DK2337837T4 - Alpha-amylase blandinger og fremgangsmåder til anvendelse deraf - Google Patents

Alpha-amylase blandinger og fremgangsmåder til anvendelse deraf Download PDF

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DK2337837T4
DK2337837T4 DK09792446.8T DK09792446T DK2337837T4 DK 2337837 T4 DK2337837 T4 DK 2337837T4 DK 09792446 T DK09792446 T DK 09792446T DK 2337837 T4 DK2337837 T4 DK 2337837T4
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amylase
alpha
starch
amino acid
amys
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DK2337837T3 (da
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Bradley A Paulson
Vivek Sharma
Jayarama Shetty
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Danisco Us Inc
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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
    • C11D3/38681—Chemically modified or immobilised enzymes
    • 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
    • C12P19/02—Monosaccharides
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
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  • Genetics & Genomics (AREA)
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  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
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  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Detergent Compositions (AREA)

Claims (17)

  1. ALPHA-AMYLASE BLANDINGER OG FREMGANGSMÅDER TIL ANVENDELSE DERAF PATENTKRAV
    1. Fremgangsmåde til flydendegørelse af stivelse, hvilken fremgangsmåde omfatter: tilføjelse af en amylase-blanding til en opløsning omfattende stivelse og flydendegørelse af opløsningen, der omfatter stivelse, for at danne en opslæmning, hvor amylase-blandingen omfatter: (i) en B. stearothermophilus alpha-amylase (AmyS), der omfatter en amino syre sekvens med mindst 85 % sekvensidentitet med AmyS ifølge SEQ ID NO: 1, hvor AmyS har alpha-amylase-aktivitet, og hvor aminosyren ved position S242 er substitueret, ved anvendelse af aminosyretalsystemet ifølge SEQ ID NO: 2, hvor substitutionen er S242Q eller S242A; og (ii) en IS. licheniformis alpha-amylase, der omfatter en aminosyresekvens: ANLNGTLMQY FEWYTPNDGQ HWKRLQN DS A YLAEHGITAV WIPPAYKGTS QADVGYGAYD LYDLGEFHQK GTVRTKYGTK GELQSAIKSL HSRDINVYGD VV1NHKGGAD ATEDVTAVEV DPADRNRVIS GEYLIKAWI H FHFPGRGSTY SDFKWHWYHF DGTDWDESRK LNRIYKFQGK AWDWEVSSEN GNYDYLMYAD IDYDHPDVVA E1KRWGTWYA NELQLDGFRL DAVKHIKFSF LRDWVNHVRE KTGKEMFTVA EYWQNDLGAL ENYLNKTNFN HSVFDVPLHY QFHAASTQGG GYDMRKLLNG TVVSKHPLKS VTFVDNHDTQ PGQSLESTVQ TWFKPLAYAF ILTRESGYPQ VFYGDMYGTK GDSQREIPAL KHKIEPILKA RKQYAYGAQH DYFDHHDIVG WTREGDSSVA NSGLAALITD GPGGAKRMYV GRQNAGETWH DITGNRSEPV VINSEGWGEF HVNGGSVSIY VQR.
  2. 2. Fremgangsmåde ifølge krav 1, hvor stivelsen opnås fra plantemateriale, majs, milokom, mg, byg, hvede, durra, havre, ris, klid, kassava, hirse, kartoffel, sød kartoffel eller tapioka, og eventuelt hvor stivelsen er granulær stivelse fra enten en tør eller våd formalingsmetode.
  3. 3. Fremgangsmåde ifølge krav 1 eller krav 2, hvilken fremgangsmåde endvidere omfatter: (i) et primært og/'eller sekundært flydendegørelsestrin, herunder tilføjelse af yderligere substrat til opslæmningen i det primære og/'eller sekundære flydendegørelsestrin; og/eller (ii) tilføjelse af et fytinsyrehydrolyserende enzym; og/eller (iii) saccharifrcering af den flydende stivelse for at opnå gæringsdygtigt sukker, og eventuelt gæring af det gæringsdygtige sukker under egnede gæringsforhold for at opnå alkoholiske slutprodukter.
