Enzyme Design for Nonaqueous Media: Optimization of Enzymatic Catalysis forOrganic Solvent Systems

摘要

From a biotechnological perspective there are many potential advantages ofemploying enzymes in organic as opposed to aqueous media. To that end, we have concluded an initial three-year research program in the area of enzymatic catalysis in organic solvents. Our studies have focused on the effect of protein hydration on subtilisins BPN' and Carlsberg in nonaqueous media. Investigations on protein engineered mutants, catalyst engineering studies, and structural studies, primarily employing EPR spectroscopy, have revealed fundamental information on the role of water, the the nature of enzyme structure, and the effects of solvents on the catalytic activity in organic solvents. This study has also resulted in several rational methods to dramatically improve enzyme activity under anhydrous conditions. Catalytic activities in organic solvents can now be expressed at levels similar to that in water--this is a major advance in the field of nonaqueous enzymology. (jg).