AR086978A1 - Procedimiento de preparacion de levaduras geneticamente transformadas capaces de producir una molecula de interes con alta titulacion - Google Patents
Procedimiento de preparacion de levaduras geneticamente transformadas capaces de producir una molecula de interes con alta titulacionInfo
- Publication number
- AR086978A1 AR086978A1 ARP120102168A ARP120102168A AR086978A1 AR 086978 A1 AR086978 A1 AR 086978A1 AR P120102168 A ARP120102168 A AR P120102168A AR P120102168 A ARP120102168 A AR P120102168A AR 086978 A1 AR086978 A1 AR 086978A1
- Authority
- AR
- Argentina
- Prior art keywords
- yeast
- culture
- interest
- isolated
- expression cassettes
- Prior art date
Links
- 238000000034 method Methods 0.000 title abstract 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract 9
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 abstract 6
- 239000013612 plasmid Substances 0.000 abstract 4
- 229960000890 hydrocortisone Drugs 0.000 abstract 3
- 230000010354 integration Effects 0.000 abstract 3
- 239000003550 marker Substances 0.000 abstract 3
- 238000012216 screening Methods 0.000 abstract 3
- 102000003804 Adrenodoxin Human genes 0.000 abstract 2
- 108090000187 Adrenodoxin Proteins 0.000 abstract 2
- 101710088194 Dehydrogenase Proteins 0.000 abstract 1
- 101100054106 Sus scrofa HSD3B gene Proteins 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/79—Transferrins, e.g. lactoferrins, ovotransferrins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
- C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
- C12N9/0079—Steroid 11 beta monooxygenase (P-450 protein)(1.14.15.4)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
- C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
- C12N9/0081—Cholesterol monooxygenase (cytochrome P 450scc)(1.14.15.6)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
- C12Y101/01051—3 (or 17)-Beta-hydroxysteroid dehydrogenase (1.1.1.51)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Biophysics (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Virology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Reivindicación 1: Un procedimiento para preparar un cultivo aislado de levadura que produce hidrocortisona, que comprende: (a) proporcionar dos plásmidos de integración con cuatro casetes de expresión, comprendiendo cada plásmido de integración al menos dos casetes de expresión y opcionalmente un marcador seleccionable, en el que los cuatro casetes de expresión son P450scc, adrenodoxina (ADX), P450c11, y 3b-hidroxiesteroide deshidrogenasa (3b-HSD), (b) integrar de manera estable múltiples copias de los dos plásmidos de integración en una población de células de levadura, por co-transformación de los plásmidos en la levadura, (c) realizar un cribado primario para seleccionar al menos 30 clones de levadura, en el que la selección de los clones se basa en la presencia de los casetes de expresión, o la selección para la expresión del marcador seleccionable cuando dicho marcador está presente, y (d) realizar un cribado secundario funcional en los al menos 30 clones de levadura seleccionados en el cribado primario, para identificar un cultivo aislado de levadura que produce hidrocortisona.Reivindicación 15: Uso de un cultivo aislado de levadura preparado según el procedimiento de la reivindicación 1 para producir hidrocortisona. Reivindicación 34: Un procedimiento para producir una molécula de interés, que comprende: (a) preparar un cultivo aislado de levadura que produce una molécula de interés usando el procedimiento de la reivindicación 1; (b) cultivar el cultivo aislado de levadura en un medio de cultivo; y (c) recuperar y purificar dicha molécula de interés del cultivo.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1155462A FR2976949B1 (fr) | 2011-06-21 | 2011-06-21 | Procede de preparation de levures genetiquement transformees capables de produire une molecule d'interet a haut titre |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AR086978A1 true AR086978A1 (es) | 2014-02-05 |
Family
ID=46320970
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ARP120102168A AR086978A1 (es) | 2011-06-21 | 2012-06-19 | Procedimiento de preparacion de levaduras geneticamente transformadas capaces de producir una molecula de interes con alta titulacion |
