DK3061817T3 - Alkaliske proteasevarianter - Google Patents

Alkaliske proteasevarianter Download PDF

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Publication number
DK3061817T3
DK3061817T3 DK16163320.1T DK16163320T DK3061817T3 DK 3061817 T3 DK3061817 T3 DK 3061817T3 DK 16163320 T DK16163320 T DK 16163320T DK 3061817 T3 DK3061817 T3 DK 3061817T3
Authority
DK
Denmark
Prior art keywords
amino acid
dna
oligonucleotide
artificial
pcr primer
Prior art date
Application number
DK16163320.1T
Other languages
English (en)
Inventor
Masatoshi Tohata
Mitsuyoshi Okuda
Tsuyoshi Sato
Keiji Endo
Katsuhisa Saeki
Original Assignee
Kao Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP2010093307A external-priority patent/JP2010273672A/ja
Priority claimed from JP2010093339A external-priority patent/JP2010273673A/ja
Application filed by Kao Corp filed Critical Kao Corp
Application granted granted Critical
Publication of DK3061817T3 publication Critical patent/DK3061817T3/da

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Classifications

    • 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
    • C11D3/38618—Protease or amylase in liquid compositions only
    • 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
    • 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
    • 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)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus

Claims (11)

1. Alkalisk proteasevariant, hvor den alkaliske proteasevariant (i) består af aminosyresekvensen ifølge SEQ ID NO: 2, hvor aminosyreresten ved position 83 i aminosyresekvensen ifølge SEQ ID NO: 2 er substitueret med følgende aminosyrerester: alanin, serin eller cystein, eller (ii) har en identitet på 90% eller mere med SEQ ID NO: 2, hvor aminosyreresten ved positionen svarende til position 83 i aminosyresekvensen ifølge SEQ ID NO: 2 er substitueret med følgende aminosyrerester: alanin, serin eller cystein, og hvor den alkaliske proteasevariant udviser aktivitet i et alkalisk pH område.
2. Alkalisk proteasevariant ifølge krav 1, hvor yderligere én eller flere aminosyrerester med positioner valgt blandt (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 204, (h) position 319 og (i) position 337 i aminosyresekvensen ifølge SEQ ID NO: 2, eller ved positioner svarende dertil, er substituerede med følgende aminosyrerester: (a) tryptophan, leucin, valin, isoleucin, methionin, tyrosin, glutamin, lysin, threonin, phenylalanin, arginin, serin, cystein, alanin eller histidin; (b) glutaminsyre, methionin, aspartansyre, valin, glutamin, arginin, cystein, tryptophan, alanin eller phenylalanin; (c) methionin, glutaminsyre, arginin, valin, lysin, phenylalanin, tyrosin, isoleucin, histidin, aspartansyre eller cystein; (d) tryptophan; (e) histidin, tryptophan, serin eller leucin; (f) alanin, glutaminsyre, glutamin, serin, cystein, glycin, histidin, lysin, arginin, methionin eller asparagin; (g) glutaminsyre, aspartansyre, cystein, valin, threonin, prolin, histidin, isoleucin, tryptophan, serin, asparagin, lysin eller arginin; (h) tryptophan, valin, threonin, leucin, isoleucin, cystein, glutaminsyre, lysin, tyrosin, arginin, phenylalanin, glutamin, methionin, prolin, aspartansyre, asparagin, histidin eller serin; og (i) arginin, glycin, serin, lysin, glutamin, threonin, histidin, alanin, cystein eller valin.
3. Gen, som koder for den alkaliske proteasevariant ifølge krav 1 eller 2.
4. Rekombinant vektor omfattende genet ifølge krav 3.
5. Ikke-human transformant, omfattende den rekombinante vektor ifølge krav 4.
6. Den ikke-humane transformant ifølge krav 5, hvis vært er en mikroorganisme.
7. Vaskemiddelsammensætning omfattende den alkaliske proteasevariant ifølge krav 1 eller 2.
8. Vaskemiddelsammensætning ifølge krav 7, som omfatter et anionisk overfladeaktivt stof.
9. Fremgangsmåde til at fremme stabiliteten af en alkalisk protease i et flydende vaskemiddel, hvilken fremgangsmåde omfatter (i) i en alkalisk protease bestående af aminosyresekvensen ifølge SEQ ID NO: 2, at substituere aminosyreresten ved position 83 i aminosyresekvensen ifølge SEQ ID NO: 2 med følgende aminosyrerester: alanin, serin eller cystein; eller (ii) i en alkalisk protease med en identitet på 90% eller mere med SEQ ID NO: 2, at substituere aminosyreresten ved positionen svarende til position 83 i aminosyresekvensen ifølge SEQ ID NO: 2 med følgende aminosyrerester: alanin, serin eller cystein, hvor den fremstillede alkaliske proteasevariant udviser aktivitet i et alkalisk pH område.
10. Fremgangsmåde ifølge krav 9, hvor, i den alkaliske protease, én eller flere aminosyrerester yderligere er substituerede ved positioner valgt blandt (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 204, (h) position 319, and (i) position 337 i aminosyresekvensen ifølge SEQ ID NO: 2, eller ved positioner svarende dertil, med følgende aminosyrerester: (a) tryptophan, leucin, valin, isoleucin, methionin, tyrosin, glutamin, lysin, threonin, phenylalanin, arginin, serin, cystein, alanin eller histidin; (b) glutaminsyre, methionin, aspartansyre, valin, glutamin, arginin, cystein, tryptophan, alanin eller phenylalanin; (c) methionin, glutaminsyre, arginin, valin, lysin, phenylalanin, tyrosin, isoleucin, histidin, aspartansyre eller cystein; (d) tryptophan; (e) histidin, tryptophan, serin eller leucin; (f) alanin, glutaminsyre, glutamin, serin, cystein, glycin, histidin, lysin, arginin, methionin eller asparagin; (g) glutaminsyre, aspartansyre, cystein, valin, threonin, prolin, histidin, isoleucin, tryptophan, serin, asparagin, lysin eller arginin; (h) tryptophan, valin, threonin, leucin, isoleucin, cystein, glutaminsyre, lysin, tyrosin, arginin, phenylalanin, glutamin, methionin, prolin, aspartansyre, asparagin, histidin eller serin; og (i) arginin, glycin, serin, lysin, glutamin, threonin, histidin, alanin, cystein eller valin.
11. Fremgangsmåde til fremstilling af den alkaliske proteasevariant ifølge krav 1 eller 2, hvilken fremgangsmåde omfatter dyrkning af transformanten ifølge krav 5.
DK16163320.1T 2009-04-30 2010-04-26 Alkaliske proteasevarianter DK3061817T3 (da)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
JP2009110792 2009-04-30
JP2009110793 2009-04-30
JP2010093307A JP2010273672A (ja) 2009-04-30 2010-04-14 アルカリプロテアーゼ変異体
JP2010093339A JP2010273673A (ja) 2009-04-30 2010-04-14 アルカリプロテアーゼ変異体
EP10719131A EP2424981A2 (en) 2009-04-30 2010-04-26 Alkaline protease variants

