Resumo

Although lipases have numerous potential applications in food, pharmaceutical, oleochemical, paper and biofuels industries, their large scale use remains limited, mainly because of the its production high cost and difficult for small quantity marketing. Production improvement, characterization of activity scope and development of novel applications for new microbial lipases become essential to advance lipases industrial scale use. This study aim was to evaluate Colletotrichum gloeosporioides lipase production using liquid substrate fermentation and apply this lipase in esterification reactions in organic medium. Therefore, the filamentous fungus was subjected to fermentation by 96 h. C. gloeosporioides showed maximum enzyme activity (0.75 U / mL.min) 48 h after process beginning, setting this time as enzymatic production batch end. Specific activity (4.7), protein production (0.16 mg / mL) and biomass (5 g) were determined at 48 h fermentation time. The metabolic activity turned the medium alkaline and under these conditions lipase was secreted. Upon established of the best fermentation conditions, lipase production took place followed by centrifugation and cell mass removal. Supernatant containing extracellular enzymes was lyophilized, dissolved in n-heptane and applied in esterification reactions using para-nitrophenyl palmitate (pNPP) as substrate. Ethanol or methanol was used to the transesterification reactions. When lipases undergo pNPP ester cleavage, the methyl or ethyl palmitate ester (according to used alcohol) are synthesized and para-nitrophenol (pNP) is released and dosed to achieve reaction yields. C. gloeosporioides lipase showed esterification activity with ethanol (0.007 U / kg.min) and methanol (0.003 U / kg.min) showing market potential application, since there are still no commercially available lipases for biodiesel industrial production. Ability presented by C. gloeosporioides lipase to carry on esterification with ethanol is remarkable, once Brazil ethanol production is higher than methanol one.