Simultaneous single-step purification of thiolase and NADP-dependent 3-hydroxybutyryl-CoA dehydrogenase from Clostridium kluyveri.
Thiolase and NADP-dependent 3-hydroxybutyryl-CoA dehydrogenase from Clostridium kluyveri were purified by ion-exchange chromatography to near homogeneity in a simultaneous, single-step procedure. The yield of both enzymes was greater than 80%. Thiolase was purified approximately 8-fold with sp act 115 units/mg, whereas 3-hydroxybutyryl-CoA dehydrogenase was purified 14-fold with sp act 292 units/mg. Isoelectric points of the enzymes are 7.7 for thiolase and 7.8 for 3-hydroxybutyryl-CoA dehydrogenase. Milligram quantities of each of these enzymes are readily obtained from this fatty acid-producing organism.
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000302:isolation & purification*; D000101:Acetyl-CoA C-Acetyltransferase / Q000302:isolation & purification*; D000123:Acetyltransferases / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing
D000596:Amino Acids; D015094:3-Hydroxyacyl CoA Dehydrogenases; C021388:3-hydroxybutyryl-CoA dehydrogenase; D000123:Acetyltransferases; D000101:Acetyl-CoA C-Acetyltransferase