7-64: Searching the penetration site and enhancing the stability of Candida antarctica Lipase B in the hydrophilic organic solvent by using molecular dynamic simulation

The production of biodiesel by enzymatic transesterification has attracted much attention for high purity product and enables easy separation. Candida antarctica lipase B is one of the most important enzyme in the biodiesel reaction. In the reaction, CALB shows low organic solvent stability because of methanol. In this study, we concerned about enhancing organic solvent stability in hydrophilic solvents. In water-miscible solvents, enzyme loses its stability due to deformation of enzyme surface by solvent penetration effect. Molecular dynamic simulation was proceeded to search how the enzyme actually motions in organic solvents. RMSD value of each residue was calculated and distinguished for searching penetration site. By RosettaDesign, target residues were predicted for further mutagenesis. After in vitro experiment of mutants, we obtained organic solvent stability enhanced CALB variants. The detail will be explained and discussed.