Purification and characterization of oxalyl-coenzyme A decarboxylase from Oxalobacter formigenes

• Published in: Journal of Bacteriology Vol. 171; no. 5; pp. 2605 - 2608
• Main Authors: Baetz, A.L. (National Animal Disease Center, ARS, USDA, Ames, IA), Allison, M.J
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-05-1989
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; Acyl Coenzyme A - metabolism; BACTERIA; BACTERIE; Bacteriology; Biological and medical sciences; Carboxy-Lyases - isolation & purification; Carboxy-Lyases - metabolism; COENZIMAS; COENZYME; EPURATION; Fundamental and applied biological sciences. Psychology; Gram-Negative Anaerobic Bacteria - enzymology; Kinetics; LIASAS; LYASE; Metabolism. Enzymes; Microbiology; Molecular Weight; Oxalates - metabolism; Oxalic Acid; Oxalobacter formigenes; PROPIEDADES FISICO-QUIMICAS; PROPRIETE PHYSICO-CHIMIQUE; PURIFICACION; RUMEN; Spectrum Analysis; Substrate Specificity; Characterization; Purification; Enzyme; Oxalyl-CoA decarboxylase; Quaternary structure; Bacteria; Kinetic parameter; Physicochemical properties
• ISSN: 0021-9193; 1098-5530; 1067-8832
• DOI: 10.1128/jb.171.5.2605-2608.1989

Oxalyl-coenzyme A (oxalyl-CoA) decarboxylase was purified from Oxalobacter formigenes by high-pressure liquid chromatography with hydrophobic interaction chromatography, DEAE anion-exchange chromatography, and gel permeation chromatography. The enzyme is made up of four identical subunits (M(r), 65,000) to give the active enzyme (M(r), 260,000). The enzyme catalyzed the thiamine PP(i)-dependent decarboxylation of oxalyl-CoA to formate and carbon dioxide. Apparent K(m) and V(max) values, respectively, were 0.24 mM and 0.25 micromole/min for oxalyl-CoA and 1.1 pM and 0.14 micromole/min for thiamine pyrophosphate. The maximum specific activity was 13.5 microM oxalyl-CoA decarboxylated per min per mg of protein