Title | Purification and properties of oxaloacetate decarboxylase from Corynebacterium glutamicum. |
Publication Type | Journal Article |
Year of Publication | 1995 |
Authors | Jetten, MS, Sinskey, AJ |
Journal | Antonie Van Leeuwenhoek |
Volume | 67 |
Issue | 2 |
Pagination | 221-7 |
Date Published | 1995 |
ISSN | 0003-6072 |
Keywords | Carboxy-Lyases, Cations, Corynebacterium, Electrophoresis, Polyacrylamide Gel, Kinetics, Manganese, Molecular Weight, Pyruvate Kinase, Substrate Specificity |
Abstract | Oxaloacetate (OAA) decarboxylase (E.C. 4.1.1.3) 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. |
DOI | 10.1007/BF00871217 |
Alternate Journal | Antonie Van Leeuwenhoek |
Citation Key | 193 |
PubMed ID | 7771770 |