Colletotrichum gloeosporioides lipase: Characterization and use in hydrolysis and esterifications

Abstract

Colletotrichum gloeosporioides was evaluated for its capacity to produce extracellular lipase. A crude enzyme preparation obtained after 48 h of fermentation reached 742 U/L of lipolytic activity. Estimated molecular weight of proteins responsible for this activity was about 18 kDa as determined by SDS-PAGE and zymogram analysis. Enzyme preparation showed optimum pH at 10 and stability at optimum temperature (37°C) for 5 h. It showed tolerance to a wide range of salts (NH 4 + , Mg +2 , Ca +2 , Mn +2 and Sn +2) and to some solvents (methanol, ethanol, isopropanol, 1-butanol, acetonitrile n-heptane and n-hexane) in different concentrations. The crude enzyme preparation was applied in hydrolysis reactions on different substrates (waste cooking soybean oil, cocoa butter and palm kernel oil) reaching high yields (87.6; 80.1; 74.9%, respectively). The preparation was lyophilized and it was applied in the synthesis of pineapple flavor, by esterification with butanol and butyric acid. C. gloeosporioides lipolytic enzymes synthesized butyl butyrate with 70% yield, in experiments carried out for 24 h using 1:1 acid/alcohol molar ratio in n-heptane medium. The lyophilized preparation was also able to perform transesterification of alcohols and p-nitrophenyl palmitate in organic medium (n-hexane), showing better activity when propanol was used (5.4.10-3 U/kg.min). This study pointed the potential of alkaline lipolytic enzyme produced by C. gloeosporioides in biotechnological industry.