JP5933710B2 - 配列リストを参照するアミラーゼ変異体を含む洗浄組成物 - Google Patents
配列リストを参照するアミラーゼ変異体を含む洗浄組成物 Download PDFInfo
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- JP5933710B2 JP5933710B2 JP2014519056A JP2014519056A JP5933710B2 JP 5933710 B2 JP5933710 B2 JP 5933710B2 JP 2014519056 A JP2014519056 A JP 2014519056A JP 2014519056 A JP2014519056 A JP 2014519056A JP 5933710 B2 JP5933710 B2 JP 5933710B2
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- FJBHGWADYLMEJG-UHFFFAOYSA-M sodium;3-[[4-[[4-(diethylamino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]cyclohexa-2,5-dien-1-ylidene]methyl]-n-ethylanilino]methyl]benzenesulfonate Chemical compound [Na+].C1=CC(N(CC)CC)=CC=C1C(C=1C=CC(=CC=1)N(CC)CC=1C=C(C=CC=1)S([O-])(=O)=O)=C(C=C1)C=CC1=[N+](CC)CC1=CC=CC(S([O-])(=O)=O)=C1 FJBHGWADYLMEJG-UHFFFAOYSA-M 0.000 description 1
- KVCGISUBCHHTDD-UHFFFAOYSA-M sodium;4-methylbenzenesulfonate Chemical compound [Na+].CC1=CC=C(S([O-])(=O)=O)C=C1 KVCGISUBCHHTDD-UHFFFAOYSA-M 0.000 description 1
- QEKATQBVVAZOAY-UHFFFAOYSA-M sodium;4-propan-2-ylbenzenesulfonate Chemical compound [Na+].CC(C)C1=CC=C(S([O-])(=O)=O)C=C1 QEKATQBVVAZOAY-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000007614 solvation Methods 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 1
- 235000021286 stilbenes Nutrition 0.000 description 1
- 229940115922 streptococcus uberis Drugs 0.000 description 1
- 125000005504 styryl group Chemical group 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 150000003457 sulfones Chemical class 0.000 description 1
- 238000004381 surface treatment Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 108010075550 termamyl Proteins 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000001302 tertiary amino group Chemical group 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 125000005207 tetraalkylammonium group Chemical group 0.000 description 1
- QTTDXDAWQMDLOF-UHFFFAOYSA-J tetrasodium 3-[[4-[[4-[(6-amino-1-hydroxy-3-sulfonatonaphthalen-2-yl)diazenyl]-6-sulfonatonaphthalen-1-yl]diazenyl]naphthalen-1-yl]diazenyl]naphthalene-1,5-disulfonate Chemical compound [Na+].[Na+].[Na+].[Na+].Nc1ccc2c(O)c(N=Nc3ccc(N=Nc4ccc(N=Nc5cc(c6cccc(c6c5)S([O-])(=O)=O)S([O-])(=O)=O)c5ccccc45)c4ccc(cc34)S([O-])(=O)=O)c(cc2c1)S([O-])(=O)=O QTTDXDAWQMDLOF-UHFFFAOYSA-J 0.000 description 1
- 235000019818 tetrasodium diphosphate Nutrition 0.000 description 1
- 239000001577 tetrasodium phosphonato phosphate Substances 0.000 description 1
- JADVWWSKYZXRGX-UHFFFAOYSA-M thioflavine T Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C1=[N+](C)C2=CC=C(C)C=C2S1 JADVWWSKYZXRGX-UHFFFAOYSA-M 0.000 description 1
- 150000003573 thiols Chemical group 0.000 description 1
- 229930192474 thiophene Natural products 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 102000003601 transglutaminase Human genes 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 125000005208 trialkylammonium group Chemical group 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 235000013799 ultramarine blue Nutrition 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 150000003672 ureas Chemical class 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- YKSGNOMLAIJTLT-UHFFFAOYSA-N violanthrone Chemical compound C12=C3C4=CC=C2C2=CC=CC=C2C(=O)C1=CC=C3C1=CC=C2C(=O)C3=CC=CC=C3C3=CC=C4C1=C32 YKSGNOMLAIJTLT-UHFFFAOYSA-N 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
