Electron transfer ability from NADH to menaquinone and from NADPH to oxygen of type II NADH dehydrogenase of Corynebacterium glutamicum.
Type II NADH dehydrogenase of Corynebacterium glutamicum (NDH-2) was purified from an ndh overexpressing strain. Purification conferred 6-fold higher specific activity of NADH:ubiquinone-1 oxidoreductase with a 3.5-fold higher recovery than that previously reported (K. Matsushita et al., 2000). UV-visible and fluorescence analyses of the purified enzyme showed that NDH-2 of C. glutamicum contained non-covalently bound FAD but not covalently bound FMN. This enzyme had an ability to catalyze electron transfer from NADH and NADPH to oxygen as well as various artificial quinone analogs at neutral and acidic pHs respectively. The reduction of native quinone of C. glutamicum, menaquinone-2, with this enzyme was observed only with NADH, whereas electron transfer to oxygen was observed more intensively with NADPH. This study provides evidence that C. glutamicum NDH-2 is a source of the reactive oxygen species, superoxide and hydrogen peroxide, concomitant with NADH and NADPH oxidation, but especially with NADPH oxidation. Together with this unique character of NADPH oxidation, phylogenetic analysis of NDH-2 from various organisms suggests that NDH-2 of C. glutamicum is more closely related to yeast or fungal enzymes than to other prokaryotic enzymes.
Nantapong
Nawarat
N
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan.
Otofuji
Asuka
A
Migita
Catharina T
CT
Adachi
Osao
O
Toyama
Hirohide
H
Matsushita
Kazunobu
K
D000968:Antimycin A / Q000494:pharmacology; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics; D004579:Electron Transport; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009243:NAD / Q000378:metabolism*; D009245:NADH Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010802:Phylogeny; D013481:Superoxides / Q000378:metabolism; D024482:Vitamin K 2 / Q000378:metabolism*
D009243:NAD; D024482:Vitamin K 2; D013481:Superoxides; D009249:NADP; D000968:Antimycin A; D006861:Hydrogen Peroxide; D009245:NADH Dehydrogenase; D010100:Oxygen