CN108271374B - 具有改进的活性的新颖p450-bm3变体 - Google Patents

具有改进的活性的新颖p450-bm3变体 Download PDF

Info

Publication number
CN108271374B
CN108271374B CN201680040014.3A CN201680040014A CN108271374B CN 108271374 B CN108271374 B CN 108271374B CN 201680040014 A CN201680040014 A CN 201680040014A CN 108271374 B CN108271374 B CN 108271374B
Authority
CN
China
Prior art keywords
leu
ala
glu
lys
asp
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201680040014.3A
Other languages
English (en)
Chinese (zh)
Other versions
CN108271374A (zh
Inventor
罗伯特·奥斯本
韦丝娜·米切尔
崔钦育奈
张希云
埃里卡·M·米尔泽克
杰弗里·C·穆尔
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Codexis Inc
Original Assignee
Codexis Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Codexis Inc filed Critical Codexis Inc
Publication of CN108271374A publication Critical patent/CN108271374A/zh
Application granted granted Critical
Publication of CN108271374B publication Critical patent/CN108271374B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
    • C12N9/0036—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on NADH or NADPH (1.6)
    • C12N9/0038—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on NADH or NADPH (1.6) with a heme protein as acceptor (1.6.2)
    • C12N9/0042—NADPH-cytochrome P450 reductase (1.6.2.4)
    • 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)
    • 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; 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
    • C12N1/00—Microorganisms; 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
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y106/00—Oxidoreductases acting on NADH or NADPH (1.6)
    • C12Y106/02—Oxidoreductases acting on NADH or NADPH (1.6) with a heme protein as acceptor (1.6.2)
    • C12Y106/02004—NADPH-hemoprotein reductase (1.6.2.4), i.e. NADP-cytochrome P450-reductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/14—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen (1.14.14)
    • C12Y114/14001—Unspecific monooxygenase (1.14.14.1)

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Botany (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
CN201680040014.3A 2015-07-07 2016-06-02 具有改进的活性的新颖p450-bm3变体 Active CN108271374B (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201562189281P 2015-07-07 2015-07-07
US62/189,281 2015-07-07
PCT/US2016/035399 WO2017007547A1 (en) 2015-07-07 2016-06-02 Novel p450-bm3 variants with improved activity

Publications (2)

Publication Number Publication Date
CN108271374A CN108271374A (zh) 2018-07-10
CN108271374B true CN108271374B (zh) 2021-09-21

Family

ID=57686072

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201680040014.3A Active CN108271374B (zh) 2015-07-07 2016-06-02 具有改进的活性的新颖p450-bm3变体

Country Status (8)

Country Link
US (7) US9683220B2 (https=)
EP (1) EP3319987B1 (https=)
JP (1) JP6691558B2 (https=)
CN (1) CN108271374B (https=)
CA (1) CA2991195C (https=)
DK (1) DK3319987T3 (https=)
IL (1) IL256441B (https=)
WO (1) WO2017007547A1 (https=)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9708587B2 (en) * 2014-07-09 2017-07-18 Codexis, Inc. P450-BM3 variants with improved activity
US9683220B2 (en) * 2015-07-07 2017-06-20 Codexis, Inc. P450-BM3 variants with improved activity
ES2892972T3 (es) * 2015-11-02 2022-02-07 Univ Texas Métodos de activación de CD40 y bloqueo de punto de control inmunitario
US11591627B2 (en) 2018-03-21 2023-02-28 University Of Florida Research Foundation, Incorporated Modified bacteria for production of nitroaromatics
EP4622999A1 (en) * 2022-11-22 2025-10-01 Codexis, Inc. P450-bm3 variants with improved activity
CN120813367A (zh) * 2023-03-20 2025-10-17 科德克希思公司 具有改进的活性的p450-bm3变体

