Sterol ester hydrolase in Fusarium oxysporum.
Two electrophoretically different forms of sterol ester hydrolase (EC 3.1.1.13) were obtained from the cytoplasmic extract of the mycelia of Fusarium oxysporum. The entities, estimated at 60,000 (I) and 15,000 (II) molecular weights, were obtained in Sephadex G100 column chromatography of the ammonium sulfate precipitate from the cytoplasmic extract. A third form III, 75,000 MW, was obtained from the culture filtrate. The activity of the enzyme was increased by Triton X-100 and was not inhibited by p-chloromercuribenzoate (PCMB), a sulfhydryl reagent. The enzymes I and II were inhibited differentially by NaCl. The optimal activities of forms I, II and III occurred at pH 4.8, pH 8.0 and pH 7.0, respectively. The apparent Km values of 7.7 X 10(-5), 8.3 X 10(-5) and 10.5 X 10(-5), respectively, indicate a similar order of affinity for cholesteryl oleate at pH 7.1. The rate of hydrolysis of cholesteryl esters were in the order: linoleate greater than oleate greater than valerate greater than butyrate greater than acetate. Cholesteryl benzoate and palmitate were not hydrolyzed. The properties of the microbial enzyme are discussed in relation.
D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002729:Chloromercuribenzoates / Q000494:pharmacology; D005670:Fusarium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D012964:Sodium / Q000494:pharmacology; D002787:Sterol Esterase / Q000302:isolation & purification / Q000378:metabolism*; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D020245:p-Chloromercuribenzoic Acid
D002729:Chloromercuribenzoates; D046911:Macromolecular Substances; D020245:p-Chloromercuribenzoic Acid; D012964:Sodium; D002265:Carboxylic Ester Hydrolases; D002787:Sterol Esterase