Screening, Identification, Cloning, Expression and Characterization of alpha-amylase from Bacillus amyloliquefaciensAU Lv, Xin (geneium@scu.edu.cn) Zhu, Weihua Cao, Yi

摘要

In order to obtain the strains of high alpha-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 alpha-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 degrees C, and there remained more than 80% activity between 40 degrees C and 70 degrees 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 alpha-amylase production strains and meeting the industrial production needs.C1 Cao, Yi; Sichuan Univ, Coll Life Sci, Chengdu, Peoples R ChinaSC Biochemistry & Molecular Biology; Genetics & Heredity