FI122574B - Oxidationsbeständig alfaamylas - Google Patents

Oxidationsbeständig alfaamylas Download PDF

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Publication number
FI122574B
FI122574B FI20080005A FI20080005A FI122574B FI 122574 B FI122574 B FI 122574B FI 20080005 A FI20080005 A FI 20080005A FI 20080005 A FI20080005 A FI 20080005A FI 122574 B FI122574 B FI 122574B
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FI
Finland
Prior art keywords
amylase
alpha
gly
mutated
leu
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FI20080005A
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English (en)
Finnish (fi)
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FI20080005A (sv
Inventor
Richard L Antrim
Christopher C Barnett
Colin Mitchinson
Scott D Power
Leif P Solheim
Carol A Requadt
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Genencor Int
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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
    • 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
    • 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
    • C12N15/74—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
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase

Claims (24)

1. Mutant alfa-amylas valt ur gruppen bestäende av: 5 a) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas frän Bacillus licheniformis genom punktspeci-fik substitution av en aminosyra i position M+15, och 10 b) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas, som är ett Bacillus-alfa-amylas, genom punkt-specifik substitution av en metioningrupp vars position 15 antingen i primär- eller tertiärstrukturen motsvarar M+15 i Bacillus licheniformis alfa-amylas, varvid nämnda alfa-amylas uppvisar en förändrad pH- och/eller temperaturverkningsprofil i jämförelse med alfa-amylas hos Bacillus av vildtyp; varvid den ersättande 20 aminosyran är Vai.
2. Mutant alfa-amylas enligt patentkrav 1, vilket dessutom innehäller en eller flera andra punktspecifika mutationer.
3. Mutant alfa-amylas enligt nägot av patentkraven 25. eller 2, väri prekursorn är Bacillus licheniformis alfa- amylas . CVJ
4. DNA som kodar ett muterat alfa-amylas enligt CM ^ nägot av patentkraven 1-3.
° 5. Expressionsvektorer, vilka kodar DNA:t enligt co ^ 30 patentkrav 4. x
£ 6. Värdceller, vilka transformerats med express- lo ionsvektorn enligt patentkrav 5. o
§ 7. Tvättmedelskomposition, vilken innehäller ett oo o muterat alfa-amylas enligt nägot av patentkraven 1-3. ^ 35
8. Tvättmedelskomposition enligt patentkrav 7, vilket är en vätske-, gel-, eller granulatkomposition.
9. Tvättmedelskomposition enligt patentkrav 7 el-ler 8, vilken dessutom innehäller en eller flera ytterli-gare enzym.
10. Stärkelsekondenserande komposition, vilken in-5 nehäller ett muterat alfa-amylas enligt nägot av patent- kraven 1 - 3.
11. Förfarande för att ändra pH och/eller tempera-turverkningsprofilen hos ett Bacillus-alfa-amylas, vilket omfattar steg där man: 10 a) skaffar en prekursor-DNA-sekvens som kodar ett prekursor-alfa-amylas av Bacillus licheniformis, och modi-fierar nämnda prekursor-DNA-sekvens för att erhälla en mu-terad DNA-sekvens som kodar ett muterat alfa-amylas, som skiljer sig frän nämnda prekursor-alfa-amylas genom punkt- 15 specifik substitution av en aminosyra i position M+15, eller b) skaffar en prekursor-DNA-sekvens som kodar ett prekursor-alfa-amylas frän Bacillus, och modifierar nämnda prekursor DNA-sekvens för att erhälla en muterad DNA-se- 20 kvens som kodar ett muterat alfa-amylas som skiljer sig frän nämnda prekursor-alfa-amylas genom punktspecifik substitution av en metioningrupp vars position antingen i primär- eller tertiärstrukturen motsvarar M+15 in Bacillus licheniformis alfa-amylas; 25 och uttrycker det muterade alfa-amylaset av nämnda muterade DNA-sekvens, varvid nämnda muterade alfa-amylas CM q uppvisar en ändrad pH och/eller temperaturverkningsprofil CM ^ jämfört med det Bacillus prekursor-alfa-amylaset, varvid den ersättande aminosyran är Thr. co 30
12. Förfarande enligt patentkrav 11, vilket ytter- ir ligare omfattar att man modifierar nämnda prekursor-DNA- ^ sekvens sä att det muterade alfa-amylaset innehäller en o o eller flera andra punktspecifikä mutationer. o
13. Förfarande enligt patentkrav 11 eller 12 där O ^ 35 prekursorn är Bacillus licheniformis alfa-amylas.
14. Förfarande enligt nägot av patentkraven 11 - 13, vilket ytterligare omfattar att man formulerar det mute-rade alfa-amylaset i en tvättmedelskomposition.
15. Förfarande enligt patentkrav 14, där tvättme-5 delskompositionen är en vätske-, gel- eller granulatkompo- sition.
16. Förfarande enligt patentkrav 14 eller 15, där tvättmedelskompositionen dessutom innehäller ett eller flera ytterligare enzym. 10
17. Förfarande enligt nägot av patentkraven 11 - 13, vilket ytterligare omfattar att man formulerar det mute-rade alfa-amylaset tili en komposition, som överför stär-kelse i flytande form.
18. Tvättmedelskomposition, vilken innehäller att 15 muterat alfa-amylas och en eller flera ytterligare enzym, varvid det muterade alfa-amylaset väljs ur gruppen bestä-ende av: a) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid 20 den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas frän Bacillus licheniformis genom punktspeci-fik substitution av en aminosyra i position M+15, och b) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid 25 den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas, som är ett Bacillus-alfa-amylas, genom punkt-5 specifik substitution av en metioningrupp vars position (M ^ antingen i primär- eller tertiärstrukturen motsvarar M+15 ° i Bacillus licheniformis alfa-amylas, CO ^ 30 varvid nämnda alfa-amylas uppvisar en förändrad X £ pH- och/eller temperaturverkningsprofil i jämförelse med LO alfa-amylas hos Bacillus av vildtyp; varvid den ersättande o g aminosyran är Leu, Thr, Asn, Asp, Ser, Vai eller Ile. oo
19. Tvättmedelskomposition enligt patentkrav 18, 0X1 35 där nämnda muterade alfa-amylas är M15L.
20. Tvättmedelskomposition enligt patentkrav 18 eller 19, där nämnda muterade alfa-amylas innehäller en eller flera övriga punktspecifikä mutationer.
21. Tvättmedelskomposition enligt nägot av patent-5 kraven 18 - 20, där nämnda ytterligare enzym eller enzymer väljs ur gruppen bestäende av amylas, proteaser, lipaser, och cellulaser.
22. Förfarande för att överföra granulatformigt stärkelseslam frän antingen en vat eller torr valsnings- 10 process i flytande form vid ett pH frän ca 4 tili ca 6, vilket omfattar att man: a) tillsätter en effektiv mängd muterat alfa-amylas i slammet; b) eventuellt tillsätter en effektiv mängd av en 15 antioxidant i slammet; och c) läter slammet reagera under en lämplig tid och vid en lämplig temperatur för att överföra stärkelsen i flytande form; varvid det muterade alfa-amylaset väljs ur gruppen 20 bestäende av: a) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas frän Bacillus licheniformis genom punktspeci- 25 fik substitution av en aminosyra i position M+15, och b) ett alfa-amylas som är expressionsprodukten av o en muterad DNA-sekvens som kodar ett alfa-amylas, varvid CM ^ den muterade DNA-sekvensen härstammar frän ett prekursor- ° alfa-amylas, som är ett Bacillus-alfa-amylas, genom punkt- oo 30 specifik substitution av en metioningrupp vars position x £ antingen i primär- eller tertiärstrukturen motsvarar M+15 LO i Bacillus licheniformis alfa-amylas, o § varvid nämnda alfa-amylas uppvisar en förändrad 00 o pH- och/eller temperaturverknmgsprofil i jämförelse med ^ 35 alfa-amylas hos Bacillus av vildtyp; varvid den ersättande aminosyran är Leu, Asn, Asp, Ser, Vai eller Ile.
23. En komposition som överför stärkelse i fly-tande form, vilken komposition innehäller muterat alfa-amylas, varvid nämnda muterade alfa-amylas valts ur grup-pen bestäende av: 5 a) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas frän Bacillus licheniformis genom punktspeci-fik substitution av en aminosyra i position M+15, ooh 10 b) ett alfa-amylas som är expressionsprodukten av en muterad DNA-sekvens som kodar ett alfa-amylas, varvid den muterade DNA-sekvensen härstammar frän ett prekursor-alfa-amylas, som är ett Bacillus-alfa-amylas, genom punkt-specifik substitution av en metioningrupp vars position 15 antingen i primär- eller tertiärstrukturen motsvarar M+15 i Bacillus licheniformis alfa-amylas, varvid nämnda alfa-amylas uppvisar en förändrad pH- och/eller temperaturverkningsprofil i jämförelse med alfa-amylas hos Bacillus av vildtyp; varvid den ersättande 20 aminosyran är Leu, Asn, Asp, Ser, Vai eller Ile.
24. En komposition enligt patentkrav 23, där nämnda muterade alfa-amylas är M15L. C\J δ CM CO o CO X cc CL LO O o o 00 o o CM
FI20080005A 1993-02-11 2008-01-07 Oxidationsbeständig alfaamylas FI122574B (sv)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US1639593A 1993-02-11 1993-02-11
US1639593 1993-02-11
US9401553 1994-02-10
PCT/US1994/001553 WO1994018314A1 (en) 1993-02-11 1994-02-10 Oxidatively stable alpha-amylase

