KR100359171B1 - 폴리하이드록시알칸산 생합성 효소와 세포내폴리하이드록시알칸산 분해효소를 발현시키는 재조합미생물 및 그를 이용한 (r)-3-하이드록시카르복실산의제조방법 - Google Patents
폴리하이드록시알칸산 생합성 효소와 세포내폴리하이드록시알칸산 분해효소를 발현시키는 재조합미생물 및 그를 이용한 (r)-3-하이드록시카르복실산의제조방법 Download PDFInfo
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- KR100359171B1 KR100359171B1 KR1020000026158A KR20000026158A KR100359171B1 KR 100359171 B1 KR100359171 B1 KR 100359171B1 KR 1020000026158 A KR1020000026158 A KR 1020000026158A KR 20000026158 A KR20000026158 A KR 20000026158A KR 100359171 B1 KR100359171 B1 KR 100359171B1
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Abstract
Description
플라스미드/배지 | 배양온도/배양시간 | 건조균체의농도(g/L) | PHB의농도(g/L) | PHB의함유량(%) | 단량체의농도(g/L) | 이량체의농도(g/L) | 최종수율(%) |
pJC4Red/R+포도당+티아민 | 37℃/51시간 | 1.45 | 0.10 | 7 | 0.7 | 6.7 | 43 |
pSYL105Red/R+포도당+티아민 | 37℃/51시간 | 1.50 | 0.47 | 31 | 1.6 | 6.1 | 44 |
pSYL107Red/R+포도당+티아민 | 30℃/51시간 | 3.04 | 1.97 | 65 | 1.7 | 2.7 | 25 |
pJC4Red/LB+포도당 | 37℃/51시간 | 2.72 | 1.20 | 44 | 0.2 | 2.6 | 16 |
pSYL105Red/LB+포도당 | 37℃/51시간 | 2.75 | 1.32 | 48 | 0.2 | 4.0 | 25 |
pSYL107Red/LB+포도당 | 30℃/51시간 | 3.84 | 2.81 | 73 | 0.9 | 0.3 | 6 |
플라스미드/배양온도 | 유도발현 후배양시간 | 건조균체의농도(g/L) | PHB의농도(g/L) | PHB의함유량(%) | 단량체의농도(g/L) | 이량체의농도(g/L) | 최종수율(%) |
pJC4Red-trc/37℃ | 2 시간 | 1.4 | 0.16 | 11 | 0.1 | 3.6 | 22 |
4 시간 | 1.2 | 0.09 | 8 | 0.5 | 4.3 | 28 | |
6 시간 | 1.1 | 0.09 | 8 | 0.5 | 5.0 | 32 | |
pSYL105Red-trc/37℃ | 2 시간 | 1.2 | 0.04 | 3 | 0.7 | 6.0 | 39 |
4 시간 | 1.3 | 0.00 | 0 | 0.7 | 6.1 | 40 | |
6 시간 | 1.3 | 0.02 | 2 | 0.7 | 7.1 | 45 | |
pSYL107Red-trc/30℃ | 2 시간 | 0.7 | 0.12 | 17 | 0.0 | 0.6 | 4 |
4 시간 | 0.8 | 0.15 | 19 | 0.1 | 1.2 | 8 | |
6 시간 | 0.8 | 0.15 | 19 | 0.3 | 1.5 | 10 |
재조합 대장균 | 배양온도/유도발현 후배양시간 | 건조균체의농도(g/L) | (R)-3-하이드록시부탄산의 농도(g/L) | (R)-3-하이드록시발레르산의농도(g/L) |
XL1-Blue/pJC4Red | 37℃/0시간 | 2.15 | 0.12 | 0.01 |
XL1-Blue/pSYL107Red | 30℃/0시간 | 1.10 | 0.10 | 0.00 |
XL1-Blue/pJC4Red-trc | 37℃/4시간 | 0.80 | 1.51 | 0.28 |
XL1-Blue/pSYL107Red-trc | 37℃/4시간 | 0.95 | 0.17 | 0.03 |
Claims (12)
- 폴리하이드록시알칸산(polyhydroxyalkanoate, PHA) 생합성효소 유전자와 PHA 분해효소 유전자를 포함하는 재조합 플라스미드.
- 제 1항에 있어서,폴리하이드록시알칸산 생합성효소 유전자는 알칼리게네스레이터스(Alcaligenes latus) 또는 랄스토니아 유트로파(Ralstoniaeutropha)로부터 유래한 것을 특징으로 하는재조합 플라스미드.
- 제 1항에 있어서,폴리하이드록시알칸산 분해효소 유전자는 랄스토니아유트로파(Ralstonia eutropha)로부터 유래된 것을 특징으로 하는재조합 플라스미드.
- 제 1항에 있어서,폴리하이드록시알칸산 분해효소 유전자의 발현 프로모터는 랄스토니아유트로파(Ralstonia eutropha) 고유의 발현 프로모터, trc, T7, trp.tac 및 bad 유도발현 프로모터로 구성된 그룹으로부터 선택되는것을 특징으로 하는재조합 플라스미드.