  4. 4. Fremgangsmåde ifølge krav 3 (iii), hvor de alkoholiske slutprodukter er ethanol eller butanol.
  5. 5. Fremgangsmåde ifølge krav 1, hvor: (i) flydendegørelsens pH ligger i intervallet fra pH 4,5 til 5,4, og syreneutraliserende kemikalier ikke tilføjes i trinnet med flydendegørelsesprocessen; eller (ii) flydendegørelsens pH ligger i intervallet fra pH 4,8 til 5.8, og syreneutraliserende kemikalier ikke tilføjes i trinnet med flydendegørelsesprocessen.
  6. 6. Fremgangsmåde til opnåelse af et gæringsdygtigt substrat, hvilken fremgangsmåde omfatter: etablering af kontakt mellem en opslæmning af malet kom, der indeholder granulær stivelse, med både et fytinsyrehydrolyserende enzym og en blanding af alpha-amylaser ved en temperatur på 0-30 °C lavere end stivelsens gelatineringstemperatur, hævelse af temperaturen til over gelatineringstemperaturen, hydrolysering af den gelatinerede stivelse og opnåelse af et gæringsdygtigt substrat, hvor blandingen af alpha-amylaser omfatter: (i) en B. stearothermophilus alpha-amylase (AmyS), der omfatter en aminosyresekvens med mindst 85 % sekvensidentitet med AmyS ifølge SEQ ID NO: 1, hvor AmyS har alpha-amylase-aktivitet, og hvor aminosyren ved position S242 er substitueret, ved anvendelse af aminosyretalsystemet ifølge SEQ ID NO: 2, hvor substitutionen er S242Q eller S242A; og (ii) en B. licheniformis alpha-amylase, der omfatter en aminosyresekvens: ANLNGTLMQY FEWYTPNDGQ HWKRLQNDSA YLAEHGITAV WIPPAYKGTS QADVGYGAYD LYDLGEFHQK GTVRTKYGTK GELQSAIKSL HSRDINVYGD VVINHKGGAD ATEDVTAVEV DPADRNRVIS GEYLIKAWTH FHFPGRGSTY SDFKWHWYHF DGTDWDESRK LNRIYKFQGK AWDWEVSSEN GNYDYLMYAD 1DYDHPDVVA EIKRWGTWYA NELQLDGFRL DAVKHIKFSF LRDWVNHVRE KTGKEMFTVA EYWQNDLGAL ENYLNKTNFN HSVFDVPLHY QFHAASTQGG GYDMRKLLNG TWSKHPLKS VTFVDNHDTQ PGQSLESTVQ TWFKPLAYAF ILTRESGYPQ VFYGDMYGTK. GDSQREIPAL K.HKIEPILKA RKQYAYGAQH DYFDHHDIVG WTREGDSSVA NSGLAAL1TD GPGGAKRMYV GRQNAGETWH DITGNRSEPV VINSEGWGEF HVNGGSVSIY VQR.
