Biosynthesis of ethyl butyrate by immobilized recombinant Rhizopus oryzae lipase expressed in Pichia pastoris

A recombinant Rhizopus oryzae lipase expressed in Pichia pastoris has been immobilized in three different kinds of supports: EP100, Eupergit®CM and Octadecyl-Sepabeads. These immobilized derivatives have been used to catalyze the synthesis of ethyl butyrate. The effect of different parameters on initial reaction rate and yield has been studied for each biocatalyst. The variables selected for the study were: temperature, water content, shaking speed, enzyme loading, type of solvent and substrates molar ratio. The results showed the EP100 derivative as the best choice in terms of initial rate and yield but, when the residual activity as well as the reutilization capacity was studied, Octadecyl-Sepabeads exhibits the highest stability, so it was finally chosen as the best biocatalyst.

► Recombinant Rhizopus oryzae lipase (rROL) catalyzes the synthesis of ethyl butyrate.
► Determination of the influence of different parameters on synthesis reaction.
► rROL immobilized on EP100 showed the highest initial synthesis rate.
► rROL immobilized on Sepabeads showed the best operational stability.