- 239000004711 α-olefin Substances 0.000 description 1
Classifications
-
- 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
-
- 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
- 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)
- C12Y302/01001—Alpha-amylase (3.2.1.1)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Detergent Compositions (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
(a)配列番号1の成熟ポリペプチドの位置G304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、及びG477に対応する2つ以上(複数)の位置での変性を含む、親アルファ−アミラーゼの変異体であって、それぞれの変性が独立して、置換、欠失、又は挿入であり、変異体が配列番号1、2、3、4、5、6、7、8、9、10、11、又は12のいずれかの成熟ポリペプチドと少なくとも80%、又は少なくとも87%であるが、100%未満である配列同一性を有し、変異体がアルファ−アミラーゼ活性を有する、変異体と、
(b)好ましくは0.01〜99.9重量%の量の洗浄補助剤とを含む洗浄組成物を提供する。
及び
(a)配列番号1の成熟ポリペプチドの位置G304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、及びG477に対応する2つ以上(複数)の位置での変性を含む、親アルファ−アミラーゼの変異体であって、それぞれの変性が独立して、置換、欠失、又は挿入であり、変異体が配列番号1、2、3、4、5、6、7、8、9、10、11、又は12のいずれかの成熟ポリペプチドと少なくとも80%、又は少なくとも87%であるが、100%未満である配列同一性を有し、変異体がアルファ−アミラーゼ活性を有する、変異体と、
(b)好ましくは0.01〜99.9重量%の量の洗浄補助剤とを含む、工程と、
(ii)好ましくは40℃以下の温度、より好ましくは30℃以下の温度、最も好ましくは20℃以下の温度の水性洗浄溶液で表面を処理する工程と、(iii)表面をすすぐ工程とを含む。
(i)水及びそのような洗浄組成物を含む水性洗浄溶液を形成する工程と、
(ii)好ましくは40℃以下の温度、より好ましくは30℃以下の温度、最も好ましくは20℃以下の温度の水性洗浄溶液で表面を処理する工程と、
(iii)表面をすすぐ工程とを含む。
(a)配列番号1の成熟ポリペプチドの位置G304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、及びG477に対応する2つ以上(複数)の位置での変性を含む、親アルファ−アミラーゼの変異体であって、それぞれの変性が独立して、置換、欠失、又は挿入であり、変異体が配列番号1、2、3、4、5、6、7、8、9、10、11、又は12のいずれかの成熟ポリペプチドと少なくとも80%、又は少なくとも87%であるが、100%未満である配列同一性を有し、変異体がアルファ−アミラーゼ活性を有する、変異体と、
(b)好ましくは0.01〜99.9重量%の量の洗浄補助剤とを含む洗浄組成物を提供する。
アルファ−アミラーゼ活性:用語「アルファ−アミラーゼ活性」とは、デンプン並びに他の直鎖状及び分枝状1,4−グルコシドオリゴ並びに多糖類の加水分解を触媒する酵素の一群を構成するアルファ−1,4−グルカン−4−グルカノヒドロラーゼ、E.C.3.2.1.1の活性を意味する。
(同一残基×100)/(アラインメントの長さ−アラインメント中のギャップの総数)
(同一デオキシリボヌクレオチド×100)/(アラインメントの長さ−アラインメント中のギャップの総数)。
本発明の目的のため、配列番号1に開示される成熟ポリペプチドは、別のアルファ−アミラーゼの対応するアミノ酸残基を決定するために使用される。別のアルファ−アミラーゼのアミノ酸配列は、配列番号1、配列番号2、配列番号3、配列番号4、配列番号5、配列番号6に開示される成熟ポリペプチドとアライメントされ、アライメントに基づき、配列番号1に開示される成熟ポリペプチドのいずれかのアミノ酸残基に対応するアミノ酸位置番号が、EMBOSSパッケージ(EMBOSS:The European Molecular Biology Open Software Suite,Rice et al.,2000,Trends Genet.16:276〜277)、好ましくはバージョン3.0.0以降のNeedleプログラムにおいて実施されるようなNeedleman−Wunschアルゴリズム(Needleman and Wunsch,1970,J.Mol.Biol.48:443〜453)を用いて決定される。
「Tyr167Gly+Arg170Gly」、「Tyr167Gly+Arg170Ala」、「Tyr167Ala+Arg170Gly」、及び「Tyr167Ala+Arg170Ala」を示す。
親アルファ−アミラーゼは、配列番号1の成熟ポリペプチドと少なくとも80%の配列同一性を有するポリペプチドであり得る。
アルファ−アミラーゼ活性を有する変異体を得るための方法は、(a)任意の配列番号1、配列番号2、配列番号3、配列番号4、配列番号5、配列番号6、配列番号7、配列番号8、配列番号9、配列番号10、配列番号11、又は配列番号12の成熟ポリペプチドの位置195、206、243に対応する位置での1つ以上の変性に加え、親アルファ−アミラーゼの中に、位置140、181、189、134、260、262、284、304、347、439、469、476、及び477に対応する2つ以上(複数)の位置で変性を導入する工程であって、番付は配列番号1に従い、変異体はアルファ−アミラーゼ活性を有する工程と、(b)変異体を回収する工程とを含み得る。
