α-Chymotrypsin shows higher activity in water as well as organic solvents after three phase partitioning

Alpha-Chymotrypsin was found to show a 119% increase in activity after three phase partitioning. The k_cat/K_m of the partitioned enzyme (TPP-C) for hydrolysis of Bz-Tyr-OEt in aqueous medium at 25°C was found to be 48.3×10⁴ mM^-1 min^-1 as compared to the corresponding value of 17.7×10⁴ mM^-1 min^-1 for the untreated control (C). The λ_max of the fluorescence emission spectrum of TPP-C showed 178% increase in the quantum yield when compared to C. TPP-C showed a 2.94 and 3.58 fold increase (as compared to C) in initial rates for formation of the ester Ac-Phe-OEt (from Ac-Phe and ethanol) in low water containing toluene and n-octane, respectively. It was found that TPP-C also showed the phenomenon of pH memory. At 5% (v v^-1) water (in t-amyl alcohol), while no esterification was observed with C, TPP-C still showed significant level of esterification activity.