  7. 7. Fremgangsmåde til fremstilling af et gæringsdygtigt sukker, hvilken fremgangsmåde omfatter: (a) blanding af malet stivelsesholdigt materiale med vand og bærme, hvor bærmen ligger i intervallet fra 10 til 70 % volumen/volumen, og opnåelse af en opslæmning, der omfatter stivelse og har et tørstofindhold (ds) på 20 til 50 % vægt/vægt, (b) behandling af opslæmningen med en fytase før eller samtidig med flydendegørelse af stivelsen, (c) flydendegørelse af stivelsen, (d) tilføjelse en alpha-amylase-blanding til stivelsen enten under trin (b) og/eller samtidig med trinnet med flydendegørelse, og (e) saccharificering af den flydende stivelse for at opnå gæringsdygtige sukkere, hvor pH’et ikke er justeret under nogen som helst af trinnene (a), (b), (c), (d) eller (e), hvor alpha-amylase-blandingen omfatter: (i) en B. stearothermophilus alpha-amylase (AmyS), der omfatter en aminosyresekvens med mindst 85 % sekvensidentitet med AmyS ifølge SEQ ID NO: 1, hvor AmyS har alpha-amylase-aktivitet, og hvor aminosyren ved position S242 er substitueret, ved anvendelse af aminosyretalsystemet ifølge SEQ ID NO: 2, hvor substitutionen er S242Q eller S242A; og (ii) en B. licheniformis alpha-amylase, der omfatter en aminosyresekvens: ANLNGTLMQY FEWYTPNDGQ HWKRLQNDSA YLAEHGITAV WIPPAYKGTS QADVGYGAYD LYDLGEFHQK GTVRTKYGTK GELQSAIKSL HSRDINVYGD VVINHKGGAD ATEDVTAVEV DPADRNRVIS GEYL1XAWTH FHFPGRGSTY SDFKWHWYHF DGTDWDESRK LNR1YKFQGK AWDWEVSSEN GNYDYLMYAD IDYDFIPDWA E1KRWGTWYA NELQLDGFRL DAVKHIKFSF LRDWVNHVRE KTGKEMFTVA EYWQNDLGAL ENYLNKTNFN HSVFDVPLHY QFHAASTQGG GYDMRKLLNG TVVSKIIPLKS VTFVDNHDTQ PGQSLESTVQ TWFKPLAYAF ILTRESGYPQ VFYGDMYGTK GDSQREIPAL KHKIEPILKA RKQYAYGAQH DYFDHHDIVG WTREGDSSVA NSGLAALITD GPGGAKRMYV GRQNAGETWH D1TGNRSEPV VINSEGWGEF HVNGGSVSTY VQR.
  8. 8. Fremgangsmåde ifølge krav 7, hvor: (i) det gæringsdygtige sukker er fjernet og oprenset eller isomeriseret; og/eller (ii) fytasen tilsættes før trinnet med flydendegørelse; og/eller (iii) alpha-amylase-blandingen tilsættes sammen med fytasen; og/eller (iv) en anden dosis af alpha-amylase-blandingen tilsættes under trinnet med flydendegørelse eller tilsættes under trinnet med saccharificering.
  9. 9. Fremgangsmåde til fremstilling af glucose, hvilken fremgangsmåde omfatter; etablering af kontakt mellem et stivelsessubstrat med en amylase-blanding, flydendegørelse af stivelsessubstratet, og inkubation af substratet ved høj temperatur for at fremstille et produkt, der omfatter glucose, hvor amylase-blandingen omfatter: (i) en B. stearothermophilus alpha-amylase (AmyS), der omfatter en aminosyresekvens med mindst 85 % sekvensidentitet med AmyS ifølge SEQ ID NO: 1, hvor AmyS har alpha-amylase-aktivitet, og hvor aminosyren ved position S242 er substitueret, ved anvendelse af aminosyretalsystemet ifølge SEQ ID NO: 2, hvor substitutionen er S242Q eller S242A; og (ii) en B. licheniformis alpha-amylase, der omfatter en aminosyresekvens: ANLNGTLMQY FEWYTPNDGQ HWKRLQNDSA YLAEHGITAV WIPPAYKGTS QADVGYGAYD LYDLGEFHQK GTVRTKYGTK GELQSAIKSL HSRDINVYGD VVINHKGGAD ATEDVTAVEV DPADRNRVIS GEYLIKAWTH FHFPGRGSTY SDFKWHWYHF DGTDWDESRK LNR1YKFQGK AWDWEVSSEN GNYDYLMYAD 1DYDHPDVVA ElKRWGTWYA NELQLDGFRL DAVKHIKFSF LRDWVNHVRE KTGKEMFTVA EYWQNDLGAL ENYLNKTNFN HSVFDVPLHY QFHAASTQGG GYDMRKLLNG TVVSKHPLKS VTFVDNHDTQ PGQSLESTVQ TWFKPLAYAF ILTRESGYPQ VFYGDMYGTK GDSQREIPAL KHKIEPILKA RKQYAYGAQH DYFDHHDIVG WTREGDSSVA NSGLAALITD GPGGAKRMYV GRQNAGETWH DITGNRSEPV VINSEGWGEF HVNGGSVSIY VQR.