本発明に有用な親アルファ−アミラーゼの変異体は、配列番号1の成熟ポリペプチドの位置G304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、及びG477に対応する2つ以上(複数)の位置で変性を含み、それぞれの変性は、独立して、置換、挿入、又は欠失(好ましくは置換)であり、変異体はアルファ−アミラーゼ活性を有する。これによって、親アルファ−アミラーゼと比較して、又は配列番号1、2、3、4、5、6、7、8、9、10、11、又は12のアルファ−アミラーゼと比較して、低温での改善された洗浄性能を有する変異体が提供される。
W140Y+N195F+V206Y+Y243F+E260G+G477E、
W140Y+N195F+V206Y+Y243F+E260T+W284D、
W140Y+N195F+V206Y+Y243F+W284D、
G109A+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G、
N195F+V206Y+Y243F+E260K+W284D、
D134E+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+W189G+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+S303G、
W140Y+W189T+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+W284D、
Y100I+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+G337N、
W140Y+N195F+V206Y+Y243F+E260G+W439R、
G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
N195F+V206Y+Y243F+E260G+W284Dからなる群から選択される、配列番号1のポリペプチドの位置に対応する位置での置換を含む変異体に関する。
W140Y+N195F+V206Y+Y243F+E260G+G477E、
W140Y+N195F+V206Y+Y243F+E260T+W284D、
W140Y+N195F+V206Y+Y243F+W284D、
G109A+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G、
N195F+V206Y+Y243F+E260K+W284D、
D134E+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+W189G+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+S303G、
W140Y+W189T+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+W284D、
Y100I+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+G337N、
W140Y+N195F+V206Y+Y243F+E260G+W439R、
G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
N195F+V206Y+Y243F+E260G+W284Dからなる群から選択される、配列番号1のポリペプチドの位置に対応する位置での置換からなる変異体に関する。
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G477E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260T+W284D、
D183*+G184*+W140Y+N195F+V206Y+Y243F+W284D、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+N195F+V206Y+Y243F+E260K+W284D、
D183*+G184*+D134E+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+W189G+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+S303G、
D183*+G184*+W140Y+W189T+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284D、
D183*+G184*+Y100I+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G337N、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
D183*+G184*+N195F+V206Y+Y243F+E260G+W284Dからなる群から選択される、配列番号1のポリペプチドの位置に対応する位置で変性を含む。
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G477E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260T+W284D、
D183*+G184*+W140Y+N195F+V206Y+Y243F+W284D、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+N195F+V206Y+Y243F+E260K+W284D、
D183*+G184*+D134E+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+W189G+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+S303G、
D183*+G184*+W140Y+W189T+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284D、
D183*+G184*+Y100I+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G337N、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G
D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
D183*+G184*+N195F+V206Y+Y243F+E260G+W284Dからなる群から選択される配列番号1のポリペプチドの位置に対応する位置での変性からなる。
本発明は、本発明の変異体を含む組成物にも関する。好ましくは、組成物はそのような変異体に濃縮される。用語「濃縮」とは、組成物のアルファ−アミラーゼ活性が増加して、例えば濃縮係数が1.1になることを意味する。
