JPH10510167A - ポリケチドの無細胞合成 - Google Patents
ポリケチドの無細胞合成Info
- Publication number
- JPH10510167A JPH10510167A JP9505304A JP50530497A JPH10510167A JP H10510167 A JPH10510167 A JP H10510167A JP 9505304 A JP9505304 A JP 9505304A JP 50530497 A JP50530497 A JP 50530497A JP H10510167 A JPH10510167 A JP H10510167A
- Authority
- JP
- Japan
- Prior art keywords
- pks
- polyketide
- unit
- cell
- free
- 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.)
- Granted
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/08—Hetero rings containing eight or more ring members, e.g. erythromycins
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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
- 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
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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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
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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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/08—Oxygen as only ring hetero atoms containing a hetero ring of at least seven ring members, e.g. zearalenone, macrolide aglycons
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Indole Compounds (AREA)
- Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.所望のポリケチドを生成する方法であって、 (a)無細胞系内にモジュラーポリケチドシンターゼの少なくとも2つのモジ ュールを含む1つまたはそれ以上のタンパク質を提供する工程と、 (b)少なくとも1つの開始ユニットおよび少なくとも1つの延長ユニットを 該系に添加する工程と、 (c)該開始ユニットおよび該延長ユニットを含む該無細胞系を、該ポリケチ ドが合成される条件下でインキュベートする工程と、 (d)必要に応じて該ポリケチドを該無細胞系から回収する工程と、 を包含する方法。 2.前記開始ユニットおよび前記延長ユニットが異なる、請求項1に記載の方法 。 3.前記ポリケチドの合成条件がNADPHの添加を含む、請求項1に記載の方法。 4.工程(a)において、モジュラーPKSの少なくとも3モジュールを含む少な くとも2つのタンパク質が提供される、請求項1に記載の方法。 5.前記1つのタンパク質または複数のタンパク質が、少なくとも2つの異なる モジュラーPKSからのモジュールを含む、請求項1に記載の方法。 6.所望のポリケチドを生成する方法であって、 (a)機能性モジュラーポリケチドシンターゼ(PKS)、またはその機能性部 分を提供する工程であって、ここで、該PKSには、天然の第1モジュール開始ユ ニットがロードされ得ず、またはここで、一旦ロードされると、該PKSは、延長 ユニットの該第1モジュール開始ユニットへの縮合を触媒し得ず、ポリケチド中 間体を生成し得ない、工程と、 (b)該PKSの基質である開始ユニットを該系に添加する工程と、 (c)該ポリケチドが合成される条件下で、該PKSおよび該開始ユニット基質 を含む該系をインキュベートする工程と、 (d)必要に応じて該ポリケチドを回収する工程と、 を包含する方法。 7.ポリケチドのライブラリーを調製する方法であって、 (a)多数の無細胞系またはサブ系を提供する工程であって、各系またはサブ 系は、少なくとも1つの延長ユニットのポリケチド鎖へのカップリングを達成す る酵素活性を含む1つまたはそれ以上のポリケチドシンターゼタンパク質ならび に少なくとも1つの開始ユニットおよび少なくとも1つの延長ユニットを含む、 工程と、 (b)該開始ユニットおよび該延長ユニットを含む該無細胞系またはサブ系を 、該ポリケチドが合成される条件下でインキュベートする工程と、 (c)必要に応じて該ポリケチドを該無細胞系またはサブ系から回収する工程 と、 を包含する方法。 8.少なくとも1つの系またはサブ系内の前記開始ユニットおよび前記延長ユニ ットが、互いに異なる、請求項7に記載の方法。 9.前記無細胞系のそれぞれが少なくとも2つのサブ系に細分され、該サブ系の それぞれが、少なくとも1つの開始ユニットおよび少なくとも1つの延長ユニッ トを含み、ここで、該開始ユニットおよび該延長ユニットの少なくとも1つが、 該同一の無細胞系の該サブ系のそれぞれの間で異なる、請求項7に記載の方法。 10.少なくとも2つの異なる無細胞系またはサブ系が、同一の開始ユニットお よび同一の延長ユニットを含む、請求項7に記載の方法。 11.