Purification and characterization of oxalyl-coenzyme A decarboxylase from Oxalobacter formigenes.
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 (Mr, 65,000) to give the active enzyme (Mr, 260,000). The enzyme catalyzed the thiamine PPi-dependent decarboxylation of oxalyl-CoA to formate and carbon dioxide. Apparent Km and Vmax values, respectively, were 0.24 mM and 0.25 mumol/min for oxalyl-CoA and 1.1 pM and 0.14 mumol/min for thiamine pyrophosphate. The maximum specific activity was 13.5 microM oxalyl-CoA decarboxylated per min per mg of protein.
Baetz
A L
AL
National Animal Disease Center, U.S. Department of Agriculture, Ames, Iowa 50010.
Allison
M J
MJ
D000214:Acyl Coenzyme A / Q000378:metabolism; D002262:Carboxy-Lyases / Q000302:isolation & purification* / Q000378:metabolism; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology*; D007700:Kinetics; D008970:Molecular Weight; D010070:Oxalates / Q000378:metabolism*; D019815:Oxalic Acid; D013057:Spectrum Analysis; D013379:Substrate Specificity
D000214:Acyl Coenzyme A; D010070:Oxalates; D019815:Oxalic Acid; D002262:Carboxy-Lyases; C059418:oxalyl CoA decarboxylase