オキシスポルム(Fusarium oxysporum)、フザリウム・レチクラツム(Fusarium reticulatum)、フザリウム・ロセウム(Fusarium roseum)、フザリウム・サンブシヌム(Fusarium sambucinum)、フザリウム・サルコクロウム(Fusarium sarcochroum)、フザリウム・スルフレウム(Fusarium sulphureum)、フザリウム・トルロセウム(Fusarium toruloseum)、フザリウム・トリコテシオイデス(Fusarium trichothecioides)、若しくはフザリウム・ベエナツム(Fusarium venenatum)、フミコーラ(Humicola)属、例えばフミコーラ・インソレンス(Humicola insolens)、若しくはフミコーラ・ランギノーサ(Humicola lanuginosa)、又はトリコデルマ(Trichoderma)属、例えばトリコデルマ・ハルジアヌム(Trichoderma harzianum)、トリコデルマ・コニンギイ(Trichoderma koningii)、トリコデルマ・ロンギブラキアタム(Trichoderma longibrachiatum)、トリコデルマ・リーセイ(Trichoderma reesei)、若しくはトリコデルマ・ビリデ(Trichoderma viride)。
本発明は、好ましくは、消費者用又は業務用のエアケア、カーケア、食器洗浄、布地柔軟剤(軟化剤を含む)、洗濯洗浄力、洗濯及びリンス剤及び/又はケア、塗装面洗浄及び/又はトリートメント、並びに他の洗浄のための特許請求される組成物に関する方法及び/又はその使用のための製品に関する。本発明による、上述のアルファ−アミラーゼ変異体は、典型的には、固形、液体、ゲル、及び/又は単位用量の洗浄組成物、例えば、洗濯洗剤組成物又は食器洗浄洗剤組成物中の構成要素であり得る。液体洗濯洗剤組成物が特に好ましい。
(a)米国特許第6,312,936(B1)号、同第5,679,630号、同第4,760,025号、同第7,262,042号及び国際公開第09/021867号に記載のバチルス・レンタス、B.アルカロフィルス(B. alkalophilus)、枯草菌、B.アミロリケファシエンス、バチルス・プミルス及びバチルス・ギブソニィなどのバチルスから誘導されたものなどのサブチリシン(EC 3.4.21.62)。
(b)トリプシン(例えばブタ又はウシ由来)などのトリプシン型又はキモトリプシン型プロテアーゼ(PCT国際公開特許WO 89/06270号に記述されているフサリウムプロテアーゼ、及び同WO 05/052161号及び同WO 05/052146に記述されているCellumonasに由来するキモトリプシンプロテアーゼを含む)。
(c)メタロプロテアーゼ(国際公開第07/044993(A2)号に記述されているBacillus amyloliquefaciensから誘導されたものを含む)。
(a)米国特許第7,141,403B2号の配列番号2のアミノ酸配列に対して少なくとも90%、94%、97%、及び更には99%の同一性の配列を有するバチルス(Bacillus)属のメンバーに内因性の細菌ポリペプチドと、
(b)キシログルカンと非晶質セルロース基質の両方に対して酵素活性を有するグリコシル加水分解酵素であって、GHファミリー5、12、44又は74から選択される、グリコシル加水分解酵素と、
(c)国際公開第WO09/148983号の配列番号3のアミノ酸配列に対して少なくとも90%、94%、97%、及び更には99%の同一性の配列を有するブリコシル加水分解酵素と、
(d)それらの混合物を含む群から選択される。
消費者製品は1つ以上のポリマーを含み得る。例には、カルボキシメチルセルロース、ポリ(ビニル−ピロリドン)、ポリ(エチレングリコール)、ポリ(ビニルアルコール)、ポリ(ビニルピリジン−N−オキシド)、ポリ(ビニルイミダゾール)、ポリアクリル酸塩等のポリカルボン酸塩、マレイン酸/アクリル酸コポリマー、及びメタクリル酸ラウリル/アクリル酸コポリマー、並びに両親媒性ポリマーがある。
好ましくは、両親媒性の洗浄用ポリマーは、以下の一般構造式:ビス((C2H5O)(C2H4O)n)(CH3)−N+−CxH2x−N+−(CH3)−ビス((C2H5O)(C2H4O)n)を有する化合物、又はこれらのサルフェート化若しくはスルホネート化変異体であり、式中、n=20〜30であり、かつx=3〜8である。
A2は、各場合において、1,2−プロピレン、1,2−ブチレン及び1,2−イソブチレンから選択され、好ましくはA2は、1,2−プロピレンである。A3は1,2−プロピレンであり、Rは、各場合において、水素及びC1〜C4アルキル(メチル、エチル、n−プロピル、イソプロピル、n−ブチル、イソブチル及びtert−ブチルなど)から選択され、好ましくはRは水素である。添え字mは、各場合において、0〜約2の値を有し、好ましくはmは0又はおよそ1であり、より好ましくはmは0である。添え字nは、約20〜約50の範囲、好ましくは約22〜約40の範囲、より好ましくは約24〜約30の範囲の平均値を有する。添え字pは、約10〜約50の範囲、好ましくは約11〜約40の範囲、より好ましくは約12〜約30の範囲の平均値を有する。
(I) −[(OCHR1−CHR2)a−O−OC−Ar−CO−]d
(II) −[(OCHR3−CHR4)b−O−OC−sAr−CO−]e
(III) −[(OCHR5−CHR6)c−OR7]f
式中、
a、b、及びcは、1〜200であり、
d、e、及びfは、1〜50であり、
Arは、1,4−置換フェニレンであり、
sArは、位置5をSO3Meにより置換された、1,3−置換フェニレンであり、
Meは、Li、K、Mg/2、Ca/2、Al/3、アンモニウム、モノ−、ジ−、トリ−、若しくはテトラアルキルアンモニウム(アルキル基は、C1〜C18アルキル又はC2〜C10ヒドロキシアルキル)、又はこれらの混合物であり、
R1、R2、R3、R4、R5、及びR6は、独立して、H、又はC1〜C18 n−若しくはイソ−アルキルから選択され;
R7は、直鎖若しくは分枝鎖C1〜C18アルキル、又は直鎖若しくは分枝鎖C2〜C30アルケニル、又は炭素原子を5〜9個有するシクロアルキル基、又はC8〜C30アリール基、又はC6〜C30アリールアルキル基である。
アルファ−アミラーゼ活性は、G7−pNP基質を採用する方法によって決定することができる。G7−pNPは、4,6−エチリデン(G7)−p−ニトロフェニル(G1)−α、D−マルトヘプタオシドの省略形であり、アルファ−アミラーゼ等のエンドアミラーゼで切断することができるブロックオリゴ糖である。切断後、キット中に含まれるアルファ−グルコシダーゼは、加水分解基質を更に消化して、黄色を有する遊離PNP分子を自由にし、ひいては、λ=405nm(400〜420nm)で、可視分光測光で測定することができる。G7−pNP基質及びα−グルコシダーゼを含有するキットは、Roche/Hitachi(カタログ番号No.11876473)によって製造される。
