DK2554666T3 - Fremgangsmåde til opnåelse af ethanol uden glucoamylase ved anvendelse af pseudomonas saccharophila g4-amylase og varianter deraf - Google Patents

Fremgangsmåde til opnåelse af ethanol uden glucoamylase ved anvendelse af pseudomonas saccharophila g4-amylase og varianter deraf Download PDF

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DK2554666T3
DK2554666T3 DK12190129.2T DK12190129T DK2554666T3 DK 2554666 T3 DK2554666 T3 DK 2554666T3 DK 12190129 T DK12190129 T DK 12190129T DK 2554666 T3 DK2554666 T3 DK 2554666T3
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Andrew Shaw
Regina Chin
Karsten Matthias Kragh
Anja Hemmingsen Kellet-Smith
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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
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • A23L29/35—Degradation products of starch, e.g. hydrolysates, dextrins; Enzymatically modified starches
    • 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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Claims (9)

  1. FREMGANGSMÅDE TIL OPNÅELSE AF ETHANOL UDEN GLUCOAMYLASE VED ANVENDELSE AF PSEUDOMONAS SACCHAROPHILA G4-AMYLASE OG VARIANTER DERAF PATENTKRAV
    1. Fremgangsmåde til bearbejdning af stivelse omfattende flydendegørelse af en stivelse og/eller forsukring af et stivelsesforflydning for at danne en saccharidsirup ved tilsætning af en Pseudomonas saccharophila-amyl&se- (PS4) variant, hvor varianten omfatter en sekvens med mindst 90 % sekvensidentitet med aminosyresekvensen ifølge SEQ ID NO: 1, resterne 1 til 429 ifølge SEQ ID NO: 1, eller SEQ ID NO: 2, hvor varianten har et reduceret forhold mellem exo- og endo-amylaseaktivitet sammenlignet med aminosyresekvensen ifølge SEQ ID NO: 1, resterne 1 til 429 ifølge SEQ ID NO: 1 eller SEQ ID NO: 2, og hvor PS4-varianten omfatter aminosyresubstitutionen Q169R, Q169K eller Q169V ved en position svarende til positionen 169 ifølge SEQ ID NO: 1, og eventuelt; (i) én eller flere aminosyresubstitutioner ved følgende positioner: 7, 8, 32, 38, 49, 62, 63, 64, 67, 72, 73, 74, 75, 76, 104, 106, 107, 110, 112, 116, 119, 122, 123, 124, 125, 126, 128, 130, 137, 138, 140, 142, 143, 144, 148, 149, 150, 151, 154, 156, 163, 164, 168, 182, 183, 192, 195, 196, 200, 202, 208, 213, 220, 222, 225, 226, 227, 232, 233, 234, 236, 237, 239, 253, 255, 257, 260, 264, 267, 269, 271, 276, 282, 285, 295, 297, 300, 302, 305, 308, 312, 323, 324, 325, 341, 358, 367, 379, 390 ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; (ii) én eller flere af følgende aminosyresubstitutioner: A3T, G9A, H13R, I46F, D68E, G69A/E/H/I/K/M/R/T, G70A/E/L/P/Q/S/V, K71M, G100A/S, G121I/P/R, A131T, G134C, A141S, N145S, Y146D/E, G153A/D, G158C/F/I/L/P/Q/V, S161G/II/K/P/R/T/V, G166N, I170E/K/L/M/N, L178N/Q/W, A179E/N/P/R/S, A179S, G184Q, G188A, A199P, G223C/F/II/M/N/Q/W/Y, S229N, W238E/G/K/P/Q/R, G303L, H307D/E/F/G/K/M/P/Q/R/S/W/Y, A309E/I/M/T/V, S334A/H/K/L/M/Q/R/T og/eller H335M ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; og/eller (iii) én eller flere aminosyresubstitutioner ved positionerne 420, 422 og/eller 424 ifølge SEQ ID NO: 1.
  2. 2. Fremgangsmåde ifølge krav 1, hvor PS4-varianten omfatter en sekvens med mindst 95 % sekvensidentitet med aminosyresekvensen ifølge SEQ ID NO: 1, resterne 1 til 429 ifølge SEQ ID NO: 1 eller SEQ ID NO: 2.
