Department of Applied Biology, East China University of Science and Technology, Shanghai 200237, China.
Appl Biochem Biotechnol. 2012 Feb;166(3):612-9. doi: 10.1007/s12010-011-9452-3. Epub 2011 Nov 19.
Tandem repeat multimers of Momordica charantia (MC) peptide MC6 were designed and the recombinant plasmid containing 10 copies of MC6 gene was constructed to improve the expression level of MC6 in Escherichia coli. Under the selected conditions of cultivation and induction, the expression level of recombinant TrxA-MC6(10) protein was above 25% of total bacteria protein. This fusion protein was purified and cleaved with HCl (13%, w/v). Either the un-cleaved or cleaved recombinant proteins was analyzed pharmacological activity by alloxan-induced diabetic mice and only the cleaved products of the recombinant protein showed significant hypoglycemic effects. The study provides a convenient and economical method for the large-scale production of anti-diabetic medicines for pharmaceutical applications.
苦瓜(Momordica charantia)六肽(MC6)串联重复多聚体被设计出来,构建了含有 10 个 MC6 基因拷贝的重组质粒,以提高 MC6 在大肠杆菌中的表达水平。在选择的培养和诱导条件下,重组 TrxA-MC6(10)蛋白的表达水平超过了总细菌蛋白的 25%。该融合蛋白被纯化并用 HCl(13%,w/v)切割。未经切割或切割的重组蛋白均通过链脲佐菌素诱导的糖尿病小鼠分析其药理活性,只有重组蛋白的切割产物显示出显著的降血糖作用。该研究为用于药物应用的抗糖尿病药物的大规模生产提供了一种方便和经济的方法。