Properties of triacylglycerol lipase in a mitochondrial fraction from baker's yeast (Saccharomyces cerevisiae).
A triacylglycerol lipase in a mitochondrial fraction isolated from yeast (Saccharomyces cerevisiae) has been characterized and the hydrolysis studied kinetically using an insoluble artificial triacylglycerol suspension. 1. The triacylglycerol was hydrolyzed almost completely to fatty acids and glycerol. The lipase activity was inhibited by potassium fluoride and the sodium salts of -chloride, -glycocholate and -pyrophosphate as well as by protamine sulfate but at concentrations much too high to indicate that the lipase is a non specific esterase or a lipoprotein lipase. Also parachloromercuribenzoate inhibited the lipase activity. Inhibitory effect of fatty acid was observed at concentrations above 1mM. This inhibition may provide a regulatory mechanism of the lipase in vivo. 2. On the day of isolation the lipase activity of intact mitochondria at pH 7.5 and 30 degrees C was 400 nmol free fatty acid -h-1 - mg-1 at a triacylglycerol concentration of 9.0 mM. Sonication of the mitochondria increased the activity 2-3 fold. Freezing of the mitochondria also activated the lipase and this activation was dependent upon the freezing method, the concentration of mitochondrial protein and the presence of bovine serum albumin. 3. The particulate nature of the assay system was illustrated by the observation that the apparent Km value of the lipase increased with the concentration of mitochondrial protein. For each protein concentration the lipase had two apparent Km values when the activity was assayed with intact mitochondria, but only one when  assayed with submitochondrial particles. At the same protein concentration the Km value for the latter was identical with the "low affinity" Km for the lipase in intact mitochondria.
D002729:Chloromercuribenzoates / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism; D005459:Fluorides / Q000494:pharmacology; D005615:Freezing; D005990:Glycerol / Q000378:metabolism; D006000:Glycocholic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D009829:Oleic Acids / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013010:Sonication; D014280:Triglycerides / Q000378:metabolism
D002729:Chloromercuribenzoates; D005227:Fatty Acids; D009829:Oleic Acids; D014280:Triglycerides; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D008049:Lipase; D006000:Glycocholic Acid; D005990:Glycerol; D005459:Fluorides