  3. 3. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 2, hvor PS4-varianten omfatter én eller flere aminosyresubstitutioner ved følgende positioner: A3T, P7S, A8N, G9A, H13R, P32S, I38M, I46F, D49V, D62N, F63A/D/E/L/V, S64N/T, T67G/H/K/N/Q/R/V, D68E, G69 A/E/H/I/K/M/R/T, G70A/E/L/P/Q/S/V, K71M, S72E/K/N/T, G73D/E/L/M/N/S/T, G74S, G75C/E/F/R/S/W/Y, E76V, G100A/S, G104N/R, G106K, V107M, L110F, D112E, N116D, N119E/G/S/Y, G121I/P/R, Y122A/E/Q/W, P123S, D124S, K125A/D/E/G/P/Q/W, E126D/N, N128E, P130S, A131T, G134C, R137C, N138D/E/S, C140A/R, A141S, D142E/G/N, P143T, G144E, N145S, Y146DÆ, N148K/S, D149H/L/V, C150A, D151A/V/W, G153A/D, D154E/G/Y, F156Y, GI58C/F/I/L/P/Q/V, S161G/H/K/P/R/T/V, L163M,N164R, G166N, P168L, 1170E/K/L/M/N, L178N/Q/W, A179E/N/P/R/S, R182D/G/H/M/S, S183G, G184Q, G188A, F192M/Y, V195D, R196A/G/K/P/Q/S/T/V/Y, A199P, P200A/G, R202K, S208T, S213N, L220A/T, K222M/Y, G223C/F/II/M/N/Q/W/Y, S225E/G/V, E226C/D/G/W, Y227C/D/G/K/T, S229N, W232F/G/H/I/K/L/N/P/Q/R/S/T/Y, R233H, N234R, A236E, S237D/G, W238E/G/K/P/Q/R, Q239L, V253G, D255V, A257V, E260K/R, N264D, V267I, D269N/S/V, K271A/L/Q, G276R, W282S, V285A, T295C, Y297H, G300E, N302K, G303L, Q305E/L/T, H307D/E/F/G/K/M/P/Q/R/S/W/Y, W308A/C/G/K/N/O/R/S/T, A309E/I/M/T/V, D312E, W323M, T324A/L/M, S325G, S334A/H/K/L/M/Q/R/T, H335M, Y341C/E, R358A/E/G/L/N/0/T/V, S367Q/R, S379G og/eller D390E ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; og/eller én eller flere substitutioner af S420G, D422N/P/Q og/eller G424D/S ifølge SEQ ID NO: 1.
  4. 4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 2, hvor PS4-varianten omfatter én eller flere aminosyresubstitutioner ved følgende positioner: 7, 32, 49, 62, 63, 64, 72, 73, 74, 75, 76, 107, 110, 112, 116, 119, 122, 123, 125, 128, 130, 137, 138, 140, 142, 143, 144, 148, 149, 150, 151, 154, 156, 163, 164, 168, 182, 183, 192, 195, 196, 202, 220, 222, 226, 227, 232, 233, 234, 236, 237, 239, 253, 255, 257, 260, 264, 269, 271, 276, 282, 285, 297, 300, 302, 305, 308, 312, 323, 324, 325, 341, 358, 367 og/eller 379 ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; én eller flere af følgende aminosyresubstitutioner: A3T, H13R, I38M, I46F, T67G/H/K/N/Q/R/V, G69A/E/H/I/K/M/R/T, G70E/L/P/Q/V, K71M, G100A/S, G104R, G106K, G121I/P/R, D124S, E126D/N, A131T, G134C, A141S, N145S, Y146D/E, G153A/D, G158C/F/I/L/P/Q/V, S161G/H/K/P/R/T/V, G166N, 1170E/K/L/M/N, L178N/Q/W, A179E/N/P/R/S, G188A, A199P, P200A, G223C/F/H/M/N/Q/W/Y, S225E/G/V, W23 8E/G/K/P/Q/R, T295C, G303L, H307D/G/M/P/S, A309E/I/M/T/V, S334A/H/K/L/M/Q/R/T, H335M og/eller D390E ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; én eller flere aminosyresubstitutioner S420G og/eller D422/N/P/Q ifølge SEQ ID NO: 1; og/eller en aminosyresubstitution ved position 424 ifølge SEQ ID NO: 1.
