title: Expression of antimicrobial peptide LH multimers in Escherichia coli C43(DE3).
abstract: The tandem repeats of LFB15(W4,10)-HP(4-16) (LH) gene were cloned into vector pET32a(+) for recombinant expression in Escherichia coli. The E. coli C43(DE3) was successfully used as the expression host to avoid the cell death during induction in E. coli BL21(DE3). Fusion LH dimer was expressed as inclusion body at a portion of 35% of total cell protein and could be well purified by Ni(2+)-chelating chromatography. The recombinant LH was released by the cleavage of 50% formic acid, and its yield reached 11.3 mg/l with purity of 95%. The MIC(50) of 3.6 and 1.9 microM of recombinant LH against E. coli CMCC 44102 and Bacillus subtilis ATCC 6633 were determined, respectively. The results demonstrated that expression of tandem LH gene in E. coli C43(DE3) and formic acid cleavage would provide a potent efficient platform for the production of interested peptides.
authors.affiliation: Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun Nandajie St., Haidian District, Beijing 100081, People's Republic of China.
authors.forename: Zi-gang
authors.initials: ZG
authors.lastname: Tian
authors.forename: Tian-tang
authors.initials: TT
authors.lastname: Dong
authors.forename: Ya-lin
authors.initials: YL
authors.lastname: Yang
authors.forename: Da
authors.initials: D
authors.lastname: Teng
authors.forename: Jian-hua
authors.initials: JH
authors.lastname: Wang
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D001412:Bacillus subtilis / Q000187:drug effects; D001483:Base Sequence; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D005561:Formates / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D023181:Antimicrobial Cationic Peptides; D005561:Formates; D011993:Recombinant Fusion Proteins; C030544:formic acid