- 제 1항에 있어서,pJC4Red, pSYL105Red, pSYL107Red, pJC4Red-trc, pSYL105Red-trc 및pSYL107Red-trc로 부터 선택되는 것을 특징으로 하는재조합 플라스미드.
- 재조합 플라스미드 pJC4Red로 형질전환된 대장균 XL1-Blue/pJC4Red(Escherichia coliXL1-Blue/pJC4Red)(KCTC 0677BP).
- 재조합 플라스미드 pSYL105Red로 형질전환된 대장균 XL1-Blue/pSYL105Red(Escherichia coliXL1-Blue/pSYL105Red)(KCTC 0676BP).
- 재조합 플라스미드 pSYL105Red-trc로 형질전환된 대장균 XL1-Blue/pSYL105Red-trc(Escherichia coliXL1-Blue/pSYL105Red-trc)(KCTC 0678BP).
- 제 1항의 재조합 플라스미드로 형질전환된 대장균을 배양하여 균체 내에서 폴리하이드록시알칸산의 합성과 분해가 동시에 이루어지게 하고, 이로부터 (R)-3-하이드록시카르복실산을 수득하는 단계를 포함하는, 광학적으로 순수한 (R)-3-하이드록시카르복실산의 제조방법.
- 제 9항에 있어서,재조합 플라스미드는 pJC4Red, pSYL105Red, pSYL107Red, pJC4Red-trc,pSYL105Red-trc 및 pSYL107Red-trc로 구성된 그룹으로부터 선택되는것을 특징으로 하는광학적으로 순수한 (R)-3-하이드록시카르복실산의 제조방법.
- 제 9항에 있어서,형질전환된 대장균은 XL1-Blue/pJC4Red(Escherichia coliXL1-Blue/pJC4Red, KCTC 0677BP), XL1-Blue/pSYL105Red(Escherichia coliXL1-Blue/pSYL105Red, KCTC 0676BP) 또는 XL1-Blue/pSYL105Red-trc(Escherichia coliXL1-Blue/pSYL105Red-trc, KCTC 0678BP)인것을 특징으로 하는광학적으로 순수한 (R)-3-하이드록시카르복실산의 제조방법.
- 제 9항에 있어서,(R)-3-하이드록시카르복실산은 (R)-3-하이드록시부탄산, (R)-3-하이드록시발레르산, (R)-3-하이드록시부탄산의 이량체, (R)-3-하이드록시발레르산의 이량체, (R)-3-하이드록시부탄산, (R)-3-하이드록시발레르산의 에스터 또는 이들의 혼합물인 것을 특징으로 하는광학적으로 순수한 (R)-3-하이드록시카르복실산의 제조방법.
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020000026158A KR100359171B1 (ko) | 2000-05-16 | 2000-05-16 | 폴리하이드록시알칸산 생합성 효소와 세포내폴리하이드록시알칸산 분해효소를 발현시키는 재조합미생물 및 그를 이용한 (r)-3-하이드록시카르복실산의제조방법 |
PCT/KR2000/000787 WO2001088145A1 (en) | 2000-05-16 | 2000-07-20 | Recombinant microorganism expressing polyhydroxyalkanoate biosynthesis enzyme and intracellular pha depolymerase |
JP2001584527A JP2004516004A (ja) | 2000-05-16 | 2000-07-20 | ポリヒドロキシアルカノエート生合成酵素及び細胞内phaデポリメラーゼを発現する組換え微生物 |
EP00946504A EP1285070A4 (en) | 2000-05-16 | 2000-07-20 | RECOMBINANT MICROORGANISMS THAT EXPRESS AN ENZYME OF POLYHYDROXYALKANOAT BIOSYNTHESIS AND INTRACELLULAR PHA-DEPOLYMERASE |
CNB008195390A CN1246460C (zh) | 2000-05-16 | 2000-07-20 | 表达聚羟基烷酸生物合成酶和胞内pha解聚酶的重组微生物 |
US10/299,025 US20030143703A1 (en) | 2000-05-16 | 2002-11-15 | Recombinant microorganism expressing polyhydroxyalkanoate biosynthesis enzyme and intracellular PHA depolymerase |
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KR1020000026158A KR100359171B1 (ko) | 2000-05-16 | 2000-05-16 | 폴리하이드록시알칸산 생합성 효소와 세포내폴리하이드록시알칸산 분해효소를 발현시키는 재조합미생물 및 그를 이용한 (r)-3-하이드록시카르복실산의제조방법 |
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KR20010104925A KR20010104925A (ko) | 2001-11-28 |
KR100359171B1 true KR100359171B1 (ko) | 2002-10-31 |
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KR1020000026158A KR100359171B1 (ko) | 2000-05-16 | 2000-05-16 | 폴리하이드록시알칸산 생합성 효소와 세포내폴리하이드록시알칸산 분해효소를 발현시키는 재조합미생물 및 그를 이용한 (r)-3-하이드록시카르복실산의제조방법 |
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US (1) | US20030143703A1 (ko) |
EP (1) | EP1285070A4 (ko) |
JP (1) | JP2004516004A (ko) |
KR (1) | KR100359171B1 (ko) |
CN (1) | CN1246460C (ko) |
WO (1) | WO2001088145A1 (ko) |
Families Citing this family (12)