このキットのG7−pNP基質は、22mMの4,6−エチリデン−G7−pNP及び52.4mMのHEPES(2−[4−(2−ヒドロキシエチル)−1−ピペラジニル]−エタンスルホン酸)(pH 7.0)を含有する。
分析されるアミラーゼサンプルは、希釈されるサンプルのpHが7であることを確実にするために、希釈緩衝液中で希釈された。20μlの希釈された酵素サンプルを96ウェルマイクロタイタープレートに移し、80μLの基質作業溶液を添加することにより、アッセイを行った。溶液を混合し、室温で1分間事前インキュベートし、吸収を20秒毎に5分間、OD 405nmで測定する。
洗濯における洗浄性能を評価するために、自動機械応力アッセイ(AMSA)を用いて洗浄実験を行った。AMSAで、大量の少容量酵素−洗剤溶液の洗浄性能を試験することが可能である。AMSAプレートは、試験溶液のための多くのスロット及び洗浄される洗濯サンプルである織物をスロットの全開口部にしっかりと締め付けるリッドを有している。洗浄期間中、プレート、試験溶液、織物及びリッドを、試験溶液を織物と接触させ、規則的及び周期的な振動様式で機械的ストレスを印加するために、精力的に振盪する。更なる説明に関して、国際特許第WO02/42740号、特に23〜24ページの段落「Special method embodiments」を参照されたい。
水(10度dH)、洗剤、例えば以下に記載する5.1g/Lのヨーロッパの液体洗剤、並びに例えば0,0.8及び/又は1.2mgの酵素タンパク質/Lの濃度の本発明の酵素を含む試験溶液を調製する。デンプン(例えばCenter For Testmaterials BV,P.O.Box 120,3133 KT,Vlaardingen,The NetherlandsのCS−28)で染色した布地を添加し、20℃で20分間洗浄する。水道水を流して完全にすすぎ、暗所で乾燥させた後、続いて、染色した布地の光強度又は反射率の値を、洗浄性能の尺度として測定する。デルタレミッション値を得るために、0mgの酵素タンパク質/Lを有する試験物を、ブランクとして使用する。好ましくは、機械的な動きを、例えば、布地を有する洗浄液の振盪、回転又は撹拌の形態で、洗浄工程中に適用する。
方法の項に記載されるAMSA試験方法によって、変異体及び対応する親アルファ−アミラーゼの洗浄性能を試験した。結果は、(変異体の性能−ブランクの性能)を(親の性能−ブランクの性能)で割り、100を乗じることによって得、ここでブランクは同じ条件下であるが、アルファ−アミラーゼの不在下の洗浄により得られた性能である。最後に、0.8〜1.2Mg/Lの2つの濃度の相対的性能の平均が算出された。
Tergo−To−Meter(TOM)は、12の異なる洗浄条件を同時に試験するように適用することができる中規模モデル洗浄システムである。TOMは、基本的に、それに沈められる12個の開放型金属ビーカーを備える、温度制御された大きな水層である。それぞれのビーカーは、小さなトップローダー式洗濯機1つを構成し、実験中、それぞれの洗濯機は、特定の洗剤/酵素系の溶液並びにその性能が試験される汚れた及び汚れていない布地を含有する。機械応力は、それぞれのビーカー内の液体を攪拌する攪拌アームを回転させることにより達成される。TOMビーカーは蓋がないため、TOM実験中にサンプルを取り出し、洗浄中に情報に関してオンラインでアッセイすることが可能である。
水の硬度は、CaCl2、MgCl2、及びNAHCO3を添加することにより、以下に記載される強度に調整された。洗浄溶液は、以下に記載するバケツの中で所望の洗剤量、温度、及び水硬度で調製された。10分間の磁気攪拌中に洗剤は溶解された。(洗浄溶液は調製後30〜60分以内に使用された)。
ヨーロッパ(EU)(実施例1A)及び北米(実施例1B)(US)液体洗剤におけるアルファ−アミラーゼの洗浄性能
試験された変異体及び対応する親アルファ−アミラーゼ(配列番号7)の洗浄性能は、上述のように試験された。結果は、(変異体の性能−ブランクの性能)を(親の性能−ブランクの性能)で割り、100を乗じることによって得、ここでブランクは、同じ条件下であるが、アルファ−アミラーゼの不在下の洗浄により得られた性能である。
3両親媒性アルコキシル化ポリマーは、−NHにつき24個のエトキシレート基、及び−NHにつき16個のプロポキシレート基を含有するように誘導化されるポリマーから調製されたポリエチレンイミン(MW 600)である。
液体洗浄中のアミラーゼ変異体の大規模な洗濯機性能評価
I.洗浄試験組成物の調製
この実験では、4つの試験組成物が液体洗剤処方2Aに基づき調製された。洗剤基材を、酵素を含有しない処方2Aから調製し、pH 8.2に仕上げた。
1Natalase(登録商標)は、Novozymes A/S,Bagsvaerd,Denmarkにより「Natalase 200L」として供給されるアミラーゼ酵素である。
2変異体3は、次の2つの欠失D183*+G184*を伴い、置換G109A、W140Y、N195F、V206Y、Y243F、E260G、及びG476Eを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476Eとも称される。
3変異体4は、次の2つの欠失D183*+G184*を伴い、置換T51I、G109A、W140Y、N195F、V206Y、Y243F、E260G、及びW439Rを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439Rとも称される。
4変異体5は、次の2つの欠失D183*+G184*を伴い、置換T51I、S52Q、N54K、G109A、W140Y、N195F、V206Y、Y243F、E260G、及びG476Eを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476Eとも称される。
5変異体6は、次の2つの欠失D183*+G184*及び置換W140Y、N195F、V206Y、Y243F、E260G、G304R、及びG476Kを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476Kとも称される。
6変異体7は、次の2つの欠失D183*+G184*、及び置換W140Y、N195F、V206Y、Y243F、E260G、W284R、及びG477Kを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477Kとも称される。
3つのアミラーゼ高感度の染み;5cm×5cmのPCS−28米デンプン、CS−128古米デンプン、及びCS−26コーンスターチ(Centre For Test materials,Netherlandsにより供給された)を、20cm×20cmの白い綿メリヤス(Warwick Equest,Durham,United Kingdomにより供給された)に付着させた。2つアミラーゼ高感度の染み:チリコンカーン及びハインツスパゲッティ(直径2.5cm)を、20cm×20cmの白い綿メリヤス(Warwick Equest,Durham,United Kingdomにより供給された)に付着させた。8つの複写物(4つの異なる機械に対して2つの複写物)をそれぞれの試験処方に使用した。
