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.
Wang
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Neurobiotechnology Center, Ohio State University, Columbus 43210.
Kolattukudy
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D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism*; D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004372:Ducks; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005990:Glycerol; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D012007:Rectum; D012627:Sebaceous Glands / Q000201:enzymology*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D012965:Sodium Chloride; D013379:Substrate Specificity
D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D009249:NADP; D000445:Aldehyde Oxidoreductases; C021351:hexadecanal dehydrogenase (acylating); D005990:Glycerol