NADH dehydrogenase of Corynebacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH.
NADPH oxidase activity, in addition to NADH oxidase activity, has been shown to be present in the respiratory chain of Corynebacterium glutamicum. In this study, we tried to purify NADPH oxidase and NADH dehydrogenase activities from the membranes of C. glutamicum. Both the enzyme activities were simultaneously purified in the same fraction, and the purified enzyme was shown to be a single polypeptide of 55 kDa. The N-terminal sequence of the enzyme was consistent with the sequence deduced from the NADH dehydrogenase gene of C. glutamicum, which has been sequenced and shown to be a homolog of NADH dehydrogenase II. In addition to high NADH-ubiquinone-1 oxidoreductase activity at neutral pH, the purified enzyme showed relatively high NADPH oxidase and NADPH-ubiquinone-1 oxidoreductase activities at acidic pH. Thus, NADH dehydrogenase of C. glutamicum was shown to be rather unique in having a relatively high reactivity toward NADPH.
Matsushita
K
K
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Yamaguchi, Japan. kazunobu@agr.yamaguchi-u.ac.jp
Otofuji
A
A
Iwahashi
M
M
Toyama
H
H
Adachi
O
O
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000201:enzymology; D003352:Corynebacterium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D010084:Oxidation-Reduction; D017422:Sequence Analysis, DNA
D009243:NAD; D009249:NADP; D019255:NADPH Oxidases; D009245:NADH Dehydrogenase