DK2662445T3 - Fremgangsmåde til at designe muteret enzym, fremgangsmåde til fremstilling af samme og muteret enzym - Google Patents

Fremgangsmåde til at designe muteret enzym, fremgangsmåde til fremstilling af samme og muteret enzym Download PDF

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Publication number
DK2662445T3
DK2662445T3 DK13179288.9T DK13179288T DK2662445T3 DK 2662445 T3 DK2662445 T3 DK 2662445T3 DK 13179288 T DK13179288 T DK 13179288T DK 2662445 T3 DK2662445 T3 DK 2662445T3
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atom
amino acid
enzyme
leu
tyra
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DK13179288.9T
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English (en)
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Hiroyuki Iwamoto
Bunzo Mikami
Shotaro Yamaguchi
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Amano Enzyme Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01041—Pullulanase (3.2.1.41)
    • 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
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • 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
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • C12N9/2457—Pullulanase (3.2.1.41)
    • 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
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • C12N9/246—Isoamylase (3.2.1.68)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01068—Isoamylase (3.2.1.68)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2299/00—Coordinates from 3D structures of peptides, e.g. proteins or enzymes

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (5)

1. Muteret enzym, hvor aktiviteten og/eller substratspecificiteten hvad angår dets Km-værdi og/eller Kcat-værdi for hydrolysering af en a-l,6-glycosidforbindelse i forhold til pullulan eller amylopektin i 50 mM acetatbuffer ved pH 5,6 ved 25 °C er forbedret sammenlignet med enzymet, der skal muteres, omfattende en aminosyresekvens i hvilken en eller to eller flere aminosyrer valgt fra gruppen vist nedenfor i en aminosyresekvens af et enzym (enzym, der skal muteres), der hydrolyserer en a-l,6-glycosidforbindelse, hvor gruppen består af en aminosyre i substratbindingsstedet af enzymet og involveret i binding af substratet, hvilket svarer til en aminosyre 476-positionen, en aminosyre tilsvarende en aminosyre ved 292-positionen, en aminosyre tilsvarende en aminosyre ved 371-positionen, en aminosyre tilsvarende en aminosyre ved 406-positionen, en aminosyre tilsvarende en aminosyre ved 407-positionen, en aminosyre tilsvarende en aminosyre ved 437-positionen, en aminosyre tilsvarende en aminosyre ved 465-positionen, en aminosyre tilsvarende en aminosyre ved 475-positionen, en aminosyre tilsvarende en aminosyre ved 525-positionen, en aminosyre tilsvarende en aminosyre ved 526-positionen, en aminosyre tilsvarende en aminosyre ved 580-positionen og en aminosyre tilsvarende en aminosyre ved 582-positionen af aminosyresekvensen fremsat i SEQ ID NO: 2 er substitueret med en anden aminosyre eller slettet, hvor aminosyresekvensen af enzymet, der skal muteres, er aminosyresekvensen fremsat i SEQ ID NO: 13 og hvor det muterede enzym er mindre end 10 % forskelligt fra udgangsenzymet i forhold til hele aminosyresekvensen.
2. Det muterede enzym ifølge krav 1, hvor den substituerede eller slettede aminosyre er en eller to eller flere aminosyrer valgt fra gruppen bestående af en aminosyre tilsvarende en aminosyre ved 476-positionen, en aminosyre tilsvarende en aminosyre ved 292-positionen, en aminosyre tilsvarende en aminosyre ved 371-positionen, en aminosyre tilsvarende en aminosyre ved 407-positionen, en aminosyre tilsvarende en aminosyre ved 475-positionen, og en aminosyre tilsvarende en aminosyre ved 582-positionen af aminosyresekvensen fremsat i SEQ ID NO: 2.
3. Gen, der koder for det muterede enzym ifølge krav 1 eller 2.
4. Rekombinant DNA, der inkluderer genet ifølge krav 3.
5. Mikroorganisme, der bærer det rekombinante DNA ifølge krav 4.
DK13179288.9T 2006-08-04 2007-06-28 Fremgangsmåde til at designe muteret enzym, fremgangsmåde til fremstilling af samme og muteret enzym DK2662445T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2006213490 2006-08-04
EP07767790.4A EP2058394B1 (en) 2006-08-04 2007-06-28 Method for designing mutated enzyme, method for preparing the same and mutated enzyme

Publications (1)

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DK2662445T3 true DK2662445T3 (da) 2017-12-04

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DK07767790.4T DK2058394T3 (da) 2006-08-04 2007-06-28 Fremgangsmåde til design af muteret enzym, fremgangsmåde til fremstilling deraf og muteret enzym

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US (1) US20090280553A1 (da)
EP (2) EP2662445B1 (da)
JP (1) JP5178517B2 (da)
CN (1) CN101528923B (da)
DK (2) DK2662445T3 (da)
WO (1) WO2008015861A1 (da)

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DE102009029194A1 (de) 2009-09-04 2011-04-07 Kimberly-Clark Worldwide, Inc., Neenah Abtrennung gefärbter Stoffe aus wasserhaltigen Flüssigkeiten
US9283127B2 (en) 2012-03-30 2016-03-15 Kimberly-Clark Worldwide, Inc. Absorbent articles with decolorizing structures
US9237975B2 (en) 2013-09-27 2016-01-19 Kimberly-Clark Worldwide, Inc. Absorbent article with side barriers and decolorizing agents
CN103789285B (zh) * 2014-02-19 2016-01-20 江南大学 一种热稳定性提高的淀粉酶突变体及其构建方法和应用
JP6585078B2 (ja) 2014-12-05 2019-10-02 合同酒精株式会社 耐熱性イソアミラーゼ
WO2023225459A2 (en) 2022-05-14 2023-11-23 Novozymes A/S Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections
CN114262710B (zh) * 2021-12-31 2023-10-31 西南大学 水稻胞间连丝基因及其突变基因、编码的蛋白和应用

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JP3173849B2 (ja) 1992-02-26 2001-06-04 天野エンザイム株式会社 新規な枝切り酵素及びその製造法
ATE183236T1 (de) * 1992-12-28 1999-08-15 Genencor Int Pullulanase, mikroorganismen die sie produzieren, verfahren zur herstellung und ihre anwendung
JP3372652B2 (ja) 1994-04-26 2003-02-04 天野エンザイム株式会社 炊飯用酵素組成物、それを用いた米飯類の製造法及び製品
AU2980199A (en) * 1998-03-04 1999-09-20 Genencor International, Inc. Modified forms of pullulanase
US6265197B1 (en) * 1998-07-02 2001-07-24 Novozymes A/S Krogshoejvej Starch debranching enzymes
MXPA00012491A (es) * 1998-07-02 2002-07-02 Novozymes As Enzimas desramificantes del almidon.
WO2001051620A2 (en) * 2000-01-12 2001-07-19 Novozymes A/S Pullulanase variants and methods for preparing such variants with predetermined properties
JP2006174841A (ja) * 2006-01-16 2006-07-06 Kao Corp 変異プルラナーゼ

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EP2058394A1 (en) 2009-05-13
CN101528923A (zh) 2009-09-09
WO2008015861A1 (en) 2008-02-07
EP2058394A4 (en) 2009-09-30
EP2662445A2 (en) 2013-11-13
EP2058394B1 (en) 2014-11-12
EP2662445B1 (en) 2017-10-25
JPWO2008015861A1 (ja) 2009-12-17
EP2662445A3 (en) 2014-01-22
US20090280553A1 (en) 2009-11-12
DK2058394T3 (da) 2015-02-02
JP5178517B2 (ja) 2013-04-10
CN101528923B (zh) 2012-03-21

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