E156S and V165I mutation were introduced into subtilisin E gene by site-directed mutagenesis. The mutated gene fragments were recombined with pBE-2 which is a shuttle vector between E.coli and Bacillus subtilis. The recombinant plasmids were used to transform B.subtilis DB104，a mutant strain deficient in alkaline and neutral protease，then they were expressed. They were (M222A，E156S) and (M222A，E156S，V165I). The property analysis of these enzymes revealed that the Subtilisin E E156S substitution enhenced the hydrolysis K_cat/K_m by 90% while keeping thermal stability and oxidation-resistance unchanged，however the V165I mutation reduced the K_cat/K_m value.