Two-step biocatalytic process using lipase and whole cell catalysts for biodiesel production from unrefined jatropha oil

To avoid lipase deactivation by methanol in the enzymatic transesterification process, a two-step biocatalytic process for biodiesel production from unrefined jatropha oil was developed. Unrefined jatropha oil was first hydrolyzed to free fatty acids (FFAs) by the commercial enzyme Candida rugosa lipase. The maximum yield achieved of FFAs 90.3 % at 40 A degrees C, water/oil ratio 0.75:1 (v/v), lipase content 2 % (w/w) after 8 h reaction. After hydrolysis, the FFAs were separated and converted to biodiesel by using Rhizopus oryzae IFO4697 cells immobilized within biomass support particles as a whole-cell biocatalyst. Molecular sieves (3 ) were added to the esterification reaction mixture to remove the byproduct water. The maximum fatty acid methyl ester yield reached 88.6 % at 35 A degrees C, molar ratio of methanol to FFAs 1.2:1, molecular sieves (3 ) content 60 % (w/w) after 42 h. In addition, both C. rugosa lipase and R. oryzae whole cell catalyst in the process showed excellent reusability, retaining 89 and 79 % yields, respectively, even after six batches of reactions. This novel process, combining the advantages of enzyme and whole cell catalysts, saved the consumption of commercial enzyme and avoid enzyme deactivation by methanol.