JP2001506496A - 改良された安定性を有するタンパク質 - Google Patents
改良された安定性を有するタンパク質Info
- Publication number
- JP2001506496A JP2001506496A JP52659698A JP52659698A JP2001506496A JP 2001506496 A JP2001506496 A JP 2001506496A JP 52659698 A JP52659698 A JP 52659698A JP 52659698 A JP52659698 A JP 52659698A JP 2001506496 A JP2001506496 A JP 2001506496A
- Authority
- JP
- Japan
- Prior art keywords
- protein
- amino acid
- amylase
- bacillus
- enzyme
- 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.)
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Classifications
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
- C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38627—Preparations containing enzymes, e.g. protease or amylase containing lipase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38636—Preparations containing enzymes, e.g. protease or amylase containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38654—Preparations containing enzymes, e.g. protease or amylase containing oxidase or reductase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Animal Husbandry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Cosmetics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.低安定性であるが相同性のタンパク質中の対応するアミノ酸残基とは異なる アミノ酸残基の置換又は欠失によって、前駆体アミノ酸配列から修飾されたアミ ノ酸配列を含んでなる改良されたタンパク質であって、前記改良されたタンパク 質が前駆体アミノ酸配列に対応するタンパク質と比較して安定性が変化している 改良されたタンパク質。 2.前記タンパク質が酵素を包含する請求の範囲第1項に記載のタンパク質。 3.前記酵素が、加水分解酵素、酸化還元酵素、転移酵素、リアーゼ又はリガー ゼを包含する請求の範囲第2項に記載の酵素。 4.前記酵素が、アミラーゼ、リパーゼ、セルラーゼ、プロテアーゼ、ヘミセル ラーゼ、グルコアミラーゼ、エステラーゼ、ラクターゼ、ポリガラクトロナーゼ 、β-ガラクトシダーゼ又はリグニナーゼを包含する請求の範囲第3項に記載の 酵素。 5.前記前駆体アミノ酸配列が、バシラス リケニホルミス由来のα-アミラー ゼに対応するアミノ酸配列を包含し、前記低安定性であるが相同性の酵素が、バ シラス ステアロサーモフィラス、バシラス サブチリス又はバシラス アミロリ クエファシエンスを包含する請求の範囲第4項に記載のα-アミラーゼ。 6.前記置換されたアミノ酸残基が、バシラス ステアロサーモフィラスとバシ ラス アミロリクエファシエンスの両方にある対応する残基とは異なるアミノ酸 残基から選択され、かつ、前記バシラス ステアロサーモフィラスとバシラス ア ミロリクエファシエンスの両方にある対応する残基が同一のアミノ酸である請求 の範囲第5項に記載のα-アミラーゼ。 7.前記置換されたアミノ酸残基が、バシラス ステアロサーモフィラスとバシ ラス アミロリクエファシエンスの両方にある対応する残基に存在する同じアミ ノ酸で置換されている請求の範囲第6項に記載のα-アミラーゼ。 8.前記置換されたアミノ酸残基が、A33、A52、S85、N96、H133、S148、A209、 A269、A379及びA435に対応する請求の範囲第5項に記載のα-アミラーゼ。 9.前記置換されたアミノ酸残基が、A33S、A52S、N96Q、H133Y、S148N、 A209V、A269K、A379S及び/又はA435Sに対応する請求の範囲第7項に記載のα- アミラーゼ。 10.さらにM15、N188及び/又はM197における置換を包含する請求の範囲第7項 に記載のα-アミラーゼ。 11.前記置換が、M15T/A33S/H133Y/N188S/A209V;M15T/D28N/A33S/H133Y /N188S/A209V;M15T/A52S/H133Y/N188S/A209V;M15T/A52S/H133Y/N188 S/A209V/L230F;M15T/S85D/H133Y/N188S/A209V;M15T/N96Q/H133Y/N18 8S/A209V;M15T/N96Q/H133Y/N188S/A209V/I479T;M15T/H133Y/S148N/N 188S/A209V;M15T/H133Y/S148N/N188S/A209V/A379S;M15T/H133Y/S148N /N188S/A209V/G433D;M15T/H133Y/N188S/A209V/A379S;M15T/H133Y/N1 88S/A209V/A210S/T322A/A379Sを包含する請求の範囲第10項に記載のα-アミ ラーゼ。 12.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも60%の配列同一性を有する請求の範囲第1項に記載のタンパク質。 13.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも65%の配列同一性を有する請求の範囲第12項に記載のタンパク質。 14.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも80%の配列同一性を有する請求の範囲第12項に記載のタンパク質。 15.前記修飾されたタンパク質が、前記前駆体タンパク質と比較して改良された 安定性を有する請求の範囲第1項に記載のタンパク質。 16.(a)低安定性であるが相同性の酵素を有する前駆体タンパク質の配列を調整 し、その前駆体タンパク質及び前記低安定性であるが相同性の酵素間で異なる1 以上の残基を同定する工程; (b)前記前駆体タンパク質中の置換のために、工程(a)で同定された残基を1以上 選択する工程; (c)前記前駆体タンパク質を修飾して、工程(b)で選択された置換用残基を導入す る工程 を包含する、 前駆体タンパク質に基づく改良されたタンパク質の製造方法。 17.前記タンパク質が、酵素を包含する請求の範囲第16項に記載の方法。 18.前記酵素が、加水分解酵素、酸化還元酵素、転移酵素、リアーゼ又はリガー ゼを包含する請求の範囲第17項に記載の方法。 19.前記酵素が、アミラーゼ、リパーゼ、セルラーゼ、プロテアーゼ、ヘミセル ラーゼ、グルコアミラーゼ、エステラーゼ、ラクターゼ、ポリガラクトロナーゼ 、β-ガラクトシダーゼ又はリグニナーゼを包含する請求の範囲第17項に記載の 方法。 20.前記前駆体アミノ酸配列が、バシラス リケニホルミス由来のα-アミラーゼ に対応するアミノ酸配列を包含し、前記低安定性であるが相同性の酵素が、バシ ラス ステアロサーモフィラス、バシラス サブチリス又はバシラス アミロリク エファシエンスを包含する請求の範囲第19項に記載の方法。 21.前記置換されたアミノ酸残基が、バシラス ステアロサーモフィラスとバシ ラス アミロリクエファシエンスの両方にある対応する残基とは異なるアミノ酸 残基から選択され、かつ、前記バシラス ステアロサーモフィラスとバシラス ア ミロリクエファシエンスの両方にある対応する残基が同一のアミノ酸である請求 の範囲第20項に記載の方法。 22.