RU2005134209A - Микроорганизм с инактивированными генами tdcbc/pcka и способ получения l-треонина с его использованием - Google Patents
Микроорганизм с инактивированными генами tdcbc/pcka и способ получения l-треонина с его использованием Download PDFInfo
- Publication number
- RU2005134209A RU2005134209A RU2005134209/13A RU2005134209A RU2005134209A RU 2005134209 A RU2005134209 A RU 2005134209A RU 2005134209/13 A RU2005134209/13 A RU 2005134209/13A RU 2005134209 A RU2005134209 A RU 2005134209A RU 2005134209 A RU2005134209 A RU 2005134209A
- Authority
- RU
- Russia
- Prior art keywords
- gene
- pcka
- escherichia coli
- inactivated
- threonine
- Prior art date
Links
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 title claims abstract 21
- 108090000623 proteins and genes Proteins 0.000 title claims abstract 11
- 244000005700 microbiome Species 0.000 title abstract 6
- 238000004519 manufacturing process Methods 0.000 title abstract 2
- 101150088738 pckA gene Proteins 0.000 claims abstract 16
- 239000004473 Threonine Substances 0.000 claims abstract 10
- 229960002898 threonine Drugs 0.000 claims abstract 10
- 230000003115 biocidal effect Effects 0.000 claims abstract 4
- 230000015556 catabolic process Effects 0.000 claims abstract 4
- 230000002759 chromosomal effect Effects 0.000 claims abstract 4
- 238000006731 degradation reaction Methods 0.000 claims abstract 4
- 210000000349 chromosome Anatomy 0.000 claims abstract 3
- 238000000034 method Methods 0.000 claims abstract 3
- 239000003242 anti bacterial agent Substances 0.000 claims abstract 2
- 230000000694 effects Effects 0.000 claims abstract 2
- 241000588724 Escherichia coli Species 0.000 claims 10
- 239000012634 fragment Substances 0.000 claims 6
- 239000013612 plasmid Substances 0.000 claims 5
- 108010052160 Site-specific recombinase Proteins 0.000 claims 4
- 101150067708 pckG gene Proteins 0.000 claims 3
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 claims 2
- 229960005091 chloramphenicol Drugs 0.000 claims 2
- 238000010367 cloning Methods 0.000 claims 1
- 230000006801 homologous recombination Effects 0.000 claims 1
- 238000002744 homologous recombination Methods 0.000 claims 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 abstract 1
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 239000008103 glucose Substances 0.000 abstract 1
- 230000002779 inactivation Effects 0.000 abstract 1
- 230000004941 influx Effects 0.000 abstract 1
- 230000003834 intracellular effect Effects 0.000 abstract 1
- 230000037361 pathway Effects 0.000 abstract 1
- 238000005215 recombination Methods 0.000 abstract 1
- 230000006798 recombination Effects 0.000 abstract 1
Classifications
-
- 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
-
- 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
- C12N1/20—Bacteria; 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Claims (7)
1. Штамм Escherichia coli, содержащий оба гена, tdcBC и pckA, которые инактивированы.
2. Штамм Escherichia coli по п.1, в котором pckA-ген инактивируют путем встраивания фрагмента чужеродного pckA-гена, содержащего устойчивый к антибиотику ген, имеющий сайт связывания сайт-специфической рекомбиназы на каждом его конце в родительском штамме Escherichia coli, содержащем ассоциированный с деградацией L-треонина оперон tdcBC, который инактивирован, а затем гомологической рекомбинации между фрагментом чужеродного pckA-гена и pckA-геном на хромосоме для инактивации хромосомного pckA-гена.
3. Штамм Escherichia coli по п.2, в котором pckA-ген инактивируют путем удаления встроенного в него устойчивого к антибиотику гена в результате активности сайт-специфической рекомбиназы, экспрессируемой в штамме Escherichia coli, и наличия одной копии связывающего сайта сайт-специфической рекомбиназы в хромосомном pckA-гене.
4. Штамм Escherichia coli по п.2, в котором сайт-специфическая рекомбиназа представляет собой FLP, Cre или XerC/D.
5. Штамм Escherichia coli по п.2, представляющий собой Escherichia coli FTR2717 (KCCM-10475), содержащий на хромосоме pckA-ген, инактивируемый путем введения фрагмента чужеродного pckA-гена, содержащего устойчивый к антибиотику ген, имеющий loxP-сайт на каждом его конце в родительском штамме Escherichia coli, содержащем ассоциированный с деградацией L-треонина tdcBC-оперон, который инактивирован.
6. Способ получения L-треонина с использованием штамма Escherichia coli по любому из пп.1-5.
