Development of a novel amphipathic peptide and coiled coil peptide-lipase fusion system to increase hydrophobic substrate accessibility and its biotechnological applications

Abstract

Lipases represent a versatile class of biocatalysts with numerous potential applications in industry, including the production of biodiesel via enzyme-catalyzed transesterification. Nevertheless, relatively slow enzyme reaction rates caused by poor interaction between the lipase and lipid hinder the utilization of lipases for industrial biodiesel production. In this article, we have investigated the performance of NKC-M37-MAT, a variant of Photobacterium lipolyticum M37 lipase engineered by fusion with an amphipathic peptide and a coiled-coil peptide. We tested its performance with a series of esters as well as triglycerides. Compared with wild-type M37 lipase, the NKC-M37-MAT lipase showed consistently higher catalytic activity-about 54-fold higher with an olive oil substrate-and substantially reduced the biodiesel production time from 30 to 6 h. Differences in the observed rates for wild-type M37 and NKC-M37-MAT are related to surface properties of the lipase and the conditions at the interface between the lipase and lipid substrates.