Purification and properties of oxaloacetate decarboxylase from Corynebacterium glutamicum.

TitlePurification and properties of oxaloacetate decarboxylase from Corynebacterium glutamicum.
Publication TypeJournal Article
Year of Publication1995
AuthorsJetten, MS, Sinskey, AJ
JournalAntonie Van Leeuwenhoek
Date Published1995
KeywordsCarboxy-Lyases, Cations, Corynebacterium, Electrophoresis, Polyacrylamide Gel, Kinetics, Manganese, Molecular Weight, Pyruvate Kinase, Substrate Specificity

Oxaloacetate (OAA) decarboxylase (E.C. was isolated from Corynebacterium glutamicum. In five steps the enzyme was purified 300-fold to apparent homogeneity. The molecular mass estimated by gel filtration was 118 +/- 6 kDa. SDS-PAGE showed a single subunit of 31.7 KDa, indicating an alpha 4 subunit structure for the native enzyme. The enzyme catalyzed the decarboxylation of OAA to pyruvate and CO2, but no other alpha-ketoacids were used as substrate. The cation Mn2+ was required for full activity, but could be substituted by Mg2+, CO2+, Ni2+ and Ca2+. Monovalent ions like Na+, K+ or NH4+ were not required for activity. The enzyme was inhibited by Cu2+, Zn2+, ADP, coenzyme A and succinate. Avidin did not inhibit the enzyme activity, indicating that biotin is not involved in decarboxylation of OAA. Analysis of the kinetic properties revealed a Km for OAA of 2.1 mM and a Km of 1.2 mM for Mn2+. The Vmax was 158 mumol of OAA converted per min per mg of protein, which corresponds to an apparent kcat of 311 s-1.

Alternate JournalAntonie Van Leeuwenhoek
Citation Key193
PubMed ID7771770