Characterization of lipase of Pseudomonas fluorescens 27 based on fatty acid profiles.
The objectives of this research were to isolate lipase from Pseudomonas fluorescens 27, to compare the purity of the partially purified lipase preparation with crude extract, and to determine if bands of lipase activity revealed by disc gel electrophoresis liberated different free fatty acids from milk fat. Lipases were isolated from a shaken skim milk culture of P. fluorescens 27 by using ion-exchange chromatography on DEAE cellulose (Whatman DE 32) and gel filtration on Sephadex G-150. The principal lipase-rich fractions from gel filtration represented 6.2% of total lipolytic activity. Disc gel electrophoresis of partially purified enzyme revealed two protein bands. These protein bands were cut from disc electrophoresis gels and used as an enzyme source for reaction with butter oil. Free fatty acids were isolated from the assay mixture, separated, and quantified by gas chromatography. Data from gas chromatographic analysis indicated that P. fluorescens 27 produces at least two different lipases.
D000818:Animals; D002417:Cattle; D005223:Fats; D005227:Fatty Acids / Q000032:analysis*; D066298:In Vitro Techniques; D008049:Lipase / Q000032:analysis / Q000302:isolation & purification*; D008066:Lipolysis; D008892:Milk; D011551:Pseudomonas fluorescens / Q000201:enzymology*
D005223:Fats; D005227:Fatty Acids; D008049:Lipase