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Novel expression system for large-scale production and purification of recombinant class IIa bacteriocins and its application to piscicolin 126

Gibbs, G.M., Davidson, B.E., Hillier, A.J.
Applied and environmental microbiology 2004 v.70 no.6 pp. 3292-3297
Carnobacterium piscicola, bacteriocins, structural genes, molecular cloning, gene expression, Escherichia coli, recombinant proteins, protein synthesis, food preservatives, disulfide bonds, purification, antibacterial properties, potassium, membrane permeability, cell membranes, Listeria monocytogenes, Enterococcus faecalis, food biopreservatives
Piscicolin 126 is a class IIa bacteriocin isolated from Carnobacterium piscicola JG126 that exhibits strong activity against Listeria monocytogenes. The gene encoding mature piscicolin 126 (m-pisA) was cloned into an Escherichia coli expression system and expressed as a thioredoxin-piscicolin 126 fusion protein that was purified by affinity chromatography. Purified recombinant piscicolin 126 was obtained after CNBr cleavage of the fusion protein followed by reversed-phase chromatography. Recombinant piscicolin 126 contained a single disulfide bond and had a mass identical to that of native piscicolin 126. This novel bacteriocin expression system generated approximately 26 mg of purified bacteriocin from 1 liter of E. coli culture. The purified recombinant piscicolin 126 acted by disruption of the bacterial cell membrane.