✦ LIBER ✦

An Assay of Flavin-Containing Monooxygenase Activity with Benzydamine N-Oxidation

✍ Scribed by A. Kawaji; K. Ohara; E. Takabatake

Publisher: Elsevier Science
Year: 1993
Tongue: English
Weight: 323 KB
Volume: 214
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.1993.1515

No coin nor oath required. For personal study only.

✦ Synopsis

An assay system of flavin-containing monooxygenase was developed by fluorometric determination of benzydamine (BZY) N-oxidation with HPLC. The apparent Km value for the formation of BZY N-oxide from BZY by rat liver microsomes was similar to that by purified FMO. The Km and Vmax values for the formation of N-desmethylbenzydamine (Nor-BZY) by rat liver microsomes were about 50 times greater and 2000 times less, respectively, than those of BZY N-oxide. Nor-BZY was not formed upon incubation with purified enzyme. BZY N-oxidation activity was completely inhibited both in the absence of NADPH and by heat inactivation. The reaction was inhibited in the presence of 0.5 mM thiourea, but 2 mM SKF-525A did not affect BZY N-oxidation. Moreover, rabbit antibody raised against the rat enzyme inhibited BZY N-oxidation. These results are in accord with a simple, rapid, and sensitive assay for the enzyme.

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