A super lipase: Engineering lipase to increase substrate accessibility by fusion with amphipathic and coiled-coil peptides

Lipases have been utilized industrially to produce biodiesel, oleochemicals, and pharmaceuticals. Many efforts such as metagenomics, directed evolution and enzyme immobilization have been devoted to enhance the lipase activity. Here, we designed a recombinant lipase, NKC-M37-MAT, that was generated by incorporating an N-terminal amphipathic peptide (NKC) and a C-terminal coiled-coil peptide (MAT) into Photobacterium lipolyticum M37 lipase in order to further extend catalytic activity by increasing substrate accessibility. The activity of NKC-M37-MAT lipase with olive oil was higher by a factor of 54 compared with the wild-type, which reduced biodiesel production time from 30 h to 6 h. Differences in reaction rates between wild-type M37 and NKC-M37-MAT were related to lipase surface properties and interface conditions between a lipase and the substrate. This novel approach shows promise as a platform technology to increase lipase activity for producing industrial-scale yields of biodiesel as well as of biochemicals synthesized from hydrophobic substrates.