title: Expression and purification of a fusion-typed pediocin PA-1 in Escherichia coli and recovery of biologically active pediocin PA-1.
abstract: Pediocin PA-1 is a representative class IIa bacteriocin which is small and heat-stable and has a consensus motif, -YGNGV-. The plasmid pQE40PED, encoding pediocin PA-1 fused with His-tagged mouse dihydrofolate reductase (DHFR), was constructed and introduced into Escherichia coli M15 strain. The fusion protein was overexpressed in the strain after induction of isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography. For the recovery of biologically active pediocin PA-1, the purified fusion protein was cleaved by Factor Xa protease and the liberated pediocin PA-1 was finally purified by ultrafiltration with a 75% yield. The molecular mass of the purified recombinant pediocin PA-1 was the same as that of native pediocin PA-1 on an electrophoresis gel.
authors.affiliation: Food Safety Research Division, Korea Food Research Institute, Seongnam, Gyeonggi 463-746, Korea.
authors.forename: Gi-Seong
authors.initials: GS
authors.lastname: Moon
authors.forename: Yu-Ryang
authors.initials: YR
authors.lastname: Pyun
authors.forename: Wang June
authors.initials: WJ
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005516:Food Microbiology; D015870:Gene Expression; D008969:Molecular Sequence Data; D008970:Molecular Weight; D000070017:Pediocins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins
chemical_list: D001430:Bacteriocins; D000070017:Pediocins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C074288:pediocin PA-1