Purification and characterization of a bioluminescence-related fatty acyl esterase from Vibrio harveyi.

Vibrio harveyi extracts contain three polypeptides (32, 42, and 57 kDa) which are involved in long-chain aldehyde biosynthesis and can be labeled with 3H tetradecanoic acid (+ATP) and/or 3Htetradecanoyl-CoA. These proteins have been separated from other labeled bands by ammonium sulfate fractionation, and the 32-kDa polypeptide has been further purified to homogeneity by ion-exchange, gel filtration, and hydroxylapatite chromatography. In aqueous buffers at pH 7, the 32-kDa protein catalyzes the hydrolysis of tetradecanoyl-CoA at a low rate (0.01 mumol/min/mg) to form free fatty acids. The thioesterase rate is slightly increased by phosphate, which also protects the enzyme against inhibition by the sulfhydryl reagent N-ethylmaleimide. Acyl-CoA cleavage is dramatically stimulated (up to 100-fold) by certain organic solvents, in particular glycerol and ethylene glycol, with the fatty acyl group being transferred to the alcohol acceptors. These enzymatic properties may be related to the role of the 32-kDa esterase in generating fatty acids for subsequent use in the V. harveyi bioluminescent system.