Fluorogenic assay for penicillin G acylase activity

被引：11

作者：

Ninkovic, M ^{[1
]}

Riester, D ^{[1
]}

Wirsching, F ^{[1
]}

Dietrich, R ^{[1
]}

Schwienhorst, A ^{[1
]}

机构：

[1] Inst Mikrobiol & Genet, Abt Mol Genet & Praeparat Mol Biol, D-37077 Gottingen, Germany

来源：

ANALYTICAL BIOCHEMISTRY | 2001年 / 292卷 / 02期

关键词：

fluorogenic substrates; high-throughput screening assay; penicillin G acylase;

D O I：

10.1006/abio.2001.5078

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple, highly sensitive, and rapid assay for high-throughput screening of penicillin G acylase-producing bacteria is presented. The method is based on the specific release of fluorescent 7-amino-4-methyl-coumarin through cleavage of phenylacetyl-4-methyl-coumaryl-7-amide by penicillin G acylase. The present method is suitable for screening pure enzymes as well as various penicillin G acylases like those from Escherichia coli, Proteus rettgeri, and Kluyvera citrophila in cell extracts. In addition, the new substrate was used for rapid assay of amidase activity in nondenaturing polyacrylamide gels. (C) 2001 Academic Press.

引用

页码：228 / 233

页数：6

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