Antimicrobial Activity of Subtilin from Bacillus Subtilis JDB-1 and Cloning spaS Gene

Abstract

Abstract: The bacterial subtilin was purified from the fermentation broth of Bacillus subtilis JDB-1 by using the ammonium sulfate precipitation,and its antimicrobial activity against five tested microorganisms was investigated by using the agar diffusion test or the paper disc test.The spaS gene encoding subtilin was amplified from genomic DNA of Bacillus subtilis JDB-1 by PCR，and the PCR products cloned into the pMD18-T vector and subsequently introduced the competent cells of E. coli DH5α.The recombinant plasmid pMD18-spaS was identified by PCR and then sequenced.The results showed that the purified subtilin displayed antimicrobial activities against E. coli K12,Pseudomonas putida AS1.1003,Gibberella moniliformis JSD-8 and Candida albicans ATCC10123,respectively.The sequence analysis showed that the coding region spaS gene of Bacillus subtilis JDB-1 was 171bp in length,and the nucleotide sequence similarity of the spaS genes between the JDB-1 strain and the previously reported other bacterial strains was 95%.

Key words: bacillus subtilis, subtilin, antimicrobial activity, cloning, sequencing

WU Rui-Fang, GONG Feng-Juan, YANG Yi-Man, EnTe-Ma-Ke-.Bu-La-Ti-Bai . Antimicrobial Activity of Subtilin from Bacillus Subtilis JDB-1 and Cloning spaS Gene[J]. journal6, 2011, 32(4): 92-98.

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