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Research article

Article

Research article

J. Microbiol. Biotechnol. 2012; 22(1): 141-146

Published online January 28, 2012 https://doi.org/10.4014/jmb.1107.07058

Copyright © The Korean Society for Microbiology and Biotechnology.

Functional Expression and Characterization of Recombinant NADPH-P450 Reductase from Malassezia globosa

Hwayoun Lee 1, Hyoung-Goo Park 1, Young-Ran Lim 1, Im-Soon Lee 1, Beom Joon Kim 2, Cheul-Hun Seong 3, Young-Jin Chun 3 and Donghak Kim 1*

1Department of Biological Sciences and Center for Biotechnology Research at UBITA (CBRU), Konkuk University, Seoul, 143-701, Korea, 2Department of Dermatology, College of Medicine, Chung-Ang University, Seoul, 156-755, Korea, 3College of Pharmacy, Chung-Ang University, Seoul, 156-756, Korea.

Received: July 26, 2011; Accepted: September 17, 2011

Abstract

Malassezia globosa is a common pathogenic fungus that
causes skin diseases including dandruff and seborrheic
dermatitis in humans. Analysis of its genome identified a
gene (MGL_1677) coding for a putative NADPH-P450
reductase (NPR) to support the fungal cytochrome P450
enzymes. The heterologously expressed recombinant M.
globosa NPR protein was purified, and its functional
features were characterized. The purified protein generated
a single band on SDS-PAGE at 80.74 kDa and had an
absorption maximum at 452 nm, indicating its possible
function as an oxidized flavin cofactor. It evidenced
NADPH-dependent reducing activity for cytochrome c or
nitroblue tetrazolium. Human P450 1A2 and 2A6 were
able to successfully catalyze the O-deethylation of 7-
ethoxyresorufin and the 7-hydroxylation of coumarin,
respectively, with the support of the purified NPR. These
results demonstrate that purified NPR is an orthologous
reductase protein that supports cytochrome P450 enzymes
in M. globosa.

Keywords: Reductase, Cytochrome P450, Malassezia globosa, NADPH, 7-Ethoxyresorufin, Coumarin