DK3097192T3 - Pullulanasevarianter og polynukleotider, som koder for dem - Google Patents
Pullulanasevarianter og polynukleotider, som koder for dem Download PDFInfo
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- DK3097192T3 DK3097192T3 DK15703235.0T DK15703235T DK3097192T3 DK 3097192 T3 DK3097192 T3 DK 3097192T3 DK 15703235 T DK15703235 T DK 15703235T DK 3097192 T3 DK3097192 T3 DK 3097192T3
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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
- C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
- C12N9/2457—Pullulanase (3.2.1.41)
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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
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- 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
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- 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
- C12P19/16—Preparation of compounds containing saccharide radicals produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
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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)
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- 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
- C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
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- 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
Claims (8)
1. Pullulanasevariant med forøget termoaktivitet, som omfatter en substitution i mindst en position svarende til position 393 i polypeptidet ifølge SEQ ID NO: 3, hvor varianten har pullulanaseaktivitet, og a) hvor varianten udviser mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100% sekvensidentitet med polypeptidet ifølge SEQ ID NO: 3, og hvor varianten har forøget termoaktivitet i sammenligning med pullulanasen ifølge SEQ ID NO: 3, navnlig hvor varianten har mindst 60% relativ aktivitet som målt ved 70°C i forhold til aktiviteten ved 65°C, fortrinsvis mindst 65%, mere fortrinsvis mindst 70%, mere fortrinsvis mindst 60%, mere fortrinsvis mindst 75%, mere fortrinsvis mindst 80%, mere fortrinsvis mindst 90%, mere fortrinsvis mindst 100%; eller b) hvor varianten udviser mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100% sekvensidentitet med polypeptidet ifølge SEQ ID NO: 6, og hvor varianten har forøget termoaktivitet i sammenligning med pullulanasen ifølge SEQ ID NO: 6, navnlig hvor varianten har mindst 30% relativ aktivitet som målt ved 70°C i forhold til aktiviteten ved 65°C, fortrinsvis mindst 40%, mere fortrinsvis mindst 50%, mere fortrinsvis mindst 60%, mere fortrinsvis mindst 70%, mere fortrinsvis mindst 80%, mere fortrinsvis mindst 90%, mere fortrinsvis mindst 100%; eller c) hvor varianten udviser mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100% sekvensidentitet med polypeptidet ifølge SEQ ID NO: 9, og hvor varianten har forøget termoaktivitet i sammenligning med pullulanasen ifølge SEQ ID NO: 9, navnlig hvor varianten har mindst 30% relativ aktivitet som målt ved 70°C i forhold til aktiviteten ved 65°C, fortrinsvis mindst 40%, mere fortrinsvis mindst 50%, mere fortrinsvis mindst 60%, mere fortrinsvis mindst 70%, mere fortrinsvis mindst 80%, mere fortrinsvis mindst 90%, mere fortrinsvis mindst 100%; eller d) hvor varianten udviser mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100% sekvensidentitet med polypeptidet ifølge SEQ ID NO: 16, og hvor varianten har forøget termoaktivitet i sammenligning med pullulanasen ifølge SEQ ID NO: 16, navnlig hvor varianten har mindst 30% relativ aktivitet som målt ved 70°C i forhold til aktiviteten ved 65°C, fortrinsvis mindst 40%, mere fortrinsvis mindst 50%, mere fortrinsvis mindst 60%, mere fortrinsvis mindst 70%, mere fortrinsvis mindst 80%, mere fortrinsvis mindst 90%, mere fortrinsvis mindst 100%; eller e) hvor varianten udviser mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100% sekvensidentitet med polypeptidet ifølge SEQ ID NO: 17, og hvor varianten har forøget termoaktivitet i sammenligning med pullulanasen ifølge SEQ ID NO: 17, navnlig hvor varianten har mindst 30% relativ aktivitet som målt ved 70°C i forhold til aktiviteten ved 65°C, fortrinsvis mindst 40%, mere fortrinsvis mindst 50%, mere fortrinsvis mindst 60%, mere fortrinsvis mindst 70%, mere fortrinsvis mindst 80%, mere fortrinsvis mindst 90%, mere fortrinsvis mindst 100%; og, endvidere, hvor substitutionen i position 393 er 393A.
2. Variant ifølge et hvilket som helst af de foregående krav, hvor varianten omfatter mindst én af følgende kombinationer af substitutioner under anvendelse af SEQ ID NO: 3 til nummerering: N393A + T631S + S632C; N393A + V486C + T492S,A + T631S + S632C; N393A + G624S + S632C; N393A + N610R + T631S + S632C; N393A + G624S + T631S + S632C; N393A + N610R + G624S + T631S + S632C; N393A + V486C + T492S,A + G624S + T631S + S632C; N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; N368G + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + V486C + T492S,A + N610R + G624S + T631S + S632C + E699R; N346S + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + F456A + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + T492S,A + N610R + G624S + T631S + S632C; N368G + N393A + T492S,A + N610R + G624S + T631S + S632C; A345P + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; N368G + K370S + I373L + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; 1381V + Q385E + Q387L + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; 1381V + N382T + Q385E + Q387L + N393A + V486C + T492S,A + N610R + G624S + T631S + S632C; A345P + N368G + N393A + T492S,A + N610R + G624S + T631S + S632C; N368G + 1381V + Q385E + Q387L + N393A + T492S,A + N610R + G624S + T631S + S632C; A345P + N368G + 1381V + Q385E + Q387L + N393A + T492S,A + N610R + G624S + T631S + S632C; A345P + N368G + 1381V + Q385E + Q387L + N393A + T492S,A + N610R + G624S + T631S + S632C + V665I; N393A + T430R + Q431E + L432F + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + Q431E + L432F + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + I429V + Q431E + V486C + T492S,A + N610R + G624S + T631S + S632C; N393A + I429V + T430R + Q431E + L432F + V486C + T492S,A + N610R + G624S + T631S + S632C; N368G + N393A + A492S.A; N368G + N393A; N393A+ N610R; N368G + N393A+ N610R; N368G + N393A + T492S,A + N610R + G624S; N368G + N393A + T492S,A + N610R + G624S + T631S + S632C + Q431E + L432F; N368G + N393A + N610R + G624S + T631S + S632C; N368G + N393A + T492S,A + N610R + G624S + T631S + S632C; N368G + N393A+ N610R + G624S + T631S + S632C + Q431E + L432F.