前記置換されたアミノ酸残基が、バシラス ステアロサーモフィラスとバシ ラス アミロリクエファシエンスの両方にある対応する残基に存在する同じアミ ノ酸で置換されている請求の範囲第21項に記載の方法。 23.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも60%の配列同一性を有する請求の範囲第16項に記載の方法。 24.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも65%の配列同一性を有する請求の範囲第12項に記載の方法。 25.前記低安定性であるが相同性のタンパク質が、前記前駆体タンパク質と少な くとも80%の配列同一性を有する請求の範囲第12項に記載の方法。 26.(a) デンプン水溶液を調製する工程;及び (b) 前記デンプン水溶液を、請求の範囲第5項に記載のα-アミラーゼと接 触させる工程 を包含するデンプンの液化方法。 27.請求の範囲第2項に記載の酵素を含有する洗浄剤組成物。 28.請求の範囲第2項に記載の酵素を含有する食品添加剤組成物。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/US1996/019595 WO1998026078A1 (en) | 1996-12-09 | 1996-12-09 | H mutant alpha-amylase enzymes |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2001506496A true JP2001506496A (ja) | 2001-05-22 |
JP4253041B2 JP4253041B2 (ja) | 2009-04-08 |
Family
ID=22256262
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP52659698A Expired - Lifetime JP4253041B2 (ja) | 1996-12-09 | 1996-12-09 | 改良された安定性を有するタンパク質 |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP0942994B1 (ja) |
JP (1) | JP4253041B2 (ja) |
AT (1) | ATE293696T1 (ja) |
AU (1) | AU1461497A (ja) |
CA (1) | CA2274806C (ja) |
DE (1) | DE69634640T2 (ja) |
WO (1) | WO1998026078A1 (ja) |
Families Citing this family (82)
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US6440716B1 (en) | 1995-02-03 | 2002-08-27 | Novozymes A/S | α-amylase mutants |
US6080568A (en) * | 1997-08-19 | 2000-06-27 | Genencor International, Inc. | Mutant α-amylase comprising modification at residues corresponding to A210, H405 and/or T412 in Bacillus licheniformis |
US6887986B1 (en) | 1998-11-16 | 2005-05-03 | Novozymes A/S | α-amylase variants |
US6197565B1 (en) * | 1998-11-16 | 2001-03-06 | Novo-Nordisk A/S | α-Amylase variants |
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US6410295B1 (en) | 1999-03-30 | 2002-06-25 | Novozymes A/S | Alpha-amylase variants |
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JP4658433B2 (ja) * | 1999-12-23 | 2011-03-23 | ジェネンコア インターナショナル インコーポレーテッド | 改良された機能特性を有するタンパク質を得る方法 |
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BR9407767A (pt) * | 1993-10-08 | 1997-03-18 | Novo Nordisk As | Variante de enzima &-amilase uso da mesma construção de DNA vetor de express o recombinante célula processos para produzir uma &-amilase hibrida e para preparar uma variante de uma &-amilase aditivo detergente e composições detergentes |
AU685638B2 (en) * | 1994-06-17 | 1998-01-22 | Genencor International, Inc. | Novel amylolytic enzymes derived from the b. licheniformis alpha-amylase, having improved characteristics |
ES2329528T3 (es) * | 1995-02-03 | 2009-11-26 | Novozymes A/S | Metodo para diseñar mutantes alfa-amilasa con propiedades determinadas. |
US5736499A (en) * | 1995-06-06 | 1998-04-07 | Genencor International, Inc. | Mutant A-amylase |
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1996
- 1996-12-09 WO PCT/US1996/019595 patent/WO1998026078A1/en active IP Right Grant
- 1996-12-09 CA CA2274806A patent/CA2274806C/en not_active Expired - Lifetime
- 1996-12-09 AT AT96945189T patent/ATE293696T1/de not_active IP Right Cessation
- 1996-12-09 EP EP96945189A patent/EP0942994B1/en not_active Expired - Lifetime
- 1996-12-09 DE DE69634640T patent/DE69634640T2/de not_active Expired - Lifetime
- 1996-12-09 AU AU14614/97A patent/AU1461497A/en not_active Abandoned
- 1996-12-09 JP JP52659698A patent/JP4253041B2/ja not_active Expired - Lifetime
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EP0942994A1 (en) | 1999-09-22 |
DE69634640D1 (de) | 2005-05-25 |
EP0942994B1 (en) | 2005-04-20 |
CA2274806C (en) | 2011-02-01 |
DE69634640T2 (de) | 2006-01-19 |
ATE293696T1 (de) | 2005-05-15 |
WO1998026078A1 (en) | 1998-06-18 |
JP4253041B2 (ja) | 2009-04-08 |
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