title: Antibacterial Peptide CecropinB2 Production via Various Host and Construct Systems.
abstract: Cecropin is a cationic antibacterial peptide composed of 35-39 residues. This peptide has been identified as possessing strong antibacterial activity and low toxicity against eukaryotic cells, and it has been claimed that some types of the cecropin family of peptides are capable of killing cancer cells. In this study, the host effect of cloning antibacterial peptide cecropinB2 was investigated. Three different host expression systems were chosen, i.e., Escherichia coli, Bacillus subtilis and Pichia pastoris. Two gene constructs, cecropinB2 (cecB2) and intein-cecropinB2 (INT-cecB2), were applied. Signal peptide and propeptide from Armigeres subalbatus were also attached to the gene construct. The results showed that the best host for cloning cecropinB2 was P. pastoris SMD1168 harboring the gene of pGAPzαC-prepro-cecB2 via Western blot confirmation. The cecropinB2 that was purified using immobilized-metal affinity chromatography resin showed strong antibacterial activity against the Gram-negative strains, including the multi-drug-resistant bacteria Acinetobacter baumannii.
authors.affiliation: Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan. s099065033@gmail.com.
authors.forename: Wei-Shiang
authors.initials: WS
authors.lastname: Lai
authors.affiliation: Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan. sherrykan0402@gmail.com.
authors.forename: Shu-Chen
authors.initials: SC
authors.lastname: Kan
authors.affiliation: Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan. teddy@ms23.url.com.tw.
authors.forename: Chia-Chi
authors.initials: CC
authors.lastname: Lin
authors.affiliation: Biotechnology Center, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan. cjshieh@nchu.edu.tw.
authors.forename: Chwen-Jen
authors.initials: CJ
authors.lastname: Shieh
authors.affiliation: Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan. young123tw@gmail.com.
authors.forename: Yung-Chuan
authors.initials: YC
authors.lastname: Liu
mesh_terms: D040981:Acinetobacter baumannii / Q000187:drug effects / Q000254:growth & development; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D047668:Inteins / Q000235:genetics; D008826:Microbial Sensitivity Tests; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology
chemical_list: D023181:Antimicrobial Cationic Peptides; D019476:Insect Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C031162:cecropin B protein, Insecta
keywords: Bacillus subtilis; Pichia pastoris; antibacterial peptides; cecropinB2; multi-drug-resistant