Acyl-CoA reductase and acyl-CoA: fatty alcohol acyl transferase in the microsomal preparation from the bovine meibomian gland.
Biosynthesis of wax esters, one of the two major products of the meibomian gland, was found to be catalyzed mainly by the microsomes of the bovine meibomian gland. The microsomal preparation catalyzed hexadecanoyl-CoA reduction to hexadecanol without any accumulation of the aldehyde intermediate. Maximal rates of reduction occurred at pH 6.5 and required both NADH and NADPH; the latter alone gave considerable rates whereas NADH alone was ineffective. Exogenous hexadecanal reduction catalyzed by the same preparation showed a preference for NADH. The hexadecanoyl-CoA saturation pattern was slightly sigmoidal and concentrations higher than 125 microM inhibited reduction. The fatty alcohol generated from hexadecanoyl-CoA was found as free alcohol and as wax esters. Esterification of hexadecanol to wax esters catalyzed by the meibomian gland microsomal preparation required exogenous acyl-CoA or ATP and CoA and was not affected by exogenous cholesterol. Maximal rates of esterification were observed at neutral pH. Hexadecanoyl-CoA concentrations higher than 125 microM inhibited esterification. Hexadecanol showed a typical substrate saturation pattern with an apparent Km of 125 microM. Radio gas-liquid chromatography showed that, in the presence of exogenous hexadecanoyl-CoA, hexadecanol gave hexadecyl hexadecanoate whereas in the presence of ATP and CoA both C16 and C18 endogenous acids were used to esterify the alcohol. Consistent with the composition of the meibomian gland secretion, exogenous acyl-CoA longer than C14 and shorter than C20 gave maximal rates of esterification of hexadecanol.
Kolattukudy
P E
PE
Rogers
L
L
D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D015256:Coenzyme A-Transferases; D005143:Eyelids / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008537:Meibomian Glands / Q000201:enzymology* / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology* / Q000378:metabolism; D009243:NAD / Q000378:metabolism / Q000494:pharmacology; D009249:NADP / Q000378:metabolism / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D013466:Sulfurtransferases / Q000378:metabolism*; D013997:Time Factors
D009243:NAD; D009249:NADP; D000445:Aldehyde Oxidoreductases; C021351:hexadecanal dehydrogenase (acylating); D013466:Sulfurtransferases; D015256:Coenzyme A-Transferases; C031212:acyl-CoA transferase