Isolation and biochemical characterisation of a bacteriocin-like substance produced by Bacillus amyloliquefaciens An6

Abstract

This study focuses on the isolation and characterisation of a peptide with bacteriocin-like properties from Bacillus amyloliquefaciens An6. Incubation conditions were optimised, and the effects of the incubation period and of carbon and nitrogen sources were investigated. The produced bacteriocin was partially purified with ammonium sulphate precipitation, dialysis and ultrafiltration and was then biochemically characterised. Maximum bacteriocin production was achieved after 48h of incubation in a culture medium containing 20g/L starch and 10g/L yeast extract, with an initial pH 8.0 at 30°C under continuous agitation at 200rpm. The bacteriocin was sequentially purified and its molecular weight was determined to be 11kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The bacteriocin was relatively heat-resistant and was not sensitive to acid and alkaline conditions (pH 4.0-10.0). Its inhibitory activity was sensitive to proteinase K but was resistant to the proteolytic action of alcalase, trypsin, chymotrypsin and pepsin. In conclusion, bacteriocin An6, owing its wide spectrum of activity as well as its high tolerance to acidic and alkaline pH values, temperature and proteases shows great potential for use as a food biopreservative.

Keywords: Antimicrobial activity; Bacillus amyloliquefaciens; Bacteriocin; Biochemical characterisation.