Enhanced Activity, Enantioselectivity and Stability of Papain in Asymmetric Hydrolysis of D , L - p -Hydroxyphenylglycine Methyl Ester with Ionic Liquid.

Papain-mediated asymmetric hydrolysis of D , L - p -hydroxyphenylglycine methyl ester ( D , L -HPGME) was examined in the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM·BF 4 ) and different solvents. The activity of the enzyme varied widely with change in BMIM·BF 4 concentration, with 12.5% (v/v) being the optimum BMIM·BF 4 concentration for the reaction. Papain displayed much higher hydrolytic activity and enantioselectivity in phosphate buffer solution of 12.5% (v/v) BMIM·BF 4 (pH 7.0) than in other media examined. Comparative studies on the kinetics and activation energy ( E a ) of this reaction performed in different media showed a higher V max , a lower K m and a lower E a for the reaction taking place in phosphate buffer solution of 12.5% (v/v) BMIM·BF 4 than in other media tested. The stability of papain at 45°C was considerably enhanced in BMIM·BF 4 as compared with aqueous buffer, 2-propanol and acetonitrile. A half-life time of 169 h was observed with BMIM·BF 4 in the presence of substrate, which was 9.2-16.8-fold higher than those with the other solvents. These results suggested that BMIM·BF 4 is an excellent reaction medium for this reaction. [ABSTRACT FROM AUTHOR]