3. Sammensætning, som omfatter pullulanasevarianten ifølge et hvilket som helst af kravene 1-2.
4. Anvendelse af en pullulanasevariant ifølge et hvilket som helst af kravene 1-2 til fremstilling af en sirup og/eller et fermenteringsprodukt, f.eks. ethanol, ud fra et stivelsesholdigt materiale.
5. Fremgangsmåde til fremstilling af et fermenteringsprodukt ud fra stivelsesholdigt materiale, hvilken fremgangsmåde omfatter følgende trin: (a) at gøre stivelsesholdigt materiale flydende i nærværelse af en alfa-amylase; (b) at forsukre materialet, som er gjort flydende, i nærværelse af en glucoamylase; og (c) at fermentere med en fermenterende organisme; hvor trin (a) og/eller trin (b) udføres i nærværelse af en pullulanasevariant ifølge et hvilket som helst af kravene 1-2.
6. Fremgangsmåde til fremstilling af et fermenteringsprodukt ud fra stivelsesholdigt materiale, hvilken fremgangsmåde omfatter følgende trin: (a) at forsukre stivelsesholdigt materiale ved en temperatur, som er lavere end den laveste gelatineringstemperatur for det stivelsesholdige materiale; og (b) at fermentere med en fermenterende organisme, hvor trin (a) udføres ved anvendelse af mindst en glucoamylase og en pullulanasevariant ifølge et hvilket som helst af kravene 1-2.
7. Polynukleotid, som koder for pullulanasevarianten ifølge et hvilket som helst af kravene 1-2.
8. Værtscelle, som omfatter polynukleotidet ifølge krav 7.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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EP14152110 | 2014-01-22 | ||
EP14195691 | 2014-12-01 | ||
PCT/EP2015/051145 WO2015110473A2 (en) | 2014-01-22 | 2015-01-21 | Pullulanase variants and polynucleotides encoding same |
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DK3097192T3 true DK3097192T3 (da) | 2018-11-19 |
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DK18182098.6T DK3415624T3 (da) | 2014-01-22 | 2015-01-21 | Pullulanasevarianter og polynukleotider, som koder for dem |
DK15703235.0T DK3097192T3 (da) | 2014-01-22 | 2015-01-21 | Pullulanasevarianter og polynukleotider, som koder for dem |
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US (3) | US10030237B2 (da) |
EP (2) | EP3415624B1 (da) |
CN (1) | CN105960457B (da) |
CA (1) | CA2932239C (da) |
DK (2) | DK3415624T3 (da) |
HU (1) | HUE056192T2 (da) |
WO (1) | WO2015110473A2 (da) |
Families Citing this family (12)
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CA2932239C (en) | 2014-01-22 | 2023-03-14 | Novozymes A/S | Pullulanase variants and polynucleotides encoding same |
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WO2016087327A1 (en) | 2014-12-01 | 2016-06-09 | Novozymes A/S | Polypeptides having pullulanase activity comprising the x25, x45 and cbm41 domains |
CA2989314A1 (en) * | 2015-07-21 | 2017-01-26 | Novozymes A/S | Polypeptides having pullulanase activity suitable for use in liquefaction |
CN108384770B (zh) * | 2018-03-01 | 2019-11-22 | 江南大学 | 一种降低环糊精对普鲁兰酶抑制作用的方法 |
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- 2015-01-21 CA CA2932239A patent/CA2932239C/en active Active
- 2015-01-21 EP EP18182098.6A patent/EP3415624B1/en active Active
- 2015-01-21 US US15/112,096 patent/US10030237B2/en active Active
- 2015-01-21 WO PCT/EP2015/051145 patent/WO2015110473A2/en active Application Filing
- 2015-01-21 DK DK18182098.6T patent/DK3415624T3/da active
- 2015-01-21 DK DK15703235.0T patent/DK3097192T3/da active
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US10030237B2 (en) | 2018-07-24 |
US10640755B2 (en) | 2020-05-05 |
US20170044511A1 (en) | 2017-02-16 |
US20200248157A1 (en) | 2020-08-06 |
US11136568B2 (en) | 2021-10-05 |
EP3097192B1 (en) | 2018-07-25 |
WO2015110473A2 (en) | 2015-07-30 |
EP3415624B1 (en) | 2021-08-04 |
DK3415624T3 (da) | 2021-11-08 |
CA2932239C (en) | 2023-03-14 |
EP3415624A1 (en) | 2018-12-19 |
CN105960457A (zh) | 2016-09-21 |
WO2015110473A3 (en) | 2015-09-17 |
CN105960457B (zh) | 2021-09-03 |
US20180298364A1 (en) | 2018-10-18 |
EP3097192A2 (en) | 2016-11-30 |
HUE056192T2 (hu) | 2022-01-28 |
CA2932239A1 (en) | 2015-07-30 |
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