In this study we explored the efficiency of the additive methyl-beta-cyclodextrin (MbetaCD) 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 colyophilization of subtilisin with MbetaPCD preserves the enzyme's active site tertiary structure rendering a highly active and enantioselective catalyst.