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KR100447532B1 (ko) * | 2001-11-28 | 2004-09-08 | 한국과학기술원 | (알)-하이드록시카르복실산 생산 재조합 미생물 및 그를이용한 (알)-하이드록시카르복실산의 제조방법 |
WO2008113190A1 (en) * | 2007-03-22 | 2008-09-25 | Empa Eidgenössische Materialprüfungs- Und Forschungsanstalt | Method for the production of r-hydroxycarboxylic acids |
KR101114918B1 (ko) * | 2007-08-09 | 2012-02-15 | 주식회사 엘지화학 | 재조합 미생물을 이용한 광학활성(s)-3-하이드록시부탄산 및(s)-3-하이드록시부티레이트 에스테르의 제조방법 |
DE102008002715A1 (de) * | 2008-06-27 | 2009-12-31 | Evonik Röhm Gmbh | 2-Hydroxyisobuttersäure produzierende rekombinante Zelle |
SG181607A1 (en) * | 2009-12-10 | 2012-07-30 | Genomatica Inc | Methods and organisms for converting synthesis gas or other gaseous carbon sources and methanol to 1,3-butanediol |
CN103275915B (zh) * | 2013-06-05 | 2014-12-03 | 中国农业大学 | 一株产聚羟基烷酸重组突变菌株 |
WO2015097104A1 (en) * | 2013-12-23 | 2015-07-02 | Carbios | Method for recycling plastic products |
CN107299072B (zh) * | 2017-08-02 | 2020-11-06 | 江南大学 | 一种工程菌及其应用 |
CN109722444B (zh) * | 2019-01-02 | 2020-11-13 | 齐鲁工业大学 | 一种重组质粒pZQ12、合成奇偶数碳链单体共聚mcl-PHA的重组菌及其应用 |
CN110904161A (zh) * | 2019-12-27 | 2020-03-24 | 浙江英玛特生物科技有限公司 | 一种采用酶法生产高纯度(r)-(-)-3-羟基丁酸的方法 |
KR20230169409A (ko) * | 2021-05-28 | 2023-12-15 | 킴벌리-클라크 월드와이드, 인크. | 산업 응용을 위한 호염성 phb 탈중합효소의 최적화 |
AU2022280785A1 (en) * | 2021-05-28 | 2024-01-04 | Kimberly-Clark Worldwide, Inc. | Methods and systems for single-step decontamination and enzymatic degradation of bio-based polymers |
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US5480794A (en) * | 1987-06-29 | 1996-01-02 | Massachusetts Institute Of Technology And Metabolix, Inc. | Overproduction and purification of soluble PHA synthase |
FR2641532B1 (fr) * | 1989-01-06 | 1991-03-29 | Solvay | Procede pour la preparation d'esters de l'acide (beta)-hydroxybutyrique |
DE4209616A1 (de) * | 1992-03-25 | 1993-09-30 | Basf Ag | Verfahren zur Herstellung von µ-Hydroxycarbonsäureestern |
JPH09234091A (ja) * | 1995-12-28 | 1997-09-09 | Akira Shimizu | 発酵法によるR−β−ヒドロキシ酪酸の製造法 |
KR100250830B1 (ko) * | 1997-12-09 | 2000-04-01 | 성재갑 | 자가분해에 의해 폴리하이드록시알킨산으로부터 광학활성을 가진 단량체 하이드록시카르복실산의 제조방법 |
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2000
- 2000-05-16 KR KR1020000026158A patent/KR100359171B1/ko active IP Right Grant
- 2000-07-20 EP EP00946504A patent/EP1285070A4/en not_active Withdrawn
- 2000-07-20 JP JP2001584527A patent/JP2004516004A/ja active Pending
- 2000-07-20 WO PCT/KR2000/000787 patent/WO2001088145A1/en not_active Application Discontinuation
- 2000-07-20 CN CNB008195390A patent/CN1246460C/zh not_active Expired - Fee Related
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2002
- 2002-11-15 US US10/299,025 patent/US20030143703A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
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KR20010104925A (ko) | 2001-11-28 |
US20030143703A1 (en) | 2003-07-31 |
EP1285070A1 (en) | 2003-02-26 |
WO2001088145A1 (en) | 2001-11-22 |
CN1246460C (zh) | 2006-03-22 |
EP1285070A4 (en) | 2004-07-28 |
CN1457364A (zh) | 2003-11-19 |
JP2004516004A (ja) | 2004-06-03 |
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