Lipase from Pseudomonas fragi CRDA 323: partial purification, characterization and interesterification of butter fat.
Several strains of Pseudomonas were screened for lipase production using the rhodamine agar diffusion test. On the basis of the diameter of the halo produced, P. fragi CRDA 323 and P. putida ATCC 795 were considered to be good and weak lipase producers respectively. P. fragi, cultured in a 2-l fermenter, produced a maximal amount of lipase after 3-4 days of incubation at 27 degrees C. The lipase extract of P. fragi was obtained by acidification of culture supernatant at pH 4.0 and partially purified with ammonium sulphate precipitation. The majority of lipase activity (42%) was located in fraction IV, precipitated at 20%-40% of saturation, with a 19-fold enzyme purification. The Km and Vmax values for the partially purified enzymatic extract (fraction IV) were 0.70 mg/ml and 0.97 x 10(-3) U/min respectively. Fraction IV, which showed an optimum activity at pH 8.5, was used for the interesterification of butter fat in a microemulsion free co-surfactant system containing Span 60 (sorbitol monostearate) and Tween 60 (polyoxyethylene sorbitan monostearate) in the ratio 48:52 (v/v). The results showed that the interesterification of butter fat resulted in a considerable decrease in long-chain saturated fatty acids (C12:0, C14:0 and C16:0) with a concomitant increase in C18:0 and C18:1 at the sn-2 position of selected triacylglycerols. In addition, the results demonstrated an increase in the fatty acids (C12:0, C14:0 and 16:0) among the 1 and 3 positions of the triacylglycerol molecules of modified buffer fat accompanied by a decrease in C18:0 and C18:1.
Pabai
F
F
Department of Food Science and Agricultural Chemistry, McGill University, Ste Anne de Bellevue, Québec, Canada.
Kermasha
S
S
Morin
A
A
D002079:Butter; D004591:Electrophoresis, Polyacrylamide Gel; D004952:Esters / Q000378:metabolism; D005223:Fats / Q000378:metabolism*; D005227:Fatty Acids / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000302:isolation & purification*; D011549:Pseudomonas / Q000201:enzymology*
D004952:Esters; D005223:Fats; D005227:Fatty Acids; D002079:Butter; D008049:Lipase