Simultaneous single-step purification of thiolase and NADP-dependent 3-hydroxybutyryl-CoA dehydrogenase from Clostridium kluyveri

✍ Scribed by Mark X. Sliwkowski; Maris G.N. Hartmanis

• Book ID: 102986049
• Publisher: Elsevier Science
• Year: 1984
• Tongue: English
• Weight: 795 KB
• Volume: 141
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(84)90052-6

✦ Synopsis

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.