Tandem repeat mhBD2 gene enhance the soluble fusion expression of hBD2 in Escherichia coli.
Human beta-defensin-2 (hBD2) is a cysteine-rich cationic antimicrobial peptide with low molecular weight that exhibits a broad range of antimicrobial activity. To improve the expression level of hBD2 in Escherichia coli, tandem repeats of mature hBD2 gene were constructed and expressed as fusion proteins (TrxA-nmhBD2, n = 1, 2, 4, 8) by constructing the vectors of pET32-nsmhBD2 (n = 1, 2, 4, 8). The results showed that the tandem repeats of mhBD2 gene were highly expressed in our constructed system. Comparing the expression levels of soluble mhBD2, BL21(DE3)/pET32-2smhBD2 was selected as an ideal recombinant strain for mature hBD2 production. Under the optimized conditions of cultivation and isopropylthiogalactoside (IPTG) induction, the maximum expression level of soluble mature hBD2 (0.76 g/l) with the highest percentage of fusion protein in soluble proteins (62.2%) was obtained in the present work, which was the highest yield of hBD2 reported so far.
Zhong
Zhixia
Z
Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou 310027, PR China.
Xu
Zhinan
Z
Peng
Li
L
Huang
Lei
L
Fang
Xiangming
X
Cen
Peilin
P
D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D020080:Tandem Repeat Sequences / Q000235:genetics*; D013696:Temperature; D023083:beta-Defensins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; C106641:DEFB4A protein, human; D011993:Recombinant Fusion Proteins; D023083:beta-Defensins