Rational Development Strategies with N-Glycosylation Engineering of Rhizomucor miehei Lipase for Biodiesel Production

Abstract

The construction of methanol-resistant lipases with high catalytic activity is world-shattering for biodiesel production. A semi-rational method has been constructed to enhance the properties of lipase by introducing N-glycosylation sites in the loop structure. The enzyme activities of the mutants N288 and N142 increased to 56.09 and 41.56 times relative to that of wild type, respectively. After incubation in 50% methanol for 2.5 h, the residual activities of N314 and N174-1 were 95% and 85%, respectively. Additionally, the biodiesel yield of all mutants has improved after adding methanol once in 24 h. Among them, N288 increased the quantity of biodiesel from colza oil from 9.49% to 88%, and N314 increased the amount of biodiesel from waste soybean oil from 8.44% to 70%. This study provides an effective method to enhance the properties of lipase and improve its application potential in biodiesel production.

Keywords: N-glycosylation, Loop structure, Rhizomucor miehei lipase, Methanol tolerance, biodiesel