title: Expression of a novel bacteriocin-the plantaricin Pln1-in Escherichia coli and its functional analysis.
abstract: A potential bacteriocin gene was isolated from 18575 ORFs by bioinformatics methods. It was named pln1, and cloned into pET32a. Then, it was expressed as a thioredoxin-Pln1 fusion protein in Escherichia coli BL21 (DE3). The fusion protein was purified by Ni-NTA, and thioredoxin was removed by enterokinase. Finally, Pln1 was purified using a cation affinity column. The yields of fused and cleaved Pln1 peptides were 100-110 mg/l and 9-11 mg/l, respectively. Pln1 was stable in an acidic environment and at temperatures below 60 °C, but was easily degraded under alkaline conditions and by protease treatment. The cleaved and purified Pln1 showed strong antimicrobial activity against gram-positive bacteria such as Micrococcus luteus CMCC 63202, Staphylococcus epidermidis, Lactococcus lactis NZ3900, Lactobacillus paracasei CICC 20241, and Listeria innocua CICC 10417. In particular, Pln1 had a better activity against methicillin-resistant S. epidermidis (MRSE) than nisin, thereby offering an attractive approach to counter bacterial antibiotic resistance.
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: mfq2012@outlook.com.
authors.forename: Fanqiang
authors.initials: F
authors.lastname: Meng
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: zhaohz@njau.edu.cn.
authors.forename: Haizhen
authors.initials: H
authors.lastname: Zhao
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: zhangchong@njau.edu.cn.
authors.forename: Chong
authors.initials: C
authors.lastname: Zhang
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: lufengxia@njau.edu.cn.
authors.forename: Fengxia
authors.initials: F
authors.lastname: Lu
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: bxm43@njau.edu.cn.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Bie
authors.affiliation: College of Food Science and Technology, Nanjing Agriculture University Nanjing, 210095, China. Electronic address: fmb@njau.edu.cn.
authors.forename: Zhaoxin
authors.initials: Z
authors.lastname: Lu
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli; D015870:Gene Expression; D005798:Genes, Bacterial; D048191:Lactiplantibacillus plantarum / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D011994:Recombinant Proteins
keywords: Antimicrobial activity; Heterologous expression; MRSE; Plantaricin