本方法は、Western Europe Hotpoint洗濯機、モデルAquarius WF541の使用を伴う。上述の試験処方は、上述の汚れ及びきれいなバラスト混合量を加えて、アミラーゼ高感度の染みを洗浄するために使用された。
液体洗浄中のアミラーゼ変異体の大規模な洗濯機性能評価
I.洗浄試験組成物の調製
この実験では、4つの試験組成物が、液体洗剤処方3Aに基づき調製された。洗剤基材を、酵素を含まない処方3Aから調製し、pH 8.2に仕上げた。
3両親媒性アルコキシル化ポリマーは、−NHにつき24個のエトキシレート基、及び−NHにつき16個のプロポキシレート基を含有するように誘導化されるポリマーから調製されたポリエチレンイミン(MW 600)である。
1Natalase(登録商標)は、Novozymes A/S,Bagsvaerd,Denmarkにより「Natalase 200L」として供給される。
2変異体5は、次の2つの欠失D183*+G184*を伴い、置換T51I、S52Q、N54K、G109A、W140Y、N195F、V206Y、Y243F、E260G、及びG476Eを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476Eとも称される。
3変異体6は、次の2つの欠失D183*+G184*を伴い、置換W140Y、N195F、V206Y、Y243F、E260G、G304R、及びG476Kを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476Kとも称される。
4変異体7は、次の2つの欠失D183*+G184*を伴い、置換W140Y、N195F、V206Y、Y243F、E260G、W284R、及びG477Kを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477Kとも称される。
5変異体8は、次の2つの欠失D183*+G184*を伴い、置換W140Y、N195F、V206Y、Y243F、E260G、W284F、及びG477Rを含む、野生型アミラーゼバチルス属722配列番号1の本発明のアミラーゼ変異体である。SP722+D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477Rとも称される。
3つのアミラーゼ高感度の染み;5cm×5cmのPCS−28米デンプン、PS−28米デンプン、及びCS−29タピオカデンプン(Centre For Test materials,Netherlandsにより供給される)を、20cm×20cmの白い綿メリヤス(Warwick Equest,Durham,United Kingdomにより供給された)に付着させた。1つアミラーゼ高感度の染み:ハインツスパゲッティ(直径2.5cm)を、20cm×20cmの白い綿メリヤス(Warwick Equest,Durham,United Kingdomにより供給された)に付着させた。1つアミラーゼ高感度の染み:グレービー(直径2.5cm)を、25cm×24cmの白い綿メリヤス(Accurate Product Development,Fairfield,Ohio,USAにより供給された)に付着させた。8つの複写物(4つの異なる機械に対して2つの複写物)をそれぞれの試験処方に使用した。
本方法は、North American Kenmore洗濯機モデル600シリーズの使用を伴う。上述の試験処方は、上述の汚れ及びきれいなバラスト混合量を加えて、アミラーゼ高感度の染みを洗浄するために使用された。
本発明は、ある場所、とりわけ表面又は布地をクリーニング及び/又は処理する方法を含む。一態様では、任意に洗浄する工程及び/又は表面若しくは布地をすすぐ工程、表面若しくは布地を本明細書に開示される任意の消費者製品と接触させて、次いで任意に表面若しくは布地を洗浄及び/又はすすぐ工程、を含む、そのような方法が開示される。
(実施例1〜6)
手洗い用又はトップローディング式洗濯機用に設計されている顆粒状洗濯洗剤組成物
フロントローディング式自動洗濯機用に設計されている顆粒状洗濯洗剤組成物
2−NHにつき20個のエトキシル基を有するポリエチレンイミン(MW=600)。
3両親媒性アルコキシル化グリース洗浄ポリマーは、1つの−NHにつき24個のエトキシレート基と、1つの−NHにつき16個のプロポキシレート基とを有するポリエチレンイミン(MW=600)である。
*本発明のアミラーゼは、洗剤100g当たりの活性酵素のmgとして示される。
**洗浄及び/又はトリートメント組成物の総重量に基づき、総量7%以下の水。
1ランダムグラフトコポリマーは、ポリエチレンオキシド主鎖と複数のポリビニルアセテート側鎖とを有する、ポリビニルアセテートグラフト化ポリエチレンオキシドコポリマーである。ポリエチレンオキシド主鎖の分子量は約6000であり、ポリ酢酸ビニルに対するポリエチレンオキシドの重量比は約40:60であり、エチレンオキシド単位50個当たりグラフト点は1個以下である。
2−NHあたり20個のエトキシル基を有するポリエチレンイミン(MW=600)。
3両親媒性アルコキシル化ポリマーは、−NHにつき24のエトキシレート基、及び−NHにつき16のプロポキシレート基を含有するように誘導化されるポリマーから調製されたポリエチレンイミン(MW 600)である。
平均脂肪族炭素鎖長を有するC11〜C18直鎖状アルキルベンゼンスルホン酸塩
C12〜18ジメチルヒドロキシエチル塩化アンモニウム
AE3Sは、C12〜15アルキルエトキシ(3)スルフェートである。
AE7は、7の平均エトキシル化度を有するC12〜15アルコールエトキシレートである。
AE9は、9の平均エトキシル化度を有するC12〜16アルコールエトキシレートである。
HSASは、米国特許第6,020,303号及び同第6,060,443号に開示される約16〜17の炭素鎖長の中分枝状一級アルキルサルフェートである。
ポリアクリレートMW 4500はBASFにより供給される。
カルボキシメチルセルロースは、CP Kelco,Arnhem,Netherlandsにより供給されるFinnfix(登録商標)Vである。
ホスホン酸塩キレート剤は、例えば、ジエチレンテトラアミン5酢酸(DTPA)ヒドロキシエタンジホスホン酸塩(HEDP)である。
Savinase(登録商標)、Natalase(登録商標)、Stainzyme(登録商標)、Lipex(登録商標)、Celluclean(商標)、Mannaway(登録商標)、及びWhitezyme(登録商標)は、すべてNovozymes,Bagsvaerd,Denmarkの製品である。
Purafect(登録商標)、Purafect Prime(登録商標)は、Genencor International,Palo Alto,California,USAの製品である。