Family Cites Families (61)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5834252A (en) 1995-04-18 1998-11-10 Glaxo Group Limited End-complementary polymerase reaction
US6117679A (en) 1994-02-17 2000-09-12 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6395547B1 (en) 1994-02-17 2002-05-28 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6995017B1 (en) 1994-02-17 2006-02-07 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US20060257890A1 (en) 1996-05-20 2006-11-16 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US5605793A (en) 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination
US5837458A (en) 1994-02-17 1998-11-17 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US6165793A (en) 1996-03-25 2000-12-26 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US5928905A (en) 1995-04-18 1999-07-27 Glaxo Group Limited End-complementary polymerase reaction
US6309883B1 (en) 1994-02-17 2001-10-30 Maxygen, Inc. Methods and compositions for cellular and metabolic engineering
US6335160B1 (en) 1995-02-17 2002-01-01 Maxygen, Inc. Methods and compositions for polypeptide engineering
US6406855B1 (en) 1994-02-17 2002-06-18 Maxygen, Inc. Methods and compositions for polypeptide engineering
FI964808A0 (fi) 1994-06-03 1996-12-02 Novo Nordisk Biotech Inc Puhdistetut Myceliophthora lakkaasit ja niitä koodaavat nukleiinihapot
CN101659926A (zh) 1994-06-30 2010-03-03 诺沃奇梅兹有限公司 非毒性、非产毒性、非致病性镰孢属表达系统及所用启动子和终止子
US5483345A (en) 1994-09-06 1996-01-09 Mrs Technology, Inc. Alignment system for use in lithography utilizing a spherical reflector having a centered etched-on projection object
FI104465B (fi) 1995-06-14 2000-02-15 Valio Oy Proteiinihydrolysaatteja allergioiden hoitamiseksi tai estämiseksi, niiden valmistus ja käyttö
GB2306485B (en) * 1995-11-01 1998-12-09 British Gas Plc Enzyme mutant
US6506602B1 (en) 1996-03-25 2003-01-14 Maxygen, Inc. Methods for generating polynucleotides having desired characteristics by iterative selection and recombination
US6096548A (en) 1996-03-25 2000-08-01 Maxygen, Inc. Method for directing evolution of a virus
US7148054B2 (en) 1997-01-17 2006-12-12 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
US6326204B1 (en) 1997-01-17 2001-12-04 Maxygen, Inc. Evolution of whole cells and organisms by recursive sequence recombination
KR100570935B1 (ko) 1997-01-17 2006-04-13 맥시겐, 인크. 반복적 서열 재조합에 의한 전세포 및 유기체의 개량 방법
US6537746B2 (en) 1997-12-08 2003-03-25 Maxygen, Inc. Method for creating polynucleotide and polypeptide sequences
US6365408B1 (en) 1998-06-19 2002-04-02 Maxygen, Inc. Methods of evolving a polynucleotides by mutagenesis and recombination
JP4221100B2 (ja) 1999-01-13 2009-02-12 エルピーダメモリ株式会社 半導体装置
US6376246B1 (en) 1999-02-05 2002-04-23 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US6368861B1 (en) 1999-01-19 2002-04-09 Maxygen, Inc. Oligonucleotide mediated nucleic acid recombination
US6917882B2 (en) 1999-01-19 2005-07-12 Maxygen, Inc. Methods for making character strings, polynucleotides and polypeptides having desired characteristics
US6436675B1 (en) 1999-09-28 2002-08-20 Maxygen, Inc. Use of codon-varied oligonucleotide synthesis for synthetic shuffling
US7024312B1 (en) 1999-01-19 2006-04-04 Maxygen, Inc. Methods for making character strings, polynucleotides and polypeptides having desired characteristics
US7702464B1 (en) 2001-08-21 2010-04-20 Maxygen, Inc. Method and apparatus for codon determining
US6961664B2 (en) 1999-01-19 2005-11-01 Maxygen Methods of populating data structures for use in evolutionary simulations
ES2341217T3 (es) 1999-01-19 2010-06-17 Maxygen, Inc. Recombinacion de acidos nucleicos mediada por oligonucleotidos.
AU3391900A (en) 1999-03-05 2000-09-21 Maxygen, Inc. Encryption of traits using split gene sequences
US7430477B2 (en) 1999-10-12 2008-09-30 Maxygen, Inc. Methods of populating data structures for use in evolutionary simulations
US6519065B1 (en) 1999-11-05 2003-02-11 Jds Fitel Inc. Chromatic dispersion compensation device
EP1272967A2 (en) 2000-03-30 2003-01-08 Maxygen, Inc. In silico cross-over site selection
US20050084907A1 (en) 2002-03-01 2005-04-21 Maxygen, Inc. Methods, systems, and software for identifying functional biomolecules
US7747391B2 (en) 2002-03-01 2010-06-29 Maxygen, Inc. Methods, systems, and software for identifying functional biomolecules
JP5319865B2 (ja) 2002-03-01 2013-10-16 コデクシス メイフラワー ホールディングス， エルエルシー 機能的生体分子を同定する方法、システム、およびソフトウェア
EP1488335A4 (en) 2002-03-09 2006-11-15 Maxygen Inc OPTIMIZING CROSSOVER POINTS FOR THE ADJUSTED EVOLUTION
EP1639091B1 (en) * 2003-06-17 2012-12-05 California University Of Technology Regio- and enantioselective alkane hydroxylation with modified cytochrome p450
GB0403992D0 (en) * 2004-02-23 2004-03-31 Isis Innovation Oxidation by hydrogen peroxide
CN1273585C (zh) * 2004-11-29 2006-09-06 浙江大学 细胞色素 p450bm－3 单加氧酶变体基因及其用途
JP2009514969A (ja) * 2005-11-09 2009-04-09 コンビナトアールエックス インコーポレーティッド 医学的状態を治療するための方法、組成物、およびキット
PT2068938E (pt) * 2006-09-22 2011-03-23 Novartis Ag Optimização do tratamento da leucemia filadélfia positiva com o inibidor de tirosina quinase abl imatinib
JP2010504978A (ja) * 2006-09-29 2010-02-18 アイデニクス ファーマシューティカルズ，インコーポレーテッド Ｈｉｖ阻害剤としての鏡像異性的に純粋なホスホインドール
US8101399B2 (en) 2006-12-12 2012-01-24 The Regents Of The University Of California Artemisinic epoxide and methods for producing same
US20080268517A1 (en) * 2007-02-08 2008-10-30 The California Institute Of Technology Stable, functional chimeric cytochrome p450 holoenzymes
US8802401B2 (en) * 2007-06-18 2014-08-12 The California Institute Of Technology Methods and compositions for preparation of selectively protected carbohydrates
GB0719620D0 (en) * 2007-10-08 2007-11-14 Isis Innovation Mutant Enzymes
US8383346B2 (en) 2008-06-13 2013-02-26 Codexis, Inc. Combined automated parallel synthesis of polynucleotide variants
DK3023494T3 (en) 2008-06-13 2019-02-25 Codexis Inc PROCEDURE FOR SYNTHESIS OF POLYNUCLEOTIDE VARIETIES
US20090312196A1 (en) 2008-06-13 2009-12-17 Codexis, Inc. Method of synthesizing polynucleotide variants
EP2426198A1 (en) * 2010-09-03 2012-03-07 B.R.A.I.N. Biotechnology Research And Information Network AG Cytochrome P450 monooxygenase variants
WO2012109586A2 (en) * 2011-02-11 2012-08-16 University Of Rochester Methods and systems for evaluating and predicting the reactivity of monooxygenase enzymes
WO2013076258A2 (en) * 2011-11-23 2013-05-30 Dsm Ip Assets B.V. P450 bm3 mutants and their use for regio- and stereoselective hydroxylation of alpha-and beta-ionone
CN102757944A (zh) * 2012-07-24 2012-10-31 浙江大学宁波理工学院 细胞色素p450bm-3(d168l/e435t)变体酶及其编码基因和用途
JP2015534464A (ja) * 2012-10-09 2015-12-03 カリフォルニア インスティチュート オブ テクノロジー ヘム酵素によって触媒されるインビボおよびインビトロオレフィンシクロプロパン化
US9708587B2 (en) * 2014-07-09 2017-07-18 Codexis, Inc. P450-BM3 variants with improved activity
US9683220B2 (en) * 2015-07-07 2017-06-20 Codexis, Inc. P450-BM3 variants with improved activity

