Purification and properties of NADH oxidase from Bacillus megaterium.

Abstract:

:NADH oxidase, which catalyzes the oxidation of NADH, with the consumption of a stoichiometric amount of oxygen, to NAD+ and hydrogen peroxide was purified from Bacillus megaterium by 5'-AMP Sepharose affinity chromatography to homogeneity. The enzyme is a dimeric protein containing 1 mol of FAD per mol of subunit, Mr = 52,000. The absorption maxima of the native enzyme (oxidized form) were found at 270, 383, and 450 with a shoulder at 475 nm in 50 mM KPi buffer, pH 7.0. The visible absorption bands at 383 and 450 nm disappeared on the addition of NADH under anaerobic conditions and reappeared upon the introduction of air. Thus, the non-covalently bound FAD functioned as a prosthetic group for the enzyme. We tentatively named this new enzyme NADH oxidase (NADH:oxygen oxidoreductase, hydrogen peroxide forming). This enzyme stereospecifically oxidizes the pro-S hydrogen at C-4 of the pyridine ring of NADH.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Saeki Y,Nozaki M,Matsumoto K

doi

10.1093/oxfordjournals.jbchem.a135411

subject

Has Abstract

pub_date

1985-12-01 00:00:00

pages

1433-40

issue

6

eissn

0021-924X

issn

1756-2651

journal_volume

98

pub_type

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