Biotechnology and Bioengineering, Vol.43, No.11, 1081-1086, 1994
Solvation of Cbz-Amino Acid Nitrophenyl Eaters in Organic Media and the Kinetics of Their Transesterification by Subtilisin
Subtilisin Carlsberg adsorbed on silica particles has been used to catalyze the transesterification of CBZ-Ala-ONp and CBZ-Leu-ONp with 1-butanol in organic systems preequilibrated to water activity of 0.93. Initial reaction rates are conveniently followed by extraction of the released nitrophenol into an alkaline aqueous phase. Kinetic parameters were determined for varied ester concentrations in toluene, isopropyl ether, and hexane. The effect of solvent on substrate solvation was determined by solubility measurements. Much of the observed effect of solvent on V-m/K-m may be accounted for by solvation differences. The residual effect of solvent on K-m, after discounting solvation differences, is completely opposite to the apparent trend.