Kinetic and Transesterification Properties of Lipase from Sprouted Melon (Cucumeropsis manni) Seeds
Crude lipase (acetone powder) was extracted from freshly sprouted melon seeds (Cucumeropsis manni). The activity, kinetic properties (effect of time, pH, and enzyme and substrate concentration, respectively) as well as the ability of the crude lipase to catalyze the production of methyl esters (biodiesel) were examined. The enzyme activity was determined using n-hexane as the solvent (1:2 v/w solvent: substrate ratio) and the transesterification product was analyzed by HPLC. A linear relationship was observed between reaction time and rate of lipolysis with the optimal activity at 2hr of incubation. Furthermore, the lipase was optimally active at acid pH 5 and lipolysis was achieved optimally when the amount of enzyme was 2.0g. Rate of lipolysis was observed to increase linearly at concentrations up to 5.0g of substrate above which a drop in the rate, with no apparent decrease in activity, was observed. The Km (6.25g) and Vmax (13.33%FFA/hr) were also determined. Analysis of the transesterification product yielded 0.61% alkyl ester, 0.81 %FFA, 93.17% TAG, 4.15% 1, 3-DAG and 1.26% 1, 2-DAG while transesterification efficiency was determined to be at 0.588%. Biodiesel (alkyl esters) prepared with the crude lipase was had a density of 0.872 g/mL while its cloud and pour points were 22°C and 12°C, respectively. The results from this research showed that an active lipase was isolated from sprouted melon seeds. However, the fuel properties of the biodiesel produced did not meet international transportation fuel standards. In order to be used industrially, better reaction conditions need to be established for the lipase.