DK2290059T3 - Lipolytiske enzymvarianter - Google Patents

Lipolytiske enzymvarianter Download PDF

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Publication number
DK2290059T3
DK2290059T3 DK10182620.4T DK10182620T DK2290059T3 DK 2290059 T3 DK2290059 T3 DK 2290059T3 DK 10182620 T DK10182620 T DK 10182620T DK 2290059 T3 DK2290059 T3 DK 2290059T3
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Denmark
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lipase
lipolytic enzyme
enzyme
activity
amino acid
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DK10182620.4T
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English (en)
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Kirsten Bojsen
Allan Svendsen
Claus Crone Fuglsang
Shamkant Anant Patkar
Kim Borch
Jesper Vind
Andreas Petri
Sanne Schroeder Glad
Gitte Budolfsen
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Novozymes As
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01003—Triacylglycerol lipase (3.1.1.3)
    • 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
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B3/00—Refining fats or fatty oils
    • C11B3/003—Refining fats or fatty oils by enzymes or microorganisms, living or dead
    • 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
    • C11D3/38627—Preparations containing enzymes, e.g. protease or amylase containing lipase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
    • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
    • C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
    • C12H1/04—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material
    • C12H1/0408—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material with the aid of inorganic added material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
    • C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
    • C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
    • C12H1/04—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material
    • C12H1/0416—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material with the aid of organic added material
    • C12H1/0424—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material with the aid of organic added material with the aid of a polymer
    • 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)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C12N9/20—Triglyceride splitting, e.g. by means of lipase
    • 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
    • C12P9/00—Preparation of organic compounds containing a metal or atom other than H, N, C, O, S or halogen
    • 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)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Inorganic Chemistry (AREA)
  • Detergent Compositions (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Seasonings (AREA)
  • Bakery Products And Manufacturing Methods Therefor (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Fats And Perfumes (AREA)
  • Chemical Or Physical Treatment Of Fibers (AREA)
  • General Preparation And Processing Of Foods (AREA)
  • Dairy Products (AREA)

Claims (16)

