In order to obtain the strains of high a-amylase activity, 200 strains were obtained through isolation, the crude enzyme activity of 20 of the strains were higher than the others through the Yoo YJ’s method. The strain L1-2, which had a highest enzyme activity, reaching 766 U/mL, was Bacillus amyloliquefaciens identified by 16S rRNA. The a-amylase gene was cloned and inserted into the pET32a vector, then transformed into E. coli BL21 (ED3). The enzyme activity of recombinant protein was 1747.20 U/mg. The optimal pH was 6, and it remained more than 80% activity between 5 and 7. The optimal temperature was 60° C, and there remained more than 80% activity between 40°C and 70°C. The recombinant protein was significantly activated by Ca²⁺, significantly inhibited by Cu²⁺; Co²⁺; Fe³⁺; K⁺. The study was the genetically modification foundation of the a-amylase production strains and meeting the industrial production needs.