Kinetic isotope effect of the L-phenylalanine oxidase from Pseudomonas sp. P-501.

Abstract:

:Pseudomonas L-phenylalanine oxidase (deaminating and decarboxylating) mainly catalyzes oxygenation when L-phenylalanine is used as the substrate, but oxidation when L-methionine is used as the substrate. Using [C(alpha)-H]-DL-methionine and [C(alpha)-D]-DL-methionine as substrate, the reductive half reaction of FAD cofactor of enzyme has been studied by stopped-flow spectrophotometry. The rate of reduction of FAD cofactor has a kinetic isotope effect (KIE) of 5.4 and 4.1 in the absence and presence of 30% glycerol, respectively. The KIE is independent of temperature, but the rates of the reductive half reaction are dependent on temperature, indicating that thermally induced motion at the active site drives the H-transfer reaction by H-tunneling.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Ohta Y,Mukouyama EB,Suzuki H

doi

10.1093/jb/mvj049

keywords:

subject

Has Abstract

pub_date

2006-03-01 00:00:00

pages

551-5

issue

3

eissn

0021-924X

issn

1756-2651

pii

139/3/551

journal_volume

139

pub_type

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