Investigation of The Activity of Lipase Variants on Different 4-Nitrophenyl Esters by Spectrophotometric Assay

Microbial enzymes are important tools that frequently used in the field of biotechnology. These microbial enzymes, which play a role intracellular biological reactions, many different industries. Lipase, proteases and amylases members hydrolytic enzymes. Lipase enzyme, has most common usage area among is an enzyme catalyzes hydrolysis ester bonds between lipid molecules. The activity lipase commonly measured by spectrophotometric method. P-nitrophenol esters preferred for kinetic analysis. In analysis method, colored product as result p-nitrophenol substrates enzyme. this study, p-nitrophenyl acetate (Acetic acid 4-nitrophenyl ester), butyrate (Butyric octanoate (Octanoic ester, 4 -Nitrophenyl caprylate), dodecanoate (Dodecanoic palmitate (p-Nitrophenyl palmitate, Hexadecanoic ester) were used. products formed incubation with carbon lengths periods spectrophotometrically. Trials carried out at 25°C 5min-120min. As experiments four repetitions, it was determined varies according to length chain substrates. Vmax values wild calculated 0.42 U/mg protein, 0.95 1.1 0.78 0.18 protein pNP-acetate, pNP-buritate, pNP-octanoate, pNP-dodecanoate, pNP-palmitate, respectively. It on very low. eight-carbon pNP-octanoate substrate higher than other