蛍光増白剤1はTinopal(登録商標)AMSであり、蛍光増白剤2はTinopal(登録商標)CBS−Xであり、Direct Violet 9は、Pergasol(登録商標)であり、Violet BN−Z NOBSは、ノナノイルオキシベンゼンスルホン酸ナトリムである。
TAEDはテトラアセチルエチレンジアミンである。
S−ACMCは、C.I.リアクティブブルー19(商品名AZO−CM−CELLULOSE)と共役するカルボキシメチルセルロースである。
汚れ放出剤はRepel−o−tex(登録商標)PFである。
アクリル酸/マレイン酸コポリマーは分子量70,000であり、アクリル酸塩:マレイン酸塩比は70:30である。
EDDSはエチレンジアミン−N,N’−二コハク酸のナトリウム塩であり、(S,S)アイソマー抑泡剤疑集体はDow Corning,Midland,Michigan,USAにより供給される。
HSASは中分枝状アルキルサルフェートである。
リキティント(Liquitint)(登録商標)バイオレットCTは、ミリケン(Milliken)(米国サウスカロライナ州スパルタンバーグ)から供給されている。
以下に、本発明の複数区画単位用量の洗濯洗剤配合物が提供される。これらの実施例では、単位用量は3つの区画を有するが、同様の組成物を2、4、又は5区画で製造することもできる。区画を封入するために使用されるフィルムはポリビニルアルコールである。
Claims (18)
- 洗浄組成物であって、
(a)配列番号1の成熟ポリペプチドのG304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、及びG477からなる群から選択される位置に対応する2つ以上の位置での変性を含む、親アルファ−アミラーゼの変異体であって、それぞれの変性が、独立して、置換、欠失、又は挿入であり、前記変性の1つが、配列番号1の成熟ポリペプチドのE260の位置に対応する位置での置換であり、前記置換が、G、K、及びTからなる群から選択され、前記変異体が配列番号1の成熟ポリペプチドと少なくとも90%であるが、100%未満である配列同一性を有し、前記変異体がアルファ−アミラーゼ活性を有する、変異体と、
(b)0.01〜99.9重量%の量の洗浄補助剤と、
を含む、洗浄組成物。 - 前記変異体が、G304、W140、W189、D134、F262、W284、W347、W439、W469、G476、及びG477からなる群から選択される2つの位置での置換をさらに含む、請求項1に記載の組成物。
- 前記変異体が、G304、W140、W189、D134、F262、W284、W347、W439、W469、G476、及びG477からなる群から選択される3つの位置での置換をさらに含む、請求項1又は2に記載の組成物。
- 前記変異体が、G304、W140、及びG476からなる群から選択される1つ、2つ、又は3つの位置での置換をさらに含む、請求項1〜3のいずれか一項に記載の組成物。
- 前記変異体が、G304R、W140YF、W189EGT、D134E、W284DFR、W439RG、G476EK、G477EKMRからなる群から選択される1つ以上の置換をさらに含む、請求項1〜4のいずれか一項に記載の組成物。
- 前記変異体が、G304R、W140YF、及びG476EKからなる群から選択される1つ、2つ、又は3つの位置での置換をさらに含む、請求項1〜5のいずれか一項に記載の組成物。
- 前記変異体が、E260Gの位置での置換を含み、かつG304R、W140Y、及びG476Kからなる群から選択される位置での1つ、2つ、又は3つの置換を含む、請求項6に記載の組成物。
- 前記変異体が、N195F、V206Y、Y243F、G109A、G273DV、G337N、K72R、R181H、S303G、及びY100Iからなる群から選択される1つ以上の置換を更に含む、請求項1〜7のいずれか一項に記載の組成物。
- 前記変異体の前記変性の数が、2〜12である、請求項1〜8のいずれか一項に記載の組成物。
- 前記変異体が、
W140Y+N195F+V206Y+Y243F+E260G+G477E、
W140Y+N195F+V206Y+Y243F+E260T+W284D、
G109A+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G、
N195F+V206Y+Y243F+E260K+W284D、
W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+W189G+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+S303G、
W140Y+W189T+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+W284D、
Y100I+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+G337N、
W140Y+N195F+V206Y+Y243F+E260G+W439R、
G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
N195F+V206Y+Y243F+E260G+W284D
からなる群から選択される、前記配列番号1のポリペプチドの位置に対応する位置に置換を含む、請求項1〜9のいずれか一項に記載の組成物。 - 前記変異体が、
W140Y+N195F+V206Y+Y243F+E260G+G477E、
W140Y+N195F+V206Y+Y243F+E260T+W284D、
G109A+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G、
N195F+V206Y+Y243F+E260K+W284D、
W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+W189G+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+S303G、
W140Y+W189T+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+W284D、
Y100I+W140Y+N195F+V206Y+Y243F+E260G、
W140Y+N195F+V206Y+Y243F+E260G+G337N、
W140Y+N195F+V206Y+Y243F+E260G+W439R、
G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
N195F+V206Y+Y243F+E260G+W284D
