Expression of mouse beta defensin 2 in Escherichia coli and its broad-spectrum antimicrobial activity
Braz. j. microbiol
;
42(3): 1180-1187, July-Sept. 2011. ilus
Article
in English
| LILACS
| ID: lil-607553
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 34ºC in 2×YT medium and harvest at 6 h postinduction, fusion protein TrxA-mBD2 was high expressed in the soluble fraction (>95 percent). 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 percent 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.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Recombinant Fusion Proteins
/
Antimicrobial Cationic Peptides
/
Beta-Defensins
/
Escherichia coli
/
Isopropyl Thiogalactoside
Language:
English
Journal:
Braz. j. microbiol
Journal subject:
Microbiology
Year:
2011
Type:
Article
/
Project document
Affiliation country:
China
Institution/Affiliation country:
Chengdu Blood Centre/CN
/
Sichuan University/CN
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