ claim 5 ,5′-hydroxylase (F3′5′H) gene or a homologue thereof and a methyltransferase (MT) gene or a homologue thereof.7. The method according to claim 6 , wherein:the FNS gene or homologue thereof is selected from the group consisting of(1-a) a polynucleotide consisting of the base sequence of SEQ ID NO: 19,(1-b) a polynucleotide hybridizing to a polynucleotide consisting of a base sequence complimentary to the base sequence of SEQ ID NO:19 under stringent conditions and encoding a protein having the same activity as a protein encoded by the polynucleotide set forth in (1-a),(1-c) a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO: 20,(1-d) a polynucleotide encoding a protein consisting of an ...

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Номер записи: 100