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Staphylococcal lipases
Author(s) -
Vadehra D. V.
Publication year - 1974
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02532687
Subject(s) - chemistry , lipase , chromatography , hydrolysis , triacylglycerol lipase , oleic acid , tributyrin , stearic acid , palmitic acid , biochemistry , fatty acid , organic chemistry , enzyme
A lipase rich fraction was isolated from the cell free supernatant of 24 hr broth culture of Staphylococcus aureus B‐120, grown in trypticase soy broth at 37 C. Lipase from the cell free supernatant was precipitated with equal volumes of absolute ethanol. This fraction was purified further by differential precipitation at pH 8.6 and 4.3. Subsequent purification, using Sephadex G‐200 and BioGel 300, yielded a preparation with 350–450‐fold increase in specific activity. The purified lipase had an optimum pH of 8.5 at 37 C. The electrophoretic mobility was‐7.78×10 −5 cm 2 /volt/sec. The sedimentation coefficient for the two peaks was 2.85 and 8.5, respectively, and the mol wt was 100,000. The purified lipase hydrolyzed a variety of natural oils and fats. The amount of free fatty acids liberated from hydrogenated soybean oil (iodine value<3) was one‐third compared to natural oils and fats. Gas chromatographic analysis of hydrolyzed synthetic triglyceride, with palmitic, stearic, and oleic acids at the rac 1, 2, and 3 positions, respectively, indicated that the enzyme was capable of hydrolyzing the glycerolfatty acid bonds at all three positions. The yield was 40% palmitic, 20% stearic, and 39% oleic acids. Formaldehyde, mercaptoethanol, cysteine, glutathione, and terramycin had inhibitory effects upon lipase activity while hydrogen peroxide, streptomycin, and sodium taurocholate had a stimulatory effect upon the activity.