1. Purification and properties of two malate dehydrogenases from Candida sp. N-16 grown on methanol
- Author
-
Jun Yoshikawa, Takaaki Fujii, Hirofumi Shinoyama, and Katsura Seki
- Subjects
Oxaloacetic Acid ,Applied Microbiology and Biotechnology ,Biochemistry ,Isozyme ,Malate dehydrogenase ,Analytical Chemistry ,Malate Dehydrogenase ,Citrate synthase ,Protein Structure, Quaternary ,Molecular Biology ,Candida ,chemistry.chemical_classification ,biology ,Methanol ,Organic Chemistry ,General Medicine ,Metabolism ,NAD ,Isoenzymes ,Molecular Weight ,Kinetics ,Oxaloacetate decarboxylase ,Enzyme ,chemistry ,biology.protein ,NAD+ kinase ,Biotechnology ,Homotetramer - Abstract
Two malate dehydrogenases (MDH-M1 and MDH-M2) were found in a methanol-using yeast, Candida sp. N-16. MDH-M2 was induced with methanol. These enzymes were purified as electrophoretically and isoelectrophoretically homogeneous proteins. The molecular weights of MDH-M1 and MDH-M2 were estimated to be about 78,000 (homodimer) and 160,000 (homotetramer). Several kinetic properties were significantly different between the two enzymes. The value (2.07) of Vmax(oxaloacetate)/Vmax(malate) and Kcats (555 s(-1) for oxaloacetate, 481 s(-1) for NADH) of MDH-M2 were higher than the ratio (1.37) of Vmax and Kcats (241 s(-1) for oxaloacetate, 271 s(-1) for NADH) of MDH-M1, respectively. The activity of MDH-M2 was inhibited by a high concentration of NAD+ and the activity of MDH-M1 by oxaloacetate.
- Published
- 2001