Country Status (14)
| Country | Link |
|---|---|
| US (3) | US9994888B2 (es) |
| EP (1) | EP2723868B1 (es) |
| CN (1) | CN103797121B (es) |
| AR (1) | AR086978A1 (es) |
| AU (1) | AU2012271942B2 (es) |
| DK (1) | DK2723868T3 (es) |
| ES (1) | ES2639916T3 (es) |
| FR (1) | FR2976949B1 (es) |
| HU (1) | HUE034873T2 (es) |
| MY (1) | MY164066A (es) |
| NO (1) | NO2723868T3 (es) |
| TW (1) | TWI555843B (es) |
| UY (1) | UY34148A (es) |
| WO (1) | WO2012175453A1 (es) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2976949B1 (fr) * | 2011-06-21 | 2015-08-28 | Sanofi Sa | Procede de preparation de levures genetiquement transformees capables de produire une molecule d'interet a haut titre |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1042567A (zh) * | 1988-09-23 | 1990-05-30 | 吉斯特-布罗卡迪斯公司 | 类固醇多步氧化方法和所用的遗传工程细胞 |
| US5547868A (en) * | 1993-06-09 | 1996-08-20 | Regents Of The University Of California | Cholesterol disposal fusion enzymes |
| IL129722A0 (en) * | 1996-11-13 | 2000-02-29 | Du Pont | Method for the production of 1,3-propanediol by recombinant organisms |
| DE19919124A1 (de) * | 1999-04-27 | 2000-11-23 | Rhein Biotech Proz & Prod Gmbh | Nukleinsäuremolekül, umfassend eine für ein Polypeptid mit Chorismatmutase-Aktivität kodierende Nukleinsäure |
| FR2812884B1 (fr) * | 2000-08-08 | 2002-11-08 | Aventis Pharma Sa | Levures modifiees et utilisations, notamment pour la production de derives steroidiens |
| FR2820145B1 (fr) * | 2001-01-31 | 2004-01-23 | Aventis Pharma Sa | Souche de levure produisant des steroides de facon autonome |
| DE10312314A1 (de) * | 2003-03-19 | 2004-09-30 | Basf Ag | Verfahren zur Herstellung von Ergosta-5,7-dienol und/oder dessen biosynthetischen Zwischen- und/oder Folgeprodukten in transgenen Organismen |
| FR2869914B1 (fr) * | 2004-05-06 | 2012-11-09 | Aventis Pharma Sa | Souches de levure produisant du cholesterol et leurs applications |
| FR2976949B1 (fr) * | 2011-06-21 | 2015-08-28 | Sanofi Sa | Procede de preparation de levures genetiquement transformees capables de produire une molecule d'interet a haut titre |
-
2011
- 2011-06-21 FR FR1155462A patent/FR2976949B1/fr not_active Expired - Fee Related
-
2012
- 2012-06-18 DK DK12728522.9T patent/DK2723868T3/en active
- 2012-06-18 HU HUE12728522A patent/HUE034873T2/hu unknown
- 2012-06-18 CN CN201280041052.2A patent/CN103797121B/zh active Active
- 2012-06-18 EP EP12728522.9A patent/EP2723868B1/en active Active
- 2012-06-18 MY MYPI2013004523A patent/MY164066A/en unknown
- 2012-06-18 US US14/127,396 patent/US9994888B2/en active Active
- 2012-06-18 NO NO12728522A patent/NO2723868T3/no unknown
- 2012-06-18 AU AU2012271942A patent/AU2012271942B2/en not_active Ceased
- 2012-06-18 WO PCT/EP2012/061601 patent/WO2012175453A1/en not_active Ceased
- 2012-06-18 ES ES12728522.9T patent/ES2639916T3/es active Active
- 2012-06-19 AR ARP120102168A patent/AR086978A1/es not_active Application Discontinuation
- 2012-06-21 UY UY0001034148A patent/UY34148A/es not_active Application Discontinuation
- 2012-06-21 TW TW101122293A patent/TWI555843B/zh not_active IP Right Cessation
-
2018
- 2018-04-25 US US15/962,782 patent/US10400261B2/en active Active
-
2019
- 2019-07-22 US US16/518,607 patent/US20200048676A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| EP2723868A1 (en) | 2014-04-30 |
| DK2723868T3 (en) | 2017-10-30 |
| AU2012271942A1 (en) | 2013-05-02 |
| CN103797121B (zh) | 2018-04-10 |
| US10400261B2 (en) | 2019-09-03 |
| US9994888B2 (en) | 2018-06-12 |
| ES2639916T3 (es) | 2017-10-30 |
| CN103797121A (zh) | 2014-05-14 |
| MY164066A (en) | 2017-11-15 |
| US20140186885A1 (en) | 2014-07-03 |
| US20180245120A1 (en) | 2018-08-30 |
| AU2012271942B2 (en) | 2016-06-16 |
| HUE034873T2 (hu) | 2018-03-28 |
| EP2723868B1 (en) | 2017-07-26 |
| TWI555843B (zh) | 2016-11-01 |
| WO2012175453A1 (en) | 2012-12-27 |
| FR2976949B1 (fr) | 2015-08-28 |
| US20200048676A1 (en) | 2020-02-13 |
| FR2976949A1 (fr) | 2012-12-28 |
| NO2723868T3 (es) | 2017-12-23 |
| UY34148A (es) | 2013-01-31 |
| TW201305330A (zh) | 2013-02-01 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FA | Abandonment or withdrawal |