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A spectrophotometric assay for measuring and detecting an epoxide hydrolase activity.

Analytical biochemistry (2003-03-14)
Cesar Mateo, Alain Archelas, Roland Furstoss
ABSTRACT

In this paper we report the development of a novel and simple spectrophotometric assay which allows one to achieve the continuous, rapid, sensitive, and accurate determination of an epoxide hydrolase activity. This assay is based on the elaboration of a coupled enzymatic/chemical methodology which allows quantification of the enzymatic activity within 3min, and offers good sensitivity of about 10 micro Mmin(-1). Applicability of this test to some other aromatic epoxides has been shown and some limitations have also been explored. This assay should be particularly useful for different applications, for example (a) activity localization during purification of such enzymes, (b) very rapid determination of kinetic constants, and (c) high-throughput screening experiments.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(R)-(−)-1-Phenyl-1,2-ethanediol, 99%
Sigma-Aldrich
1-Phenyl-1,2-ethanediol, 97%
Sigma-Aldrich
(S)-(+)-1-Phenyl-1,2-ethanediol, 99%