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FI20080005A FI20080005A (sv) 2008-01-07
FI122574B true FI122574B (sv) 2012-03-30

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ID=21776903

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FI953797A FI119694B (sv) 1993-02-11 1995-08-10 Oxidationsbeständig alfaamylas
FI20080005A FI122574B (sv) 1993-02-11 2008-01-07 Oxidationsbeständig alfaamylas

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Application Number Title Priority Date Filing Date
FI953797A FI119694B (sv) 1993-02-11 1995-08-10 Oxidationsbeständig alfaamylas

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US (3) US6297037B1 (sv)
EP (2) EP0867504B2 (sv)
JP (1) JPH08506491A (sv)
KR (1) KR100322793B1 (sv)
CN (1) CN1104499C (sv)
AT (2) ATE175235T1 (sv)
AU (1) AU682863C (sv)
BR (1) BR9405720A (sv)
CA (1) CA2155831C (sv)
CZ (1) CZ293163B6 (sv)
DE (2) DE69415659T3 (sv)
DK (2) DK0689589T4 (sv)
ES (2) ES2198617T5 (sv)
FI (2) FI119694B (sv)
HU (1) HU219675B (sv)
NO (1) NO307187B1 (sv)
NZ (1) NZ262623A (sv)
PL (1) PL310326A1 (sv)
PT (1) PT867504E (sv)
WO (1) WO1994018314A1 (sv)

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