  5. 5. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 2, hvor PS4-varianten omfatter én eller flere aminosyresubstitutioner ved følgende positioner: 49, 62, 63, 64, 72, 73, 74, 75, 76, 107, 112, 116, 119, 122, 123, 125, 128, 130, 137, 140, 143, 144, 148, 149, 150, 151, 154, 156, 163, 164, 168, 182, 183, 192, 195, 196, 202, 257, 282, 285, 297, 300, 305, 308, 312, 323 og/eller 325 ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; én eller flere af følgende aminosyresubstitutioner: A3T, P7S, H13R, I38M, I46F, T67G/H/K/N/Q/R/V, G69A/E/H/I/K/M/R/T, G70E/L/P/Q/V, K71M, G100A/S, G104R, G106K, L110F, G121I/P/R, D124S, E126D/N, A131T, G134C, N138D/E, D142/E/G/N, N145S, Y146D/E, G153A/D, G158C/F/I/L/P/Q/V, S161G/H/K/P/R/T/V, G166N, I170E/K/L/M, L178N/Q/W, A179E/N/P/R/S, G188A, A199P, P200A, L220T, K222M/Y, G223C/F/H/M/N/Q/W/Y, S225E/V, E226C/D/G/W, Y227C/D/G/K/T, W232F/G/H/I/K/N/P/Q/R/S/T/Y, R233H, N234R, A236E, S237D/G, W238E/G/K/P/Q/R, Q239L, V253G, D255V, E260K/R, N264D, D269N/S/V, K271A/L/Q, G276R, T295C, N302K, G303L, H307D/G/M/P/S, A309E/I/M/T/V, T324L/M, S334A/H/K/L/M/Q/R/T, H335M, Y341C/E, R358A/E/G/L/N/Q/T/V, S367Q/R, S379G og/eller D390E ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6; og én eller flere af følgende aminosyresubstitutioner: S420G, D422/N/P/Q og/eller G424S ifølge SEQ ID NO: 1.
  6. 6. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 5, hvor PS4-varianten omfatter én eller flere aminosyresubstitutioner ved følgende positioner: N33, D34, G70, G121, G134, A141, Y146, 1157, S161, L178, A179, G223, S229, H307, A309 og/eller S334 iMge SEQ ID NO: 1 eller 2.
  7. 7. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 6, hvor PS4-varianten har op til 25 aminosyredeletioner, -additioner, -indsætninger eller substitutioner sammenlignet med aminosyresekvensen ifølge SEQ ID NO: 1, 2, 3, 4, 5 eller 6.
  8. 8. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 7, hvor: (i) fremgangsmåden endvidere omfatter tilsætning af et frakoblende enzym, en isoamylase, en pullulanase, en protease, en cellulase, en hemicellulase, en lipase, en cutinase eller en hvilken som helst kombination af enzymerne til stivelsesforflydning; og/eller (ii) stivelsen er fra majs, majskolber, hvede, byg, rug, hirse, sago, kassava, tapioka, durra, ris, ærter, bønner, banan eller kartofler.
  9. 9. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 8, hvilken fremgangsmåde endvidere omfatter fermentering af saccharidsiruppen for at fremstille ethanol, hvilken fremgangsmåde eventuelt endvidere omfatter genvinding af ethanol, eventuelt endvidere omfatter destillering af stivelsen for at opnå ethanol, hvor fermentering og destillering udføres samtidigt, separat eller sekventielt.
DK12190129.2T 2008-01-02 2008-12-30 Fremgangsmåde til opnåelse af ethanol uden glucoamylase ved anvendelse af pseudomonas saccharophila g4-amylase og varianter deraf DK2554666T3 (da)

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US624008P 2008-01-02 2008-01-02
EP12187500A EP2546338A1 (en) 2008-01-02 2008-12-30 A process of obtaining ethanol without glucoamylase using pseudomonas saccharophila g4-amylase and variants thereof

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US20090311764A1 (en) 2009-12-17
WO2009088465A3 (en) 2009-09-03
US20110033575A1 (en) 2011-02-10
CN102943098A (zh) 2013-02-27
US8318451B2 (en) 2012-11-27
EP2546338A1 (en) 2013-01-16
EP2238244A1 (en) 2010-10-13
WO2009088465A2 (en) 2009-07-16
CN101918545A (zh) 2010-12-15
EP2554666A1 (en) 2013-02-06
WO2009083592A1 (en) 2009-07-09
EP2554666B1 (en) 2015-08-05
CN102978258A (zh) 2013-03-20

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