1. Lipolytisk enzym, som: a) er et polypeptid med en aminosyresekvens, der udviser mindst 80% homologi med et lipolytisk referenceenzym, hvor det lipolytiske referenceenzym er lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1); b) i sammenligning med det lipolytiske referenceenzym omfatter en aminosyre-ændring, der er en substitution svarende til G266A,D,L,S,V,W,E i lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1); og c) har en øget aktivitet på en esterbinding i et substrat i sammenligning med det lipolytiske referenceenzym, hvor aktiviteten er phospholipaseaktivitet.
2. Lipolytisk enzym ifølge krav 1, som endvidere omfatter mindst én yderligere aminosyre-ændring, der er en substitution, en deletion eller en insertion svarende til en hvilken som helst af positionerne 22, 56-59, 61, 64, 83, 85, 91, 94, 249, 255 eller 259, fortrinsvis S83T, G91A, N94D, D96S,W,F,G, Q249R eller L259N,R,S,M,Q, og/eller en substitution svarende til D62A,G,V, K98D, E99K, P256T, G263A og/eller I265T,G,V.
3. Lipolytisk enzym ifølge krav 1-2, som omfatter en aminosyreændring i en position svarende til R81, R84, S85, G263, L264, I265, T267 eller L269 i lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1), fortrinsvis en substitution svarende til G263A,E,Q,R; L264A,C,P,Q; I265L,N,T; eller T267A,Q eller L269N.
4. Lipolytisk enzym ifølge et hvilket som helst af kravene 1-3, som omfatter en peptidforlængelse i den C-terminale ende, hvilken peptidforlængelse fortrinsvis omfatter 1-5 amino-syrerester, hvor den første fortrinsvis er A, P eller D, den anden (hvis den er til stede) fortrinsvis er V, G eller R, den tredje (hvis den er til stede) fortrinsvis er V, G eller R, den fjerde (hvis den er til stede) fortrinsvis er F, og den femte (hvis den er til stede) fortrinsvis er S.
5. Lipolytisk enzym ifølge et hvilket som helst af kravene 1-4, som omfatter en ændring i lågregionen, der er en substitution af en negativt ladet aminosyrerest med en neutral eller positivt ladet aminosyrerest eller en substitution af en neutral aminosyrerest med en positivt ladet aminosyrerest.
6. Lipolytisk enzym ifølge krav 5, som omfatter en ændring i lågregionen i en position svarende til position G91, D96 og/eller E99 i lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1), fortrinsvis en substitution, som er G91A, D96S,W,F eller E99K.
7. Lipolytisk enzym ifølge krav 1, som er en variant af lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1), hvilken variant omfatter ændringerne E1E,D,A +G91G,A,S,T +N94N,D +D96D,G,F,W +E99E.K +G225G,R,K +G263Q.N +L264L,A,V +I265I.T.S +G266A,V,S,D,E +T267T,A,V +L269L,I,N,Q.
8. Lipolytisk enzym ifølge krav 7, som endvidere omfatter SPIRR som en peptidforlængelse i den N-terminale ende og/eller AGGF eller AGGFS som en peptidforlængelse i den C-terminale ende.
9. Lipolytisk enzym ifølge krav 1, som er en variant af lipasen fra Humicola lanuginosa (SP400, som er vist i figur 1), hvor varianten er valgt blandt:
10. DNA-sekvens, som koder for det lipolytiske enzym ifølge et hvilket som helst af kravene 1-9.
11. Vektor, som omfatter DNA-sekvensen ifølge krav 10.
12. Transformeret værtscelle, som huser DNA-sekvensen ifølge krav 10 eller vektoren ifølge krav 11.
13. Fremgangsmåde til fremstilling af det lipolytiske enzym ifølge et hvilket som helst af kravene 1-9, hvilken fremgangsmåde omfatter a) dyrkning af cellen ifølge krav 12 for at udtrykke og fortrinsvis udskille det lipolytiske enzym og b) indvinding af det lipolytiske enzym.
14. Fremgangsmåde til fremstilling af en dej eller et bagt produkt fremstillet af dejen, hvilken fremgangsmåde omfatter tilsætning af det lipolytiske enzym ifølge et hvilket som helst af kravene 1-9 til dejen.
15. Fremgangsmåde til reduktion af indholdet af phospholipid i en spiseolie, hvilken fremgangsmåde omfatter behandling af olien med det lipolytiske enzym ifølge et hvilket som helst af kravene 1-9 for at hydrolysere en væsentlig del af phospholipidet og fraseparation af en vandig fase, som indeholder det hydrolyserede phospholipid, fra olien.
16. Detergentsammensætning, som omfatter et overfladeaktivt middel og det lipolytiske enzym ifølge et hvilket som helst af kravene 1-9.
DK10182620.4T 1998-11-27 1999-11-29 Lipolytiske enzymvarianter DK2290059T3 (da)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DKPA199801572 1998-11-27
DKPA199900391 1999-03-22
DKPA199901481 1999-10-15
EP09165405A EP2113563A3 (en) 1998-11-27 1999-11-29 Lipolytic enzyme variants

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DK2290059T3 true DK2290059T3 (da) 2016-02-22

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JP (3) JP4723087B2 (da)
CN (1) CN100374556C (da)
AT (1) ATE441704T1 (da)
AU (2) AU785464B2 (da)
BR (1) BRPI9915711B1 (da)
CA (2) CA2353379C (da)
DE (1) DE69941364D1 (da)
DK (1) DK2290059T3 (da)
ES (1) ES2328429T3 (da)
TR (1) TR200101497T2 (da)

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AU2007229338B2 (en) 2011-06-02
EP2113563A3 (en) 2010-01-13
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EP2290058A1 (en) 2011-03-02
CA2715086A1 (en) 2000-06-08
CN100374556C (zh) 2008-03-12
ATE441704T1 (de) 2009-09-15
JP2014138591A (ja) 2014-07-31
AU1376300A (en) 2000-06-19
CA2353379C (en) 2011-01-04
EP2290059B1 (en) 2015-11-25
EP2298873A1 (en) 2011-03-23
CA2353379A1 (en) 2000-06-08
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BR9915711A (pt) 2001-08-21
DE69941364D1 (de) 2009-10-15
EP2290059A1 (en) 2011-03-02
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EP2236602A1 (en) 2010-10-06
EP2302043A2 (en) 2011-03-30
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JP2003524386A (ja) 2003-08-19
JP5600055B2 (ja) 2014-10-01
JP4723087B2 (ja) 2011-07-13
TR200101497T2 (tr) 2001-11-21
AU785464B2 (en) 2007-07-26
CN1331742A (zh) 2002-01-16
JP2011083285A (ja) 2011-04-28
ES2328429T3 (es) 2009-11-12
EP2302043A3 (en) 2011-07-20
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BRPI9915711B1 (pt) 2016-01-05
AU2007229338A1 (en) 2007-11-08

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