7. Рекомбинантная плазмида pT7ΔpckA::loxpcat, содержащая фрагмент pckA-гена, содержащий устойчивый к хлорамфениколу ген и loxP-сайты, где указанную рекомбинантную плазмиду получают путем клонирования неполного pckA-гена в pT7Blue-вектор, с образованием pT7Blue/pckA-плазмиды, получая из ploxpcat2-плазмиды ДНК-фрагмент, содержащий устойчивый к хлорамфениколу ген и loxP-сайты, loxpcat2, и лигируя loxpcat-ДНК-фрагмент с NruI-обработанной pT7Blue/pckA-плазмидой.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2003-0021458 | 2003-04-04 | ||
| KR10-2003-0021458A KR100498971B1 (ko) | 2003-04-04 | 2003-04-04 | 염색체 내 ｔｄｃＢＣ 및 ｐｃｋＡ 유전자가 불활성화된 미생물 및 이를 이용하여 Ｌ-쓰레오닌을 제조하는 방법 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| RU2005134209A true RU2005134209A (ru) | 2006-03-20 |
| RU2339699C2 RU2339699C2 (ru) | 2008-11-27 |
Family
ID=36117119
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2005134209/13A RU2339699C2 (ru) | 2003-04-04 | 2004-04-02 | РЕКОМБИНАНТНАЯ ПЛАЗМИДА pT7ΔpckA::loxpcat ОБЕСПЕЧИВАЮЩАЯ СИНТЕЗ L-ТРЕОНИНА В КЛЕТКАХ Escherichia coli, И РЕКОМБИНАНТНЫЙ ШТАММ Escherichia coli FTR2717 (KCCM-10475) - ПРОДУЦЕНТ L-ТРЕОНИНА |
Country Status (17)
| Country | Link |
|---|---|
| US (1) | US7485450B2 (ru) |
| EP (1) | EP1611232B1 (ru) |
| JP (1) | JP4334565B2 (ru) |
| KR (1) | KR100498971B1 (ru) |
| CN (1) | CN100372927C (ru) |
| AT (1) | ATE455844T1 (ru) |
| AU (1) | AU2004225550B2 (ru) |
| BR (1) | BRPI0409192A (ru) |
| DE (1) | DE602004025218D1 (ru) |
| ES (1) | ES2338555T3 (ru) |
| HU (1) | HUP0501100A3 (ru) |
| MX (1) | MXPA05010725A (ru) |
| PL (1) | PL380072A1 (ru) |
| RU (1) | RU2339699C2 (ru) |
| SK (1) | SK1192005A3 (ru) |
| WO (1) | WO2004087895A1 (ru) |
| ZA (1) | ZA200507688B (ru) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1735339A2 (en) * | 2004-04-09 | 2006-12-27 | Genencor International, Inc. | Pcka modifications and enhanced protein expression in bacillus |
| KR100858913B1 (ko) * | 2007-03-09 | 2008-09-17 | 한국과학기술원 | 부위특이적 변이에 의해 제조된 고수율의 ｌ－쓰레오닌생산균주 및 이를 이용한 ｌ－쓰레오닌의 제조방법 |
| KR101608734B1 (ko) * | 2014-03-21 | 2016-04-04 | 씨제이제일제당 주식회사 | L-아미노산을 생산하는 미생물 및 이를 이용하여 l-아미노산을 생산하는 방법 |
| CN116606785A (zh) * | 2022-02-08 | 2023-08-18 | 廊坊梅花生物技术开发有限公司 | 一种修饰的棒状杆菌属微生物及其应用与构建方法 |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU875663A1 (ru) * | 1978-06-30 | 1982-09-15 | Всесоюзный научно-исследовательский институт генетики и селекции промышленных микроорганизмов | Штаммы е.coLI ВНИИГенетика VL 334 @ N6 и ВНИИГенетика VL 334 @ N7-продуценты L-треонина и способ их получени |
| WO1995006114A1 (fr) * | 1993-08-24 | 1995-03-02 | Ajinomoto Co., Inc. | Allele de phosphenolpyruvate carboxylase, gene de cet allele et procede de production de l'acide amine |
| DE4400926C1 (de) * | 1994-01-14 | 1995-06-01 | Forschungszentrum Juelich Gmbh | Herstellung von L-Isoleucin mittels rekombinanter Mikroorganismen mit deregulierter Threonindehydratase |
| KR20010006976A (ko) | 1999-04-09 | 2001-01-26 | 김대훈 | 홍채인식시스템 |
| DE19950409A1 (de) * | 1999-10-20 | 2001-04-26 | Degussa | Neue für das pck-Gen codierende Nukleotidsequenzen |
