Purification and characterization of malate dehydrogenase from Corynebacterium glutamicum.
The malate dehydrogenase (MDH) (EC 1.1.1.37) from Corynebacterium glutamicum (Brevibacterium flavum) ATCC14067 was purified to homogeneity. Its amino-terminal sequence (residues 1 to 8) matched the sequence (residues 2 to 9) of the MDH from C. glutamicum (GenBank accession no. CAC83073). The molecular mass of the native enzyme was 130 kDa. The protein was a homotetramer, with a 33-kDa subunit molecular mass. The enzyme was almost equally active both for NADU and NADPH as coenzyme on the bases of the k(cat) values at pH 6.5 which is the optimum pH for the both coenzymes. Plotting of the logarithms of the 1/Km, k(cat), and k(cat)/K(m) values with respect to oxalacetate against pH lead to speculation that imidazolium is possibly a functional group in the active center of the enzyme. Citrate activated the enzyme in the oxidation of malate to oxalacetate and inhibited it in the reverse reaction.
Genda
Tomoko
T
Biological Institute, Faculty ofEducation, Yamaguchi University, 1677-1 Yoshida, Yamaguchi-shi 753-8513, Japan.
Nakamatsu
Tsuyoshi
T
Ozak
Hachiro
H