JP2009543577A - 幅広い温度におけるプロテアーゼ変異体の活性 - Google Patents
幅広い温度におけるプロテアーゼ変異体の活性 Download PDFInfo
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- JP2009543577A JP2009543577A JP2009520754A JP2009520754A JP2009543577A JP 2009543577 A JP2009543577 A JP 2009543577A JP 2009520754 A JP2009520754 A JP 2009520754A JP 2009520754 A JP2009520754 A JP 2009520754A JP 2009543577 A JP2009543577 A JP 2009543577A
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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)
- C12N9/48—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
-
- 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
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- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
Abstract
【選択図】
図1A
Description
H2N−
脂肪族
天然
非極性(G、A、P、L、I、V)
極性(C、M、S、T、N、Q)
電荷
負電荷(D、E)
正電荷(K、R)
芳香族
(F、H、W、Y)
任意のアミノ酸は同じカテゴリー、特に同じラインの他のアミノ酸で置換される。更に、極性アミノ酸、N、Qは電荷アミノ酸に置換するか、又はこれらにより置換される。本発明の目的のために、プロテアーゼのアニオン性質を高めることとなる、特に活性部位に直接含まれない置換が特に好ましい。
G116 I、V、L
S126を任意のアミノ酸、P127を任意のアミノ酸、S128を任意のアミノ酸、
S160を芳香族又は中性脂肪族アミノ酸、
A166を電荷のある、特定のアミノ酸
M169を天然の脂肪族、好ましくは非極性のアミノ酸
N212 アニオンアミノ酸
M216脂肪族、極性、特に、S、T、N、Q
実施例1
PB92プロテアーゼ変異体の構築
A.変異誘発ベクター13M1の構築、
B.変異手順
C.pM58Ecoの構築及びベクター内での変異DNAフラグメントのサブクローニング
A. 変異誘発ベクター13M1の構築
変異誘発手順
C.pM58Ecoの構築及びベクター内での変異DNAフラグメントのサブクローニング
D.PB92変異体の生成
実施例2
バチルスクラウシ(B.clausii)における変異体の発現
PB92プロテアーゼ陰性融合体:PBT125の形質転換手順
発酵条件及びプロテアーゼ生成
実施例3
精製されたプロテアーゼの精製度を確認するための解析技術
実施例4
プロテアーゼ濃度の決定
実施例5
野生型変異プロテアーゼの動的パラメーターの決定
実施例6
洗浄性能試験
ステンレススチール上の黄身の汚れ
これらの実験について、欧州規格(EN)50242、フォーム1495、No.0219、直径19cmデザートプレート(Arzberg, white, glazed porcelain)を用いた。トータル225gの赤みのブタ及びビーフ(当量の混合物)を目視可能な脂肪を取り除き、細かく刻み、冷却した。この混合物を2回、ひき肉器にかけた。35℃以上の温度を避けた。その後、この225gのひき肉を75gのタマゴと混ぜた(黄身と白身の混合物)。調製後、用いる前に3ヶ月間―18℃で保存した。これらは互換的に使用可能であるので、ブタ肉が使用できない場合、牛肉を用いた。
ステンレススチール上のタマゴ/ミルクの汚れ
洗浄装置及び条件
行う事ができることは、当業者に理解されており、そのような変更は本発明の範囲内である。
Claims (12)
- 修飾されたスブチリシンを含む、食器洗浄用組成物であって、
前記スブチリシンが配列番号2で定義する配列において、少なくとも1つの置換を含み、各置換位置はスブチリシンBPN’のアミノ酸配列の位置に対応し、これらの置換がG118、S128、P129、及びS166の位置から選択される、組成物。 - 前記修飾されたスブチリシンがG118、S128、P129、及びS130の位置における置換を含む請求項1の組成物。
- 前記修飾されたスブチリシンがG118Vと他の1つの追加的な変異を含む請求項1の食器洗浄用組成物。
- 前記追加的な変異が、S128F、S128L、S128N、S128R、S128V、P129E、P129L、P129M、P129N、P129L、P129Q、P129S、S130A、S130K、S130P、S130T、S130V、及びS166Dからなる群より選択される請求項3の食器洗浄用組成物。
- 前記修飾されたスブチリシンがS128、P129、及びS130の置換位置を含む、請求項1の食器洗浄用組成物。
- 前記置換がS128C、S128R、P129Q、P129R,S130D、及びS130Gから選択される、請求項5の食器洗浄用組成物。
- 前記修飾されたスブチリシンのアミノ酸配列が配列番号3で定義される、請求項1の食器洗浄用組成物。
- 修飾されたスブチリシンを含む、食器洗浄用組成物であって、
前記スブチリシンが配列番号2で定義する配列において、1つの置換を含み、各置換位置はスブチリシンBPN’のアミノ酸配列の位置に対応し、前記置換がS130Tである、食器洗浄用組成物。 - 請求項1で定義される修飾されたスブチリシンをコードする単離アミノ酸。
- 請求項9の単離された核酸を含むベクター。
- 請求項10のベクターを含む宿主細胞。
- 請求項1で定義される少なくとも1つの修飾されたスブチリシンと食器とを提供する工程、並びに前記食器を洗浄に効果的な条件下で、前記食器を前記修飾されたスブチリシンに接触せる工程とを含む、食器洗浄方法。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US83173206P | 2006-07-18 | 2006-07-18 | |
PCT/US2007/015642 WO2008010925A2 (en) | 2006-07-18 | 2007-07-09 | Protease variants active over a broad temperature range |
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JP2015193074A Division JP2016052308A (ja) | 2006-07-18 | 2015-09-30 | 幅広い温度におけるプロテアーゼ変異体の活性 |
