JPH04500756A - 熱、酸及び/またはアルカリに高い安定性を有する細菌由来突然変異体α―アミラーゼ - Google Patents
熱、酸及び/またはアルカリに高い安定性を有する細菌由来突然変異体α―アミラーゼInfo
- Publication number
- JPH04500756A JPH04500756A JP2509959A JP50995990A JPH04500756A JP H04500756 A JPH04500756 A JP H04500756A JP 2509959 A JP2509959 A JP 2509959A JP 50995990 A JP50995990 A JP 50995990A JP H04500756 A JPH04500756 A JP H04500756A
- Authority
- JP
- Japan
- Prior art keywords
- amylase
- mutant
- starch
- gene
- 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.)
- Granted
Links
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
- 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
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
-
- 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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.)
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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
Abstract
Description
Claims (21)
- 1.野生型α−アミラーゼと少なくとも1個のアミノ酸が異なるアミノ酸配列を 持ち、さらにでん粉の分解、及び/あるいは繊維の糊抜への応用においてそのア ミノ酸置換により性質が向上したことを特徴とする、α−アミラーゼをコードす るDNA配列に変異をおこしたDNAの発現産物であ変異体α−アミラーゼ。
- 2.高い熱安定性を示すことを特徴とする請求の範囲第1項記載のα−アミラー ゼ。
- 3.pH6.5以下かつ/あるいは7.5以上で高い安定性を示すことを特徴と する、請求の範囲第一項記載のα−アミラーゼ。
- 4.高い熱安定性及び酸安定性を示すことを特徴とする、請求の範囲第12項記 載のα−アミラーゼ。
- 5.変異体酵素を構築する元にした遺伝子が微生物;好ましくは枯草菌から得ら れる請求の範囲第1項から第4項のいずれか1項に記載のα−アミラーゼ。
- 6.該遺伝子がバチルスステアロテルモフィルス、バチルスリケニホルミス、バ チラスアミロリクエファシエンスからなる群まから選択した株の野生型遺伝子を 元に構築した該遺伝子による請求の範囲第5項記載のα−アミラーゼ。
- 7.バチラスリケニホルミスから得られる野生型α−アミラーゼと111、13 3、149番目のうちの1つ以上、あるいは相同のα−アミラーゼの相当する位 置において1個のアミノ酸置換により異なることを特徴とする請求の範囲第6項 記載のα−アミラーゼ。
- 8.次のアミノ配置換、すなわちAla−111−Thr,His−133−T yr,Thr−149−Ileの1つ以上を含むことを特徴とする、請求の範囲 第7項記載のα−アミラーゼ。
- 9.請求の範囲第1項から第8項中の1つに定義されたα−アミラーゼをコード する変異体遺伝子
- 10.請求の範囲第9項記載の変異体遺伝子を含む発現ベクター。
- 11.請求の範囲第10項記載の発現ベクターを含む宿主細胞。
- 12.請求の範囲第10項記載の発現ベクターにより形質転換されることを特徴 とし、形質転換に先立ち、細胞外澱粉分解酵素を産生する能力が実質上ない宿主 細胞。
- 13.バチルスリケニホルミスT9を表した請求の範囲第12項記載の宿主細胞 。
- 14.構造遺伝子から調節配列を物理的に分離することにより大腸菌内でクロー ン化した遺伝子の発現を不可能にし、また枯草菌中でクローン化した遺伝子の発 現を1種の制限酵素による消化及びその後の再環状化により回復することのでき る枯草菌/大鵬菌シャトルベクター。
- 15.澱粉分解または繊維糊抜きへの応用に際し向上した性質をもつ澱粉分解酵 素を調製する方法であって以下の工程;澱粉分解酵素をコードするクローン化し た遺伝子あるいはその遺伝子断片に突然変異をおこす工程;得られた変異体アミ ラーゼ遺伝子あるいは遺伝子群を単離する工程;該変異体アミラーゼ遺伝子(群 )を発現する工程;産生に適当な宿主細胞株に導入する工程;及び産生した変異 体アミラーゼを回収し澱粉分解あるいは繊維糊抜きへの応用において向上した性 質をもつ変異体アミラーゼを同定する工程を含む方法。
- 16.変異体α−アミラーゼを産生する方法であって、請求の範囲第11項から 第13項のうちの1つに記載の宿主細胞を適当な培地中で培養する工程、及び産 生されたα−アミラーゼを回収する工程を含む方法。
- 17.澱粉分解及び繊維糊抜きでの請求の範囲第1項から第8項記載のα−アミ ラーゼの利用。
- 18.請求の範囲第1項から第8項のうちの1つに記載した変異体α−アミラー ゼの利用を含む澱粉分解の方法。
- 19.請求の範囲第1項から第8項のうちの1つに記載した変異体α−アミラー ゼの利用を含む繊維糊抜き方法。
- 20.請求の範囲第1項から第8項のうちの1つに記載した変異体α−アミラー ゼを含む澱粉分解用組成物。
- 21.請求の範囲第1項から第8項のうちの1つに記載した変異体α−アミラー ゼを含む繊維糊抜き用組成物。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP89201735 | 1989-06-29 | ||
EP89201735.1 | 1989-06-29 | ||
