Cloning and expression of a novel insulin-releasing peptide, brevinin-2GU from Escherichia coli

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• 作者：Qing-feng  ; Zhou¹ ; Ming-yue  ; Li² ; Cheng-wei  ; Li¹ ;   ; ^{xi_tao18@hotmail.com}
• 关键词：Brevinin-2GU ; Escherichia coli ; Expression ; Frog skin ; Insulin secretion
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：2009
• 出版时间：April 2009
• 年：2009
• 卷：107
• 期：4
• 页码：460-463
• 全文大小：464 K

文摘

Brevinin-2GU, a member of a brevinin-2 family of antimicrobial peptides isolated from an extract of the skin of the Asian frog, Hylarana guntheri, displays significant insulin-releasing activity. In this study, in order to obtain relatively large quantity of functional brevinin-2GU protein, the coding sequence of brevinin-2GU gene was cloned into pET32a (+) vector and expression as a Trx fusion protein in Escherichia coli. The results indicated that the expression level of the fusion protein could reach up to 45 % of the total cell proteins. the soluble fusion protein collected from the supernatant of the cell lysate was then separated by Ni²⁺-chelating chromatography and cleaved by Factor Xa protease to release mature brevinin-2GU. Our results showed that the final yield of the brevinin-2GU was 1.7?mg, the purified peptide displayed insulin-releasing activity similar to that of the purified brevinin that was reported earlier. This method allows production of sufficiently large amounts of brevinin-2GU peptide for functional and structural characterizations.