Solubilization, purification and characterization of fatty acyl-CoA reductase from duck uropygial gland.

Membrane-bound fatty acyl-CoA reductase from the uropygial gland of duck has been solubilized from the microsomal preparation with 20% glycerol and 3 M NaCl and purified to homogeneity by Blue A agarose and Palmitoyl-CoA agarose affinity column chromatography followed by Suprose-6 gel filtration. The molecular mass of the enzyme was estimated by SDS-PAGE to be 56 kDa. The enzyme was stable in the presence of 20% glycerol and 1M NaCl and required NADPH for activity. The apparent Kms of the purified enzyme for palmitoyl-CoA and NADPH were 29 microM and 67 microM, respectively. The enzyme activity could be enhanced by the addition of lipid, and the presence of 2 mg/ml BSA enhanced the reductase activity by 5-fold.