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JP2009543577A true JP2009543577A (ja) | 2009-12-10 |
JP2009543577A5 JP2009543577A5 (ja) | 2014-02-27 |
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JP2009520754A Pending JP2009543577A (ja) | 2006-07-18 | 2007-07-09 | 幅広い温度におけるプロテアーゼ変異体の活性 |
JP2015193074A Pending JP2016052308A (ja) | 2006-07-18 | 2015-09-30 | 幅広い温度におけるプロテアーゼ変異体の活性 |
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Country Status (13)
Country | Link |
---|---|
US (6) | US20080090747A1 (ja) |
EP (1) | EP2059591B1 (ja) |
JP (2) | JP2009543577A (ja) |
KR (2) | KR20150115023A (ja) |
CN (2) | CN101511998A (ja) |
AU (1) | AU2007275864B2 (ja) |
BR (1) | BRPI0714329B1 (ja) |
CA (1) | CA2658396C (ja) |
DK (1) | DK2059591T3 (ja) |
MX (1) | MX2009000537A (ja) |
NZ (1) | NZ573467A (ja) |
RU (1) | RU2509152C2 (ja) |
WO (1) | WO2008010925A2 (ja) |
Cited By (4)
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JP2010507001A (ja) * | 2006-10-16 | 2010-03-04 | ダニスコ・ユーエス・インク、ジェネンコー・ディビジョン | 非リン酸系食器洗浄剤 |
JP2016052308A (ja) * | 2006-07-18 | 2016-04-14 | ダニスコ・ユーエス・インク、ジェネンコー・ディビジョン | 幅広い温度におけるプロテアーゼ変異体の活性 |
JP2021507019A (ja) * | 2017-12-19 | 2021-02-22 | ザ プロクター アンド ギャンブル カンパニーThe Procter & Gamble Company | 自動食器洗浄洗剤組成物 |
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JP2003516751A (ja) * | 1999-12-15 | 2003-05-20 | ノボザイムス アクティーゼルスカブ | 卵の染みに及ぼす洗浄性能改良を示すズブチラーゼ変異体 |
JP2006505285A (ja) * | 2002-11-06 | 2006-02-16 | ノボザイムス アクティーゼルスカブ | サブチラーゼ変異体 |
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JP2010507001A (ja) * | 2006-10-16 | 2010-03-04 | ダニスコ・ユーエス・インク、ジェネンコー・ディビジョン | 非リン酸系食器洗浄剤 |
JP2021507019A (ja) * | 2017-12-19 | 2021-02-22 | ザ プロクター アンド ギャンブル カンパニーThe Procter & Gamble Company | 自動食器洗浄洗剤組成物 |
JP7196178B2 (ja) | 2017-12-19 | 2022-12-26 | ザ プロクター アンド ギャンブル カンパニー | 自動食器洗浄洗剤組成物 |
US11834634B2 (en) | 2017-12-19 | 2023-12-05 | The Procter & Gamble Company | Phosphate-free automatic dishwashing detergent compositions having a protease and a complexing agent |
Also Published As
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BRPI0714329A2 (pt) | 2013-03-05 |
CN104232365A (zh) | 2014-12-24 |
US20130323816A1 (en) | 2013-12-05 |
RU2509152C2 (ru) | 2014-03-10 |
CN101511998A (zh) | 2009-08-19 |
AU2007275864B2 (en) | 2013-05-02 |
US20140302590A1 (en) | 2014-10-09 |
EP2059591B1 (en) | 2012-09-05 |
CA2658396C (en) | 2019-07-16 |
WO2008010925A2 (en) | 2008-01-24 |
KR20090031906A (ko) | 2009-03-30 |
DK2059591T3 (da) | 2012-10-29 |
EP2059591A2 (en) | 2009-05-20 |
US20080090747A1 (en) | 2008-04-17 |
MX2009000537A (es) | 2009-01-28 |
AU2007275864A1 (en) | 2008-01-24 |
CA2658396A1 (en) | 2008-01-24 |
US20190078070A1 (en) | 2019-03-14 |
US20190338267A1 (en) | 2019-11-07 |
US20110059517A1 (en) | 2011-03-10 |
JP2016052308A (ja) | 2016-04-14 |
RU2009105503A (ru) | 2010-08-27 |
KR20150115023A (ko) | 2015-10-13 |
WO2008010925A3 (en) | 2008-04-10 |
NZ573467A (en) | 2012-02-24 |
BRPI0714329B1 (pt) | 2017-09-12 |
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