Title:
化学的に修飾された突然変異体酵素およびその産生方法とスクリーニング方法
Document Type and Number:
Japanese Patent JP4475371
Kind Code:
B2
Abstract:
The present invention relates to a method for screening chemically modified mutant enzymes for amidase and/or esterase activity. This method includes providing a chemically modified mutant enzyme with one or more amino acid residues from an enzyme being replaced by cysteine residues, where at least some of the cysteine residues are modified by replacing thiol hydrogen in the cysteine residues with a thiol side chain, contacting the chemically modified mutant enzyme with a substrate for an amidase and/or a substrate for an esterase, and determining whether the chemically modified mutant enzyme exhibits amidase and/or esterase activity. The present invention also relates to chemically modified mutant enzymes and a method of producing them where one or more amino acid residues from an enzyme are replaced by cysteine residues, and the cysteine residues are modified by replacing at least some of the thiol hydrogen in the cysteine residue with a thiol side chain to form the chemically modified mutant enzyme. The thiol side chain is selected from the group consisting of -SCH2(p-CH3-C6H4), -SCH2(p-OCH3-C6H4), -SCH2(p-CF3-C6H4), and -SCH2(2,4-diNO2-C6H3).
Inventors:
Jones, Jay Brian
Plettner, Erica
Plettner, Erica
Application Number:
JP2000528303A
Publication Date:
June 09, 2010
Filing Date:
January 21, 1999
Export Citation:
Assignee:
Genencor International Inc.
International Classes:
C12N9/14; C12N9/16; C11D3/00; C11D3/386; C12N9/48; C12N9/50; C12N9/54; C12N9/56; C12N9/80; C12N15/09; C12P21/06; C12Q1/34; C12Q1/44; D06M16/00
Domestic Patent References:
| JP5507402A | ||||
| JP58144105A | ||||
| JP63502639A |
Foreign References:
| WO1996005739A1 |
Other References:
BERGLUND,JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,1997年 6月 4日,V119N22,P5265-5266
BERGLUND,BIOORGANIC AND MEDICINAL CHEMISTRY LETTERS,1996年11月 1日,V6N21,P2507-2512
J. Am. Chem. Soc.,1991年,Vol.113,p.1026-1030
J. Mol. Biol.,1992年,Vol.223,p.427-445
Biochemistry,1991年,Vol.30,p.6142-6148
BERGLUND,BIOORGANIC AND MEDICINAL CHEMISTRY LETTERS,1996年11月 1日,V6N21,P2507-2512
J. Am. Chem. Soc.,1991年,Vol.113,p.1026-1030
J. Mol. Biol.,1992年,Vol.223,p.427-445
Biochemistry,1991年,Vol.30,p.6142-6148
Attorney, Agent or Firm:
Yanagita Seiji
Go Sakuma
Go Sakuma