Also Published As

Publication number Publication date
US20170247667A1 (en) 2017-08-31
CA2991195A1 (en) 2017-01-12
US10450550B2 (en) 2019-10-22
DK3319987T3 (da) 2021-06-21
US20190024059A1 (en) 2019-01-24
EP3319987A4 (en) 2018-12-19
JP2018520674A (ja) 2018-08-02
US20230250403A1 (en) 2023-08-10
US10113153B2 (en) 2018-10-30
EP3319987B1 (en) 2021-05-05
US10982197B2 (en) 2021-04-20
US20200002685A1 (en) 2020-01-02
WO2017007547A1 (en) 2017-01-12
US11591578B2 (en) 2023-02-28
US20200291365A1 (en) 2020-09-17
JP6691558B2 (ja) 2020-04-28
IL256441A (en) 2018-02-28
EP3319987A1 (en) 2018-05-16
IL256441B (en) 2020-02-27
CA2991195C (en) 2021-06-29
US20210214694A1 (en) 2021-07-15
CN108271374A (zh) 2018-07-10
US20170009213A1 (en) 2017-01-12
US9683220B2 (en) 2017-06-20
US10704030B2 (en) 2020-07-07

Similar Documents

Publication Publication Date Title
US12365882B2 (en) P450-BM3 variants with improved activity
US11591578B2 (en) P450-BM3 variants with improved activity
US10793841B2 (en) T7 RNA polymerase variants
WO2018208665A1 (en) Engineered ligase variants
US20170362616A1 (en) Biosynthetic production of caffeine
US12129495B2 (en) Engineered DNA polymerase variants

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant