Purification and properties of sterol-ester hydrolase from Saccharomyces cerevisiae.
Sterol-ester hydrolase [EC 3.1.1.13] from Saccharomyces cerevisiae grown aerobically was solubilized with 1% Tween 20 and purified about 700-fold by the protamine sulfate treatment, DEAE-cellulose-, Sepharose 6B- and DEAE-cellulose column chromatographies. The molecular weight of the enzyme was estimated to be 70,000 by Sepharose 6B gel filtration. The enzyme activity showed two peaks of pH optimum at 4.4 and 6.8. Triton X-100 stimulated the activity as its low concentrations at both pH regions, but decreased the activity at its high concentrations at pH 6.8. The presence of Tween 20 or Tween 80 also stimulated the activity. These results were different from those in the previous report showing no stimulation of the crude enzyme by these detergents. The stimulation of the activity by phosphatidylcholine or low concentrations of lysophosphatidylcholine was similar to that by Triton X-100, and taurocholate was less effective than Triton X-100. The enzyme activity was inhibited by divalent cations such as Hg2+ and Cu2+.
D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008970:Molecular Weight; D017830:Octoxynol; D010713:Phosphatidylcholines / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011136:Polysorbates / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D002787:Sterol Esterase / Q000302:isolation & purification* / Q000378:metabolism; D013656:Taurocholic Acid / Q000494:pharmacology
D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D011136:Polysorbates; D011092:Polyethylene Glycols; D013656:Taurocholic Acid; D017830:Octoxynol; D002265:Carboxylic Ester Hydrolases; D002787:Sterol Esterase