PCT/EP1990/001042 WO1991000353A2 (en) | 1989-06-29 | 1990-06-27 | MUTANT MICROBIAL α-AMYLASES WITH INCREASED THERMAL, ACID AND/OR ALKALINE STABILITY |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP11305433A Division JP2000197491A (ja) | 1989-06-29 | 1999-10-27 | 枯草菌/大腸菌シャトルベクター |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH04500756A true JPH04500756A (ja) | 1992-02-13 |
JP3086249B2 JP3086249B2 (ja) | 2000-09-11 |
Family
ID=8202426
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP02509959A Expired - Lifetime JP3086249B2 (ja) | 1989-06-29 | 1990-06-27 | 熱、酸及び/またはアルカリに高い安定性を有する細菌由来突然変異体α―アミラーゼ |
JP11305433A Pending JP2000197491A (ja) | 1989-06-29 | 1999-10-27 | 枯草菌/大腸菌シャトルベクター |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP11305433A Pending JP2000197491A (ja) | 1989-06-29 | 1999-10-27 | 枯草菌/大腸菌シャトルベクター |
Country Status (17)
Country | Link |
---|---|
US (1) | US5364782A (ja) |
EP (1) | EP0410498B1 (ja) |
JP (2) | JP3086249B2 (ja) |
KR (1) | KR0165550B1 (ja) |
CN (1) | CN1050220A (ja) |
AT (1) | ATE166922T1 (ja) |
BG (1) | BG61081B1 (ja) |
BR (1) | BR9006818A (ja) |
CA (1) | CA2030554C (ja) |
DD (1) | DD301620A9 (ja) |
DE (1) | DE69032360T2 (ja) |
DK (1) | DK0410498T3 (ja) |
ES (1) | ES2117625T3 (ja) |
FI (1) | FI103285B1 (ja) |
IE (1) | IE902369A1 (ja) |
PT (1) | PT94560B (ja) |
WO (1) | WO1991000353A2 (ja) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994026881A1 (en) * | 1993-05-19 | 1994-11-24 | Kao Corporation | LIQUEFYING ALKALINE α-AMYLASE, PROCESS FOR PRODUCING THE SAME, AND DETERGENT COMPOSITION CONTAINING THE SAME |
JP2009072204A (ja) * | 1996-04-30 | 2009-04-09 | Novozyme As | α−アミラーゼ変異体 |
JP2011125341A (ja) * | 2003-04-01 | 2011-06-30 | Genencor Internatl Inc | 変異体フミコーラ・グリセアcbh1.1 |
JP2011525350A (ja) * | 2008-06-06 | 2011-09-22 | ダニスコ・ユーエス・インク | バシラス・スブチリス由来のアルファ−アミラーゼ変異体とその使用方法 |
Families Citing this family (128)
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WO1992019744A1 (en) * | 1991-05-06 | 1992-11-12 | Valio, Finnish Co-Operative Dairies' Association | ACID α-AMYLASE |
JPH07503363A (ja) * | 1991-11-14 | 1995-04-13 | ノボザイムス アクティーゼルスカブ | バシラス・リヘニフォルミス中で遺伝子を発現させる方法 |
AU5659094A (en) * | 1992-12-09 | 1994-07-04 | Yoshiyuki Takasaki | Novel acid- and heat-resistant alpha-amylase, process for producing the same, and method of liquefying starch by using the same |
DK154292D0 (da) * | 1992-12-23 | 1992-12-23 | Novo Nordisk As | Nyt enzym |
DK0867504T4 (da) * | 1993-02-11 | 2011-08-29 | Genencor Int | Oxidativ stabil alfa-amylase |
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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 |
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- 1990-06-27 EP EP90201706A patent/EP0410498B1/en not_active Expired - Lifetime
- 1990-06-27 KR KR1019910700227A patent/KR0165550B1/ko not_active IP Right Cessation
- 1990-06-27 JP JP02509959A patent/JP3086249B2/ja not_active Expired - Lifetime
