A theoretical analysis on comparing the enzyme performance in a single-step desymmetrization, single-step kinetic resolution, and two-step desymmetrization (i.e. a single-step desymmetrization followed by a sequent kinetic resolution) is reported. On the basis of ee* >= 0.95, a sufficient condition of E3E2-1 >= 10 and E-1 >= 2 is proposed for obtaining an acceptable yield of X-2R > 0.412 for the desired enantiomer in the two-step desymmetrization process, in comparison with E-1 >= 39 for the single-step desymmetrization and E3E2-1 20 for the single-step kinetic resolution. With the CALB-catalyzed hydrolytic desymmetrization of dipyrazolidyl3-phenylglutarate (1) in MTBE as the model system, enantiomerically pure (R)-monopyrazolidyl 3-phenylglutarate ((R)-2) can then bfdee prepared. Moreover from the kinetic analysis, the best reaction condition of using 20% water-saturated MTBE as the medium at 45 degrees C is selected for improving the enzyme activity and stereoselectivity. The thermodynamic analysis also indicates that the enzyme stereo-discrimination in the desymmetrization and sequent kinetic resolution is mainly entropic-driven. (C) 2015 Elsevier Ltd. All rights reserved.