A simple and sensitive method for the estimation of microbial lipase activity.
A simple and sensitive method for the estimation of microbial lipase activity is described. In this method, lipase (EC 3.1.1.3) is incubated with an emulsified substrate and the fatty acid formed is estimated by high-performance liquid chromatography. The emulsified substrate, triolein, is prepared with various solubilizers, bovine serum albumin, gelatin, ovalbumin, gum arabic, Triton X-100, and n-octyl-glucopyranoside, in suitable buffers. The oleic acid, one of the products formed in the reaction, is separated on an ODS column with a premixed acetone-acetonitrile solvent system. Results with various lipase activities showed gum arabic to be the best among various solubilizers used to prepare the emulsified substrate. Also the effects of mono- and divalent ions on microbial lipase activities are analyzed. Finally, this method is compared with two other methods, titration and reversed micelles, and found to be simple and more sensitive.
Veeraragavan
K
K
Biochemical Engineering Section, National Research Council of Canada, Montreal, Quebec.
D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D004655:Emulsions; D005656:Fungal Proteins / Q000378:metabolism*; D006170:Gum Arabic; D008049:Lipase / Q000378:metabolism*; D011506:Proteins / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology; D013379:Substrate Specificity; D014304:Triolein / Q000378:metabolism
D002021:Buffers; D004655:Emulsions; D005656:Fungal Proteins; D011506:Proteins; D012997:Solvents; D014304:Triolein; D006170:Gum Arabic; D008049:Lipase