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DK1781790T3 - Alfa-amylasevarianter med ændrede egenskaber - Google Patents

Alfa-amylasevarianter med ændrede egenskaber Download PDF

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DK1781790T3
DK1781790T3 DK05755531.0T DK05755531T DK1781790T3 DK 1781790 T3 DK1781790 T3 DK 1781790T3 DK 05755531 T DK05755531 T DK 05755531T DK 1781790 T3 DK1781790 T3 DK 1781790T3
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Carsten Andersen
Henrik Østdal
Peter Skagerlind
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/24Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405Glucanases
    • C12N9/2408Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411Amylases
    • C12N9/2414Alpha-amylase (3.2.1.1.)
    • C12N9/2417Alpha-amylase (3.2.1.1.) from microbiological source
    • CCHEMISTRY; METALLURGY
    • C11ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11DDETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16Organic compounds
    • C11D3/38Products with no well-defined composition, e.g. natural products
    • C11D3/386Preparations containing enzymes, e.g. protease or amylase
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/14Preparation 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
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01001Alpha-amylase (3.2.1.1)
    • DTEXTILES; PAPER
    • D06TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06MTREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M16/00Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
    • D06M16/003Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic with enzymes or microorganisms
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2299/00Coordinates from 3D structures of peptides, e.g. proteins or enzymes

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Textile Engineering (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Claims (31)

