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Fluorogenic Assay for Penicillin G Acylase Activity

✍ Scribed by Milena Ninkovic; Daniel Riester; Frank Wirsching; Rüdiger Dietrich; Andreas Schwienhorst

Publisher
Elsevier Science
Year
2001
Tongue
English
Weight
85 KB
Volume
292
Category
Article
ISSN
0003-2697

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✦ Synopsis

A simple, highly sensitive, and rapid assay for highthroughput 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.

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