CN102260693A - 使用腈水解酶突变体生产3-羟基羧酸 - Google Patents
使用腈水解酶突变体生产3-羟基羧酸 Download PDFInfo
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- CN102260693A CN102260693A CN2011101955321A CN201110195532A CN102260693A CN 102260693 A CN102260693 A CN 102260693A CN 2011101955321 A CN2011101955321 A CN 2011101955321A CN 201110195532 A CN201110195532 A CN 201110195532A CN 102260693 A CN102260693 A CN 102260693A
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- 150000007517 lewis acids Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
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- 229910052749 magnesium Inorganic materials 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical compound O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- PSGAAPLEWMOORI-PEINSRQWSA-N medroxyprogesterone acetate Chemical compound C([C@@]12C)CC(=O)C=C1[C@@H](C)C[C@@H]1[C@@H]2CC[C@]2(C)[C@@](OC(C)=O)(C(C)=O)CC[C@H]21 PSGAAPLEWMOORI-PEINSRQWSA-N 0.000 description 1
- BRMYZIKAHFEUFJ-UHFFFAOYSA-L mercury diacetate Chemical compound CC(=O)O[Hg]OC(C)=O BRMYZIKAHFEUFJ-UHFFFAOYSA-L 0.000 description 1
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- 235000013379 molasses Nutrition 0.000 description 1
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- 239000002052 molecular layer Substances 0.000 description 1
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- 239000012074 organic phase Substances 0.000 description 1
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- 230000008520 organization Effects 0.000 description 1
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- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
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- 102200008576 rs121917899 Human genes 0.000 description 1
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- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
- 229910052718 tin Inorganic materials 0.000 description 1
- 239000011135 tin Substances 0.000 description 1
- XOLBLPGZBRYERU-UHFFFAOYSA-N tin dioxide Chemical compound O=[Sn]=O XOLBLPGZBRYERU-UHFFFAOYSA-N 0.000 description 1
- 229910001887 tin oxide Inorganic materials 0.000 description 1
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- 239000010936 titanium Substances 0.000 description 1
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- VOITXYVAKOUIBA-UHFFFAOYSA-N triethylaluminium Chemical compound CC[Al](CC)CC VOITXYVAKOUIBA-UHFFFAOYSA-N 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
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Classifications
-
- 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
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/42—Hydroxy-carboxylic acids
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
OD(600nm) | OD(600nm) | 3-HVN U/g | |
烧瓶 | 0h | 8h | 细胞干重 |
top10/pNM18-1 | 0.080 | 2.25 | 2.00 |
top10/pNM18-2 | 0.081 | 2.19 | 2.20 |
top10/pNM18-3 | 0.083 | 2.28 | 2.07 |
top10/pNM18-4 | 0.082 | 2.29 | 1.96 |
top10/pNM18-5 | 0.085 | 2.23 | 2.13 |
top10/pNM18-6 | 0.081 | 2.19 | 2.11 |
top10/pNM18(平均) | 2.08 |
OD(600nm) | OD(600nm) | 3-HVN U/g | |
烧瓶 | 0h | 8h | 细胞干重 |
top10/pNM18/B2-1 | 0.086 | 1.82 | 13.62 |
top10/pNM18/B2-2 | 0.073 | 1.89 | 11.47 |
top 10/pNM18/B2-3 | 0.088 | 2.13 | 12.83 |
top10/pNM18/B2-4 | 0.078 | 2.13 | 10.51 |
top10/pNM18/B2-5 | 0.085 | 2.13 | 11.67 |
top10/pNM18/B2-6 | 0.083 | 1.78 | 11.82 |
top10/pNM18/B2(平均) | 12.0 |
OD(600nm) | OD(600nm) | 3-HVN U/g | |
烧瓶 | 0h | 8h | 细胞干重 |
top10/pNM18/H9-1 | 0.095 | 2.89 | 16.32 |
top10/pNM18/H9-2 | 0.074 | 2.56 | 15.62 |
top10/pNM18/H9-3 | 0.090 | 2.51 | 15.84 |
top10/pNM18/H9-4 | 0.090 | 2.44 | 15.41 |
top10/pNM18/H9-5 | 0.100 | 2.46 | 14.41 |
top10/pNM18/H9-6 | 0.085 | 4.63 | 14.87 |
top10/pNM18/H9(平均) | 15.4 |
3-HVN U/g | 腈水解酶活性,相对于 | |
菌株 | 细胞干重 | top10/pNM18 |
top10/pNM18(对照) | 2.08 | 1.0 |
top10/pNM18/B4 | 3.67 | 1.8 |
top10/pNM18/B2 | 12.0 | 5.8 |
top10/pNM18/H9 | 15.4 | 7.4 |
菌株 | 3-HVN U/g dcw | 相对的腈水解酶活性 |
top10/pNM18(对照) | 1.77 | 1.0 |
top10/pNM18/210Cys | 11.0 | 6.2 |
化学物质 | 浓度 |
柠檬酸 | 10g/L |
CaCl2*2H2O | 1.5g/L |
FeSO4*7H2O | 5g/L |
ZnSO4*7H2O | 0.39g/L |
CuSO4*5H2O | 0.38g/L |
CoCl2*6H2O | 0.2g/L |
MnCl2*4H2O | 0.3g/L |
初始设定值 | 最小 | 最大 | |
搅拌器(rpm) | 400 | 400 | 850 |
气流(slpm) | 2 | 2 | 16 |
pH | 6.8 | 6.8 | 6.8 |
压力(kPa) | 3.45 | 3.45 | 3.45 |
DO | 25% | 25% | 25% |
温度℃ | 36 | 36 | 36 |
时间(h) | 速率(g/min) |