1. Variant af en Termamyl-lignende stam-alfa-amylase, hvor varianten har alfa-amylaseaktivitet, og hvor varianten omfatter substitutionerne Y295F og M202L,I,T,V, hvor hver position svarer til en position i aminosyresekvensen ifølge SEQ ID NO: 12, hvor varianten har øget stabilitet over for oxidation i forhold til stam-alfa-amylasen, og hvor alfa-amylasevarianten udviser mindst 90% identitet med aminosyresekvensen ifølge SEQ ID NO: 12.
2. Variant ifølge krav 1, som endvidere omfatter en ændring i en eller flere positioner valgt fra gruppen bestående af: 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 203, 214, 231,256, 257, 258, 269, 270, 272, 283, 296, 298, 299, 303, 304, 305, 311,314, 315, 318, 319, 339, 345, 361, 378, 383, 419, 421,437, 441,444, 445, 446, 447, 450, 461,471,482 og 484.
3. Variant ifølge krav 2, hvor mutationerne er: R26S, D30N, N33D, R82H, K37T, N106D, K118Q, N128Y, G133E,A, G149A,N, N150H,Q, Y160F, Y178F, G182T, G186A, T193S,N,D,E,Q, Y203L, V214I.T, D231N, G256K, T257I, G258D, K269S,Q, N270F,Y,D, L272I,V,A, N283D, N296K,Q,E, Y298F,H, N299F,Y,Q,T, S303Q,K, Y304F,R,K, G305D, Q311 N,Q,K,R,T,S,Y,F, N314D,S,T,Q, G315N,S,T, V318L, Q319E,K,S,T, A339S,T, E345N,R, Q361E, G378K, K383R, T419N, H421Y, N437H, F441L, R444E,Y, N445Q, K446R, A447Y, V450T, T461P, N471E, W482Y, N484Q.
4. Variant ifølge krav 1-3, hvor varianten har en yderligere mutation i en eller flere methioninrester.
5. Variant ifølge krav 4, hvor methioninresterne er: M9, M10, M116, M208, M261, M309, M323, M382, M410, M430 og M440.
6. Variant ifølge krav 5, hvor mutationerne er: M9L, I, M10L, M105L,I,F, M116N,D,L,I,F,W,R,K, M208F,Y,L,I, M261 L,l, M309L,I, M323L,I,S,T,A,Q,E,N,D, M382L,I,Y,F,K, M410L,I,V, M430L,I og M440L,I,F,Y.
7. Variant ifølge krav 1-6, hvor varianten haren eller flere af følgende mutationer: M9L+M202I+Y295F+A339S, M9L+M202I+Y295F, M9L+M202I+Y295F+A339S, M9L+M202I+Y295F+A339S+E345R, M9L+G149A+M202I+Y295F+A339S+E345R, M9L+M202L+Y295F+A339S, M9L+M202L+Y295F, M9L+M202L+Y295F+A339S, M9L+M202L+Y295F+A339S+E345R, M9L+G149A+M202L+Y295F+A339S+E345R, M9L+M202T+Y295F+A339S, M9L+M202T+Y295F, M9L+M202T+Y295F+A339S, M9L+M202T+Y295F+A339S+E345R, M9L+G149A+M202T+Y295F+A339S+E345R, M9L+G149A+M202I+V214T+Y295F+N299Y+M323T+A339S+E345R, M9L+G149A+M202L+V214I+Y295F+M323T+A339S+E345R+M382Y, M9L+G149A+G182T+G186A+M202I+V214I+Y295F+N299Y+M323T+ A339S, M9L+G149A+G182T+G186A+M202L+T257I+Y295F+N299Y+M323T+A339S+E345R, M9L+G149A+M202L+V214T+Y295F+N299Y+M323T+A339S+E345R, M9L+G149A+M202I+V214I+Y295F+M323T+A339S+E345R+M382Y, M9L+G149A+G182T+G186A+M202L+V214I+Y295F+N299Y+M323T+A339S, M9L+G149A+G182T+G186A+M202I+T257I+Y295F+N299Y+M323T+A339S+E345R, M9L+G149A+M202I+V214Τ+Y295F+N299Y+M323T+A339S+E345R+N471Ε, M9L+G149A+M202L+V214I+Y295F+M323T+A339S+E345R+M382Y+N471Ε, M9L+G149A+G182T+G186A+M202I+V214I+Y295F+N299Y+M323T+A339S+N471 Ε, M9L+G149A+G182Τ+G186A+M202L+T257I+Y295F+N299Y+M323T+A339S+E345R+N471 Ε, M202L+Y295F, M202I+Y295F, M202V+Y295F, M202T+Y295F.
8. Variant ifølge krav 1-7, hvor varianten endvidere omfatter en deletion af D183+G184.
9. Variant ifølge krav 1-8, hvor varianten endvidere omfatter en mutation i R118, navnlig R118K.
10. Variant ifølge krav 1-9, hvor varianten endvidere omfatter en mutation i N195, navnlig N195F.
11. Variant ifølge krav 1-10, hvor varianten endvidere omfatter en mutation i R320, navnlig R320K.
12. Variant ifølge krav 1-11, hvor varianten endvidere omfatter en mutation i R458, navnlig R458K.
13. Variant ifølge krav 1-12, hvor varianten endvidere omfatter deletionen af D183+G184 og mutationen R118K+N195F+R320K+R458K i kombination med en eller flere af følgende mutationer: K118Q, K37T, H421Y, V450T, K383R, N445Q, Y178F, V318L, W482Y, N283D+Q361E, M105F+M208F K446R+N484Q, R444Y, N106D, Y203L, G133E+Q361E, M323E, V214T.
14. Variant ifølge krav 13, hvor varianten endvidere omfatter en eller flere af følgende mutationer: T461P, Y298H, G133E+R444E, Y298F, V214T+M323E+M382Y+K383R+N471E Y178F+G258D+T419N+N437H G149N+N150Q+M382Y+K383R Y160F+V214T+M382Y N128Y+G149A+V214T+D231N+M382Y+F441L R82H+N128Y+G149A+V214T+M382Y N150H+V214T V214T+E345N V214T+G305D+ M382Y+R444E V214T+M382Y+A447Y V214T+G378K V214T+A256K R26S+D30N+N33D+V214T+M382Y.
15. DNA-konstruktion, som omfatter en DNA-sekvens, der koder for en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14.
16. Rekombinant ekspressionsvektor, som bæreren DNA-konstruktion ifølge krav 15.
17. Celle, som er transformeret med en DNA-konstruktion ifølge krav 15 eller en vektor ifølge krav 16.
18. Celle ifølge krav 17, som er en mikroorganisme, fortrinsvis en bakterie eller en svamp, navnlig en grampositiv bakterie, såsom Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, Bacillus lautus eller Bacillus thuringiensis.
19. Sammensætning, som omfatter en alfa-amylasevariant ifølge krav 1-14.
20. Detergentadditiv, som omfatter en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14, og som eventuelt foreligger i form af et ikke-støvende granulat, stabiliseret væske eller beskyttet enzym.
21. Detergentadditiv ifølge krav 20, som indeholder 0,02-200 mg enzymprotein/g af additivet.
22. Detergentadditiv ifølge krav 20 eller 21, som endvidere omfatter et andet enzym, såsom en protease, en lipase, en peroxidase, mannanase, maltogen amylase, CGTase, amylase eller et andet amylolytisk enzym, såsom glucoamylase, og/eller en cellulase.
23. Detergentsammensætning, som omfatter en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14. 24. 15. Detergentsammensætning ifølge krav 23, som endvidere omfatter et andet enzym, såsom en protease, en lipase, en peroxidase, mannanase, maltogen amylase, CGTase, et andet amylolytisk enzym og/eller en cellulase.
25. Detergentsammensætning til manuel eller automatisk opvask, hvilken detergentsammensætning omfatter en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14.
26. Detergentsammensætning til opvask ifølge krav 25, hvilken detergentsammensætning endvidere omfatter et andet enzym, såsom en protease, en lipase, en peroxidase, mannanase, maltogen amylase, CGTase, amylase eller et andet amylolytisk enzym, såsom glucoamylase, og/eller en cellulase.
27. Sammensætning til manuel eller automatisk tøjvask, hvilken sammensætning omfatter en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14.
28. Sammensætning til tøjvask ifølge krav 27, hvilken sammensætning endvidere omfatter et andet enzym, såsom en protease, en lipase, en peroxidase, mannanase, maltogen amylase, CGTase, en amylase og/eller et andet amylolytisk enzym, såsom glucoamylase, og/eller en cellulase.
29. Anvendelse af en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14 eller en sammensætning ifølge krav 19-28 til vask og/eller opvask.
30. Anvendelse af en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14 eller en sammensætning ifølge krav 19 til afsletning af tekstil.
31. Anvendelse af en alfa-amylasevariant ifølge et hvilket som helst af kravene 1-14 eller en sammensætning ifølge krav 19 til forflydning af stivelse.
32. Fremgangsmåde til fremstilling af en variant ifølge et hvilket som helst af kravene 1-14, hvor en celle ifølge et hvilket som helst af kravene 17-18 dyrkes under betingelser, som er befordrende for fremstilling af varianten, og varianten efterfølgende indvindes fra kulturen.
DK05755531.0T 2004-07-05 2005-07-05 Alfa-amylasevarianter med ændrede egenskaber DK1781790T3 (da)

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PCT/DK2005/000469 WO2006002643A2 (en) 2004-07-05 2005-07-05 Alpha-amylase variants with altered properties

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EP (4) EP3620523A3 (da)
JP (2) JP4955546B2 (da)
CN (4) CN107151662B (da)
AU (1) AU2005259686B2 (da)
BR (1) BRPI0512776A (da)
CA (2) CA2854912A1 (da)
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