- 1990-06-27 AT AT90201706T patent/ATE166922T1/de not_active IP Right Cessation
- 1990-06-27 DE DE69032360T patent/DE69032360T2/de not_active Expired - Lifetime
- 1990-06-27 US US07/623,953 patent/US5364782A/en not_active Expired - Lifetime
- 1990-06-27 DK DK90201706T patent/DK0410498T3/da active
- 1990-06-27 WO PCT/EP1990/001042 patent/WO1991000353A2/en active IP Right Grant
- 1990-06-27 ES ES90201706T patent/ES2117625T3/es not_active Expired - Lifetime
- 1990-06-27 BR BR909006818A patent/BR9006818A/pt not_active Application Discontinuation
- 1990-06-27 CA CA002030554A patent/CA2030554C/en not_active Expired - Lifetime
- 1990-06-29 PT PT94560A patent/PT94560B/pt not_active IP Right Cessation
- 1990-06-29 CN CN90106811A patent/CN1050220A/zh active Pending
- 1990-06-29 DD DD90342352A patent/DD301620A9/de unknown
- 1990-06-29 IE IE236990A patent/IE902369A1/en unknown
-
1991
- 1991-02-11 BG BG93814A patent/BG61081B1/bg unknown
- 1991-02-25 FI FI910907A patent/FI103285B1/fi active
-
1999
- 1999-10-27 JP JP11305433A patent/JP2000197491A/ja active Pending
Cited By (6)
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WO1994026881A1 (en) * | 1993-05-19 | 1994-11-24 | Kao Corporation | LIQUEFYING ALKALINE α-AMYLASE, PROCESS FOR PRODUCING THE SAME, AND DETERGENT COMPOSITION CONTAINING THE SAME |
CN1062906C (zh) * | 1993-05-19 | 2001-03-07 | 花王株式会社 | 液化碱性α-淀粉酶、其制备方法及含有它的洗涤剂组合物 |
JP2009072204A (ja) * | 1996-04-30 | 2009-04-09 | Novozyme As | α−アミラーゼ変異体 |
JP4580012B2 (ja) * | 1996-04-30 | 2010-11-10 | ノボザイムス アクティーゼルスカブ | α−アミラーゼ変異体 |
JP2011125341A (ja) * | 2003-04-01 | 2011-06-30 | Genencor Internatl Inc | 変異体フミコーラ・グリセアcbh1.1 |
JP2011525350A (ja) * | 2008-06-06 | 2011-09-22 | ダニスコ・ユーエス・インク | バシラス・スブチリス由来のアルファ−アミラーゼ変異体とその使用方法 |
Also Published As
Publication number | Publication date |
---|---|
DK0410498T3 (da) | 1999-03-22 |
IE902369A1 (en) | 1991-06-19 |
JP2000197491A (ja) | 2000-07-18 |
KR0165550B1 (ko) | 1999-01-15 |
US5364782A (en) | 1994-11-15 |
EP0410498A2 (en) | 1991-01-30 |
CA2030554C (en) | 2001-08-28 |
AU638263B2 (en) | 1993-06-24 |
FI103285B (fi) | 1999-05-31 |
WO1991000353A2 (en) | 1991-01-10 |
KR920701449A (ko) | 1992-08-11 |
ES2117625T3 (es) | 1998-08-16 |
BG61081B1 (bg) | 1996-10-31 |
EP0410498A3 (en) | 1991-11-06 |
PT94560B (pt) | 1998-01-30 |
JP3086249B2 (ja) | 2000-09-11 |
PT94560A (pt) | 1991-02-08 |
AU5953890A (en) | 1991-01-17 |
WO1991000353A3 (en) | 1991-02-21 |
CA2030554A1 (en) | 1990-12-30 |
CN1050220A (zh) | 1991-03-27 |
DE69032360D1 (de) | 1998-07-09 |
DE69032360T2 (de) | 1998-12-03 |
EP0410498B1 (en) | 1998-06-03 |
IE902369L (en) | 1990-12-29 |
FI910907A0 (fi) | 1991-02-25 |
ATE166922T1 (de) | 1998-06-15 |
BR9006818A (pt) | 1991-08-06 |
DD301620A9 (de) | 1993-04-29 |
FI103285B1 (fi) | 1999-05-31 |
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