Molecular characterization of transesterification activity of novel lipase family I.1.

Lipase's thermostability and organic solvent tolerance are two crucial properties that enable it to function as a biocatalyst. The present study examined the characteristics of two recombinant thermostable lipases (Lk2, Lk3) based on transesterification activity. Conversion of C12-C18 methyl ester with paranitrophenol was investigated in various organic solvent.

Cloning, heterologous expression, and characterization of a novel thioesterase from natural sample.

A novel thioesterse gene was successfully cloned and sequenced directly from natural sample of Domas Hot Spring, West Java, Indonesia. Homological analysis of the sequence showed that the gene appeared high homology to thioesterase genes with the highest to a putative thioesterase gene from uncultured at 66% identity. However, phylogenetic analysis showed that the protein was separated from the branch with other known thioesterases.

Effect of mutation at oxyanion hole residu (H110F) on activity of Lk4 lipase.

Mutant of lipase at oxyanion hole (H110 F) was constructed. The gene was highly expressed in BL21 (DE3) and the recombinant protein was purified using Ni-NTA affinity chromatography. The activity of mutant enzyme was significantly increased compared to that the wild type.