Efficacious Extraction and Purification Technique of a Potential Antimycobacterial Bacteriocin Produced by Bacillus subtilis (MK733983) of Ethnomedicinal Origin.

Abstract

The rapid proliferation of antibiotic-resistant microbes has imposed an urgent need for development of novel antimicrobial agents with diverse mechanisms. This study reports a novel extraction method with salting-in and salting-out method for obtaining potential bacteriocin from Bacillus subtilis (MK733983) of ethnomedicinal origin. This technique extracted bacteriocin with desired antimicrobial peptide moieties that showed creditable minimum inhibitory concentrations, thermostability and efficacy compared to all other extraction protocols attempted. Further study used a unique scheme of steps in RP-HPLC purification process using methanol-water as solvents for the bacteriocin that achieved an outstanding antimicrobial activity against Staphylococcus aureus (MTCC 737). The bacteriocin is sensitive to proteases, confirming its proteinaceous nature and showed promising heat stability up to 70°C for 10min. Bacteriocin extracted from a series of ammonium sulphate precipitation showed MIC values 350µg and 300µg for Mycobacterium smegmatis and Staphylococcus aureus respectively. On the other hand, bacteriocin extracted by using chloroform showed MIC values 400µg and 300µg for M. smegmatis and Staphylococcus aureus. All the results implicate the efficacy of bacteriocin and future prospect as an effective antimicrobial agent.