%0 Journal Article
%T Soluble Expression and Purification of Human Alpha-Defensin-5 in Escherichia coli<br>人α防御素5在大肠杆菌中的可溶性表达与纯化研究
%A Aiping Wang
%A Yongping Su
%A Tianmin Cheng
%A Zhongmin Zou
%A Junping Wang
%A <br>王艾平
%A 粟永萍
%A 程天民
%A 邹仲敏
%A 王军平
%J 生物工程学报
%D 2008
%I 
%X DNA fragment containing human alpha-defensin 5 mature peptide (mHD-5) coding sequence with biased codons of E. coli was amplified by PCR, which was subsequently cloned into the plasmid pMAL-p2x in order to create pMAL-p2x-mHD-5 expression vector. The plasmid pMAL-p2x-mHD-5 was transferred into engineered strain BL21(DE3) to express heterogeneous fusion protein (MBP-mHD-5). The soluble MBP-mHD-5 targeted protein inducible expressed by IPTG was accounted for about 30% under optimized conditions. The recombinant mHD-5 (rmHD-5) peptide was successfully purified through a separation process including affinity chromatography, Factor Xa digestion and ion exchange chromatography. The bioactivity of rmHD-5 was examined by bacteria-inhibition tests in liquid culture. The growth of E. coli ATCC25922 was dramatically suppressed with an inhibition rate of 90%, with the presence of 62.5mg/mL rmHD-5 in the media. These results indicate that the strategy of soluble expression of fusion protein in E. coli can be a useful and practical way to produce bioactive defensins.
%K defensin
%K prokaryotic expression
%K purify
%K bioactivity<br>防御素
%K 原核表达
%K 纯化
%K 生物活性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=37E6D4AC664F084F001F4DF828DCDFDC&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=0B39A22176CE99FB&sid=6490F0E20C4B41AD&eid=D0182A31A5EB14BA&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=12