0-4 | 0.27 |
4-16 | 0.55 |
16-End | 0.42 |
底物 | U/g | |
菌株 | (0.5M) | 细胞干重 |
大肠杆菌top10/pNM18(对照) | 3-HVN | 1.94 |
大肠杆菌top10/pNM18(对照) | 3-HBN | 2.69 |
大肠杆菌top10/pNM18/H9 | 3-HVN | 14.0 |
大肠杆菌top10/pNM18/H9 | 3-HBN | 5.2 |
Claims (10)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/919,182 US7148051B2 (en) | 2004-08-16 | 2004-08-16 | Production of 3-hydroxycarboxylic acid using nitrilase |
US10/919182 | 2004-08-16 |
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CN2005800353017A Division CN101072871B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
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CN102260693A true CN102260693A (zh) | 2011-11-30 |
CN102260693B CN102260693B (zh) | 2013-06-19 |
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CN2011101955891A Active CN102260696B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
CN2011101955321A Active CN102260693B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
CN201110195511.XA Active CN102277363B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
CN2011101955482A Active CN102260695B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
CN201110195560.3A Active CN102286500B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
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CN2011101955891A Active CN102260696B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
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CN2011101955482A Active CN102260695B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
CN201110195560.3A Active CN102286500B (zh) | 2004-08-16 | 2005-08-16 | 使用腈水解酶突变体生产3-羟基羧酸 |
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US (3) | US7148051B2 (zh) |
EP (1) | EP1778840B1 (zh) |
JP (1) | JP4934037B2 (zh) |
CN (6) | CN101072871B (zh) |
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KR100250830B1 (ko) | 1997-12-09 | 2000-04-01 | 성재갑 | 자가분해에 의해 폴리하이드록시알킨산으로부터 광학활성을 가진 단량체 하이드록시카르복실산의 제조방법 |
EP1142997B1 (en) * | 1999-10-26 | 2009-12-02 | Showa Denko K.K. | Novel rhodococcus, rhodococcus-origin nitrilase gene, nitrilehydratase gene and amidase gene and process for producing carboxylic acids by using the same |
DE10010149A1 (de) * | 2000-03-03 | 2001-09-06 | Basf Ag | Nitrilase aus Rhodococcus rhodochrous NCIMB 11216 |
EP1280892B1 (en) | 2000-03-31 | 2011-01-12 | E.I. Du Pont De Nemours And Company | ISOLATION AND EXPRESSION OF A GENE FOR A NITRILASE FROM i ACIDOVORAX FACILIS 72W |
US6562603B2 (en) * | 2000-08-04 | 2003-05-13 | E. I. Du Pont De Nemours And Company | 3-hydroxycarboxylic acid production and use in branched polymers |
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US6416980B1 (en) * | 2001-02-23 | 2002-07-09 | E. I. Du Pont De Nemours & Company | Method for producing glycolic acid from glycolonitrile using nitrilase |
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2004
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2005
- 2005-08-16 DE DE602005007499T patent/DE602005007499D1/de active Active
- 2005-08-16 CN CN2005800353017A patent/CN101072871B/zh active Active
- 2005-08-16 CN CN2011101955891A patent/CN102260696B/zh active Active
- 2005-08-16 JP JP2007527960A patent/JP4934037B2/ja active Active
- 2005-08-16 CN CN2011101955321A patent/CN102260693B/zh active Active
- 2005-08-16 EP EP05787796A patent/EP1778840B1/en active Active
- 2005-08-16 WO PCT/US2005/029188 patent/WO2006023520A2/en active Application Filing
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Also Published As
Publication number | Publication date |
---|---|
CN102260693B (zh) | 2013-06-19 |
CN102260695A (zh) | 2011-11-30 |
EP1778840B1 (en) | 2008-06-11 |
DE602005007499D1 (de) | 2008-07-24 |
CN102286500B (zh) | 2014-02-19 |
US20100143989A1 (en) | 2010-06-10 |
CN101072871A (zh) | 2007-11-14 |
JP2008509708A (ja) | 2008-04-03 |
JP4934037B2 (ja) | 2012-05-16 |
CN101072871B (zh) | 2011-08-31 |
CN102277363A (zh) | 2011-12-14 |
CN102260696B (zh) | 2013-03-27 |
CN102260696A (zh) | 2011-11-30 |
US20060035352A1 (en) | 2006-02-16 |
US7148051B2 (en) | 2006-12-12 |
CN102286500A (zh) | 2011-12-21 |
WO2006023520A3 (en) | 2006-06-15 |
US7358071B2 (en) | 2008-04-15 |
WO2006023520A2 (en) | 2006-03-02 |
CN102260695B (zh) | 2012-09-05 |
CN102277363B (zh) | 2015-09-02 |
US20070117192A1 (en) | 2007-05-24 |
EP1778840A2 (en) | 2007-05-02 |
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