無細胞サブ系、基質開始ユニットおよび基質延長ユニットのマトリックス であって、 (a)それぞれが、少なくとも1つの延長ユニットとポリケチド鎖との結合に 影響を与える酵素活性を含む1つまたはそれ以上のポリケチドシンターゼタンパ ク質を含む一連の無細胞サブ系を含み、 (b)該サブ系のそれぞれは、少なくとも1つの開始ユニットおよび少なくと も1つの延長ユニットを含み、そして (c)少なくとも1つの酵素活性または少なくとも1つの延長ユニットまたは 少なくとも1つの開始ユニットが該遺伝子内の各サブ系間で異なる、 マトリックス。 12.前記遺伝子内の前記サブ系が、2次元アレイ内に配置され、ここで、1つ の次元において、各サブ系は、少なくとも1つの任意の他のサブ系とは異なる酵 素活性を有し、同一の開始ユニットおよび/または延長ユニットを有し、他の次 元において、各サブ系は、他のサブ系と同一のポリケチドシンターゼ酵素活性を 含み、そして少なくとも1つの異なる開始サブユニットまたは延長サブユニット を有する、請求項11に記載のマトリックス。 13.前記ポリケチドシンターゼタンパク質が、モジュラーポリケチドシンター ゼ由来の酵素活性を含む、請求項11に記載のマトリックス。 14.前記ポリケチドシンターゼタンパク質が、芳香族ポリケチドシンターゼ由 来の酵素活性を含む、請求項11に記載のマトリックス。 15.前記ポリケチドシンターゼタンパク質が、モジュラーPKSおよび芳香族PKS の両方由来の酵素活性を含む、請求項11に記載のマトリックス。 16.天然の第1モジュール開始ユニットでロードされ得ず、または一旦ロード されると、延長ユニットの該第1モジュール開始ユニットへの縮合を触媒し得ず 、ポリケチド中間体を生成し得ない、機能性モジュラーポリケチドシンターゼ系 ま たはその機能性部分。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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US333895P | 1995-07-06 | 1995-07-06 | |
US60/003,338 | 1995-07-06 | ||
PCT/US1996/011317 WO1997002358A1 (en) | 1995-07-06 | 1996-07-05 | Cell-free synthesis of polyketides |
Publications (2)
Publication Number | Publication Date |
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JPH10510167A true JPH10510167A (ja) | 1998-10-06 |
JP3633629B2 JP3633629B2 (ja) | 2005-03-30 |
Family
ID=21705358
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP50530497A Expired - Fee Related JP3633629B2 (ja) | 1995-07-06 | 1996-07-05 | ポリケチドの無細胞合成 |
Country Status (10)
Country | Link |
---|---|
US (5) | US6080555A (ja) |
EP (2) | EP1493814A1 (ja) |
JP (1) | JP3633629B2 (ja) |
AT (1) | ATE286133T1 (ja) |
AU (1) | AU706445B2 (ja) |
CA (1) | CA2226221C (ja) |
DE (1) | DE69634127D1 (ja) |
IL (2) | IL122859A0 (ja) |
NZ (1) | NZ313383A (ja) |
WO (1) | WO1997002358A1 (ja) |
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- 1996-07-05 WO PCT/US1996/011317 patent/WO1997002358A1/en active IP Right Grant
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EP0836649A1 (en) | 1998-04-22 |
EP0836649B1 (en) | 2004-12-29 |
US6080555A (en) | 2000-06-27 |
CA2226221A1 (en) | 1997-01-23 |
EP1493814A1 (en) | 2005-01-05 |
DE69634127D1 (de) | 2005-02-03 |
US6750040B1 (en) | 2004-06-15 |
US20040203119A1 (en) | 2004-10-14 |
WO1997002358A1 (en) | 1997-01-23 |
AU6542696A (en) | 1997-02-05 |
EP0836649A4 (en) | 2000-04-05 |
US6531299B1 (en) | 2003-03-11 |
US7101684B2 (en) | 2006-09-05 |
AU706445B2 (en) | 1999-06-17 |
IL122859A (en) | 2009-05-04 |
NZ313383A (en) | 1999-08-30 |
CA2226221C (en) | 2001-05-08 |
JP3633629B2 (ja) | 2005-03-30 |
ATE286133T1 (de) | 2005-01-15 |
IL122859A0 (en) | 1998-08-16 |
US6274560B1 (en) | 2001-08-14 |
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