  10. 10. Fremgangsmåde ifølge krav 9, hvor: (i) inkubationstrinnet er et sekundært trin med flydendegørelse; og/eller (ii) inkubationstrinnet udføres ved en temperatur på ca. 90-100 °C; og/eller (iii) inkubationen udføres i tilstrækkelig tid, til at produktet omfatter ca. 2-14 DE glucose, og eventuelt hvor inkubationen udføres i tilstrækkelig tid, til at produktet omfatter ca. 10 DE glucose.
  11. 11. Fremgangsmåde ifølge krav 9, hvilken fremgangsmåde endvidere omfatter saccharificcring af produktet for at fremstille en glucose-rig opløsning.
  12. 12. Fremgangsmåde ifølge krav 11, hvor den glueose-rige opløsning omfatter 80-99 % glucose, fortrinsvis hvor den glueose-rige opløsning omfatter ca. 93-96 % glucose.
  13. 13. Fremgangsmåde ifølge krav 11, hvor saccharificeringen omfatter etablering af kontakt mellem glucoseproduktet og glueoamylase eller en enzymblanding, der omfatter en glueoamylase og en pullulanase.
  14. 14. Alpha-amylase-blanding, der omfatter: (i) en B. stearothermophilus alpha-amylase (AmyS), der omfatter en aminosyresekvens med mindst 85 % sekvensidentitet med AmyS ifølge SEQ ID NO: 1, hvor AmyS har alpha-amylase-aktivitet, og hvor aminosyren ved position S242 er substitueret, ved anvendelse af aminosyretalsystemet ifølge SEQ ID NO: 2, hvor substitutionen er S242Q eller S242A; og (ii) en B licheniformis alpha-amylase, der omfatter en aminosyresekvens: ANLNGTLMQY FEWYTPNDGQ HWKRLQNDSA YLAEHG1TAV WfPPAYKGTS QADVGYGAYD LYDLGEFHQK GTVRTKYGTK GELQSAIKSL HSRDINVYGD WINHKGGAD ATEDVTAVEV DPADRNRVIS GEYLIKAWTH FHFPGRGSTY SDFKWFIWYHF DGTDWDESRK LNR1YKFQGK AWDWEVSSEN GNYDYLMYAD IDYDHPDVVA EIKRWGTWYA NELQLDGFRL DAVKH1KFSF LRDWVNHVRE KTGKEM.FTVA EYWQNDLGAL ENYLNKTNFN HSVFDVPLHY QFHAASTQGG GYDMRKLLNG TVVSKHPLKS VTFVDNHDTQ PGQSLESTVQ TWFKPLAYAF ILTRESGYPQ VFYGDMYGTK GDSQREIPAL KHKIEP1LKA RKQYAYGAQH DYFDHHD1VG WTREGDSSVA NSGLAALITD GPGGAKRMYV GRQNAGETWH DITGNRSEPV V1NSEGWGEF HVNGGSVS1Y VQR.
  15. 15. Alpha-amylaseblanding ifølge krav 14, hvilken blanding endvidere omfatter en fytase.
  16. 16. Alpha-amylaseblanding ifølge krav 14 eller krav 15, hvor blandingen omfatter et vægtforhold på ca. 40 % af AmyS med S242-substitutionen og ca. 60 % B. licheniformis alpha-amylase.
  17. 17. Alpha-amylaseblanding ifølge krav 14 eller krav 15, hvor vægtforholdet mellem AmyS og S242-substitutionen og /1. licheniformis alpha-amylase er 10:90.
DK09792446.8T 2008-09-25 2009-09-11 Alpha-amylase blandinger og fremgangsmåder til anvendelse deraf DK2337837T4 (da)

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PCT/US2009/056613 WO2010036515A1 (en) 2008-09-25 2009-09-11 Alpha-amylase blends and methods for using said blends

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CA2738447A1 (en) 2010-04-01
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JP2012503484A (ja) 2012-02-09
EP2337837B1 (en) 2013-04-03
MX2011003178A (es) 2011-04-21
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