Novel expression system for large-scale production and purification of recombinant class IIa bacteriocins and its application to piscicolin 126.
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.
Gibbs
Gerard M
GM
Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria 3052, Australia.
Davidson
Barrie E
BE
Hillier
Alan J
AJ
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D007777:Lactobacillaceae / Q000235:genetics* / Q000378:metabolism; D008089:Listeria monocytogenes / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D001430:Bacteriocins; D011994:Recombinant Proteins; C470824:piscicolin 126; D013879:Thioredoxins