Purification by affinity chromatography of 2,4-dienoyl-CoA reductases from bovine liver and Escherichia coli.

摘要: 1 Dye-ligand chromatography using immobilized Cibacron blue F3GA (blue Sepharose CL-6B) and Procion red HE3B (Matrex gel A) as matrices general ligand employing 2′,5′-ADP (2′,5′-ADP- 4B) 3′,5′-ADP (3′,5′-ADP- Agarose) were employed for purification of NADPH-dependent 2-enoyl-CoA reductase 2,4-dienoyl-CoA from bovine liver (formerly called 4-enoyl-CoA [Kunau, W. H. Dommes, P. (1978) Eur. J. Biochem. 91, 533–544], well Escherichia coli. 2 The mitochondria was separated by dye-ligand A/KCl gradient) affinity (2′,5′-ADP - 4B/NADP gradient). The enzyme obtained in a highly purified form. 3 The to homogeneity CL-6B, Matrex A, 2′,5′-ADP–Sepharose 4B chromatography. 4 The bacterial completely ion-exchange on DEAE-cellulose followed single step biospecific elution the column with substrate, trans, trans-2,4-decadienoyl-CoA. 5 The application reductases taking part β-oxidation unsaturated fatty acids is discussed. It concluded that making use coenzyme specificity binding substrate essential obtaining homogeneous preparations.