A spectrophotometric assay for measuring and detecting an epoxide hydrolase activity.
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.
Mateo
Cesar
C
Groupe Biocatalyse et Chimie Fine, UMR CNRS 6111, Faculté des Sciences de Luminy, Université de la Méditerranée, Case 901, 163 Avenue de Luminy, 13288 Marseille cedex 9, France.
Archelas
Alain
A
Furstoss
Roland
R
D001234:Aspergillus niger / Q000201:enzymology; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000378:metabolism*; D005026:Ethylene Glycols; D007700:Kinetics; D010084:Oxidation-Reduction; D010504:Periodic Acid; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
D005026:Ethylene Glycols; D010504:Periodic Acid; C031474:styrene glycol; C009288:metaperiodate; D004851:Epoxide Hydrolases