からなる群から選択される、前記配列番号1のポリペプチドの位置に対応する位置の置換からなる、請求項1〜10のいずれか一項に記載の組成物。 - 前記変異体が、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G477E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260T+W284D、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+N195F+V206Y+Y243F+E260K+W284D、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+W189G+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+S303G、
D183*+G184*+W140Y+W189T+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284D、
D183*+G184*+Y100I+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G337N、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
D183*+G184*+N195F+V206Y+Y243F+E260G+W284D
からなる群から選択される前記配列番号1のポリペプチドの位置に対応する位置での変性を含む、請求項1〜11のいずれか一項に記載の組成物。 - 前記変異体が、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G477E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260T+W284D、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+N195F+V206Y+Y243F+E260K+W284D、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+W189G+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+S303G、
D183*+G184*+W140Y+W189T+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284D、
D183*+G184*+Y100I+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G337N、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G、
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G、
D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R、
D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K、
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R、及び
D183*+G184*+N195F+V206Y+Y243F+E260G+W284D
からなる群から選択される、前記配列番号1のポリペプチドの位置に対応する位置での変性からなる、請求項1〜12のいずれか一項に記載の組成物。 - 前記変異体が、配列番号1の位置G182*+D183*又はD183*+G184*に対応する位置に欠失を更に含む、請求項1〜13のいずれか一項に記載の組成物。
- 前記変異体が、配列番号1の位置N195F+V206Y+Y243Fに対応する位置での置換を更に含む、請求項1〜14のいずれか一項に記載の組成物。
- 前記洗浄補助剤が、(i)香料マイクロカプセル、(ii)布地色相剤、(iii)プロテアーゼ、(iv)両親媒性洗浄用ポリマー、(v)リパーゼ、又は(vi)それらの混合物からなる群から選択される1つ以上を含む、請求項1〜15のいずれか一項に記載の組成物。
- 前記組成物が、液体洗濯洗剤組成物又は単位用量洗剤組成物である、請求項1〜16のいずれか一項に記載の組成物。
- 表面を処理する方法であって、
(i)水及び請求項1〜17のいずれか一項に記載の組成物を含む水性洗浄溶液を形成する工程と、
(ii)40℃以下の温度の水性洗浄溶液で、前記表面を処理する工程と、
(iii)前記表面をすすぐ工程と、
を含む、方法。
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US20160177238A1 (en) | 2016-06-23 |
FI3121270T3 (fi) | 2023-11-22 |
CN112680432A (zh) | 2021-04-20 |
EP2540825B1 (en) | 2016-10-12 |
CN103703126B (zh) | 2021-07-27 |
MX346999B (es) | 2017-04-06 |
DK3121270T3 (da) | 2023-11-20 |
MX2013015178A (es) | 2014-03-31 |
ZA201309188B (en) | 2015-09-30 |
EP3121270A3 (en) | 2017-03-08 |
US9284544B2 (en) | 2016-03-15 |
HUE032374T2 (en) | 2017-09-28 |
DK2540825T3 (en) | 2017-01-16 |
ES2608964T3 (es) | 2017-04-17 |
EP2540824A1 (en) | 2013-01-02 |
CN103703126A (zh) | 2014-04-02 |
PL3121270T3 (pl) | 2024-02-12 |
EP2540825A3 (en) | 2013-03-20 |
ES2966391T3 (es) | 2024-04-22 |
PL2540825T3 (pl) | 2017-04-28 |
RU2617954C2 (ru) | 2017-04-28 |
EP3121270A2 (en) | 2017-01-25 |
AR086831A1 (es) | 2014-01-22 |
BR112013033581A2 (pt) | 2017-10-03 |
WO2013003659A1 (en) | 2013-01-03 |
US20130000055A1 (en) | 2013-01-03 |
RU2013156968A (ru) | 2015-08-10 |
JP2014523246A (ja) | 2014-09-11 |
EP2540825A2 (en) | 2013-01-02 |
EP3121270B1 (en) | 2023-11-01 |
US9670436B2 (en) | 2017-06-06 |
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