Enzyme activation in organic solvents: Co-lyophilization of subtilisin Carlsberg with methyl-β-cyclodextrin renders an enzyme catalyst more active than the cross-linked enzyme crystals

In this study we explored the efficiency of the additive methyl‐β‐cyclodextrin (MβCD) to enhance the activity and enantioselectivity of the serine protease subtilisin Carlsberg in organic solvents. These two parameters, measured for different transesterification reactions and in several solvents, are compared with results obtained by using two additional preparations of the same enzyme: lyophilized powder and cross‐linked enzyme crystals (CLEC). The results suggest that co‐lyophilization of subtilisin with MβCD preserves the enzyme's active site tertiary structure rendering a highly active and enantioselective catalyst. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 53–59, 2002; DOI 10.1002/bit.10182