title: Expression of mouse beta defensin 2 in escherichia coli and its broad-spectrum antimicrobial activity.
abstract: Mature mouse beta defensin 2 (mBD2) is a small cationic peptide with antimicrobial activity. Here we established a prokaryotic expression vector containing the cDNA of mature mBD2 fused with thioredoxin (TrxA), pET32a-mBD2. The vector was transformed into Escherichia Coli (E. coli) Rosseta-gami (2) for expression fusion protein. Under the optimization of fermentation parameters: induce with 0.6 mM isopropylthiogalactoside (IPTG) at 34°C in 2×YT medium and harvest at 6 h postinduction, fusion protein TrxA-mBD2 was high expressed in the soluble fraction (>95%). After cleaved fusion protein by enterokinase, soluble mature mBD2 was achieved 6 mg/L with a volumetric productivity. Purified recombinant mBD2 demonstrated clear broad-spectrum antimicrobial activity for fungi, bacteria and virus. The MIC of antibacterial activity of against Staphylococcus aureus was 50 μg/ml. The MIC of against Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans) was 12.5μg/ml and 25μg/ml, respectively. Also, the antimicrobial activity of mBD2 was effected by NaCl concentration. Additionally, mBD2 showed antiviral activity against influenza A virus (IAV), the protective rate for Madin-Darby canine kidney cells (MDCK) was 93.86% at the mBD2 concentration of 100 μg/ml. These works might provide a foundation for the following research on the mBD2 as therapeutic agent for medical microbes.
authors.affiliation: Chengdu Blood Centre , 610041 Chengdu , China ; Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University , 610041 Chengdu , China.
authors.forename: Tianxiang
authors.initials: T
authors.lastname: Gong
authors.forename: Wanyi
authors.initials: W
authors.lastname: Li
authors.forename: Yueling
authors.initials: Y
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Jiang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Feng
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Jiang
authors.forename: Mingyuan
authors.initials: M
authors.lastname: Li
keywords: Antimicrobial activity; Expression; Mouse beta defensin 2; Purification
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