| DE10109690A1 (de) * | 2000-09-02 | 2002-03-14 | Degussa | Neue für das metY-Gen kodierende Nukleotidsequenzen |
| AU2001293795A1 (en) * | 2000-09-30 | 2002-04-15 | Degussa A.G. | Fermentation process for the preparation of l-amino acids using strains of the family enterobacteriaceae |
| KR100397423B1 (ko) * | 2001-02-13 | 2003-09-13 | 씨제이 주식회사 | Ｌ-쓰레오닌의 제조방법 |
| DE10144493A1 (de) * | 2001-09-11 | 2003-07-03 | Degussa | Verfahren zur fermentativen Herstellung von L-Aminosäuren unter Verwendung coyneformer Bakterien |
| KR100451299B1 (ko) * | 2002-03-21 | 2004-10-06 | 씨제이 주식회사 | Ｌ―쓰레오닌의 제조방법 |
| KR100505797B1 (ko) * | 2002-10-11 | 2005-08-04 | 씨제이 주식회사 | 염색체 상의 ｆａｄＲ 유전자가 불활성화된 Ｌ-쓰레오닌 생성 변이 미생물과 이를 이용한 Ｌ-쓰레오닌의 제조방법 |
| DE102004049692A1 (de) | 2004-10-12 | 2006-04-20 | Siemens Ag | Vorrichtung zum Umschalten eines Telekommunikations-Endgerätes, Verwendungen und Verfahren |
-
2003
- 2003-04-04 KR KR10-2003-0021458A patent/KR100498971B1/ko active IP Right Grant
-
2004
- 2004-04-02 AU AU2004225550A patent/AU2004225550B2/en not_active Expired
- 2004-04-02 AT AT04725555T patent/ATE455844T1/de not_active IP Right Cessation
- 2004-04-02 WO PCT/KR2004/000778 patent/WO2004087895A1/en active Application Filing
- 2004-04-02 RU RU2005134209/13A patent/RU2339699C2/ru active
- 2004-04-02 PL PL380072A patent/PL380072A1/pl not_active Application Discontinuation
- 2004-04-02 CN CNB2004800086929A patent/CN100372927C/zh not_active Expired - Lifetime
- 2004-04-02 HU HU0501100A patent/HUP0501100A3/hu unknown
- 2004-04-02 JP JP2006500668A patent/JP4334565B2/ja not_active Expired - Lifetime
- 2004-04-02 BR BRPI0409192-2A patent/BRPI0409192A/pt not_active IP Right Cessation
- 2004-04-02 SK SK119-2005A patent/SK1192005A3/sk not_active Application Discontinuation
- 2004-04-02 EP EP04725555A patent/EP1611232B1/en not_active Expired - Lifetime
- 2004-04-02 MX MXPA05010725A patent/MXPA05010725A/es active IP Right Grant
- 2004-04-02 ES ES04725555T patent/ES2338555T3/es not_active Expired - Lifetime
- 2004-04-02 US US10/817,044 patent/US7485450B2/en active Active
- 2004-04-02 DE DE602004025218T patent/DE602004025218D1/de not_active Expired - Fee Related
-
2005
- 2005-09-23 ZA ZA200507688A patent/ZA200507688B/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| MXPA05010725A (es) | 2005-12-15 |
| HUP0501100A2 (en) | 2007-06-28 |
| AU2004225550B2 (en) | 2006-09-28 |
| EP1611232B1 (en) | 2010-01-20 |
| JP4334565B2 (ja) | 2009-09-30 |
| US7485450B2 (en) | 2009-02-03 |
| DE602004025218D1 (de) | 2010-03-11 |
| SK1192005A3 (sk) | 2006-03-02 |
| HUP0501100A3 (en) | 2010-01-28 |
| BRPI0409192A (pt) | 2006-04-11 |
| PL380072A1 (pl) | 2006-12-27 |
| WO2004087895A1 (en) | 2004-10-14 |
| CN1768132A (zh) | 2006-05-03 |
| ES2338555T3 (es) | 2010-05-10 |
| EP1611232A1 (en) | 2006-01-04 |
| AU2004225550A1 (en) | 2004-10-14 |
| CN100372927C (zh) | 2008-03-05 |
| ATE455844T1 (de) | 2010-02-15 |
| RU2339699C2 (ru) | 2008-11-27 |
| JP2006521789A (ja) | 2006-09-28 |
| KR100498971B1 (ko) | 2005-07-04 |
| ZA200507688B (en) | 2006-09-27 |
| KR20040087188A (ko) | 2004-10-13 |
| EP1611232A4 (en) | 2006-08-23 |
| US20040241831A1 (en) | 2004-12-02 |
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