Publications (1)

Publication Number Publication Date
DK3061817T3 true DK3061817T3 (da) 2019-03-04

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DK16163320.1T DK3061817T3 (da) 2009-04-30 2010-04-26 Alkaliske proteasevarianter

Country Status (5)

Country Link
US (1) US8778650B2 (da)
EP (2) EP3061817B1 (da)
CN (1) CN102421893B (da)
DK (1) DK3061817T3 (da)
WO (1) WO2010126156A2 (da)

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Publication number Priority date Publication date Assignee Title
CN103275958B (zh) * 2011-08-01 2014-07-16 南京工业大学 一种耐有机溶剂碱性蛋白酶
CN102703482B (zh) * 2011-08-01 2014-08-06 南京工业大学 一种耐有机溶剂碱性蛋白酶
JP6067409B2 (ja) 2012-04-10 2017-01-25 花王株式会社 アルカリプロテアーゼの溶解性向上方法
CN103409398B (zh) * 2013-09-03 2014-12-24 广州立白企业集团有限公司 一种蛋白酶及突变体及其应用
JP2018521653A (ja) * 2015-07-29 2018-08-09 シージェイ チェルジェダン コーポレイション 転換活性が高められたヘキスロン酸ｃ４−エピメラーゼ変異体及びこれを用いたｄ−タガトースの製造方法
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Also Published As

Publication number Publication date
EP3061817A1 (en) 2016-08-31
WO2010126156A2 (en) 2010-11-04
US20120058928A1 (en) 2012-03-08
WO2010126156A3 (en) 2011-01-06
CN102421893B (zh) 2014-10-29
US8778650B2 (en) 2014-07-15
EP3061817B1 (en) 2018-12-12
EP2424981A2 (en) 2012-03-07
CN102421893A (zh) 2012-04-18

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