牛乳铁蛋白活性多肽LfcinB在大肠杆菌中的串联表达、纯化及生物活性分析  

Tandem Expression,Purification and Bioactivity Characterization of Bovine Lactoferricin Peptide in Escherichia coli

作　　者：刘珂杭 宗西翠 张慧丹 完颜杨珂 陈玉清[1] Liu Kehang;Zong Xicui;Zhang Huidan;Wanyan Yangke;Chen Yuqing(School of Life Sciences,Nanjing Normal University,Institute of Biochemistry and Biological Products,Nanjing 210023,China;Department of Basic Western Medicine,Nanjing University of Chinese Medicine Hanlin College,Taizhou 225300,China)

机构地区：[1]南京师范大学生命科学学院,生物化学与生物制品研究所,江苏南京210023 [2]南京中医药大学翰林学院,医学院西医基础教研室,江苏泰州225300

出　　处：《南京师大学报（自然科学版）》2020年第2期100-107,共8页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金项目(81573337);江苏省自然科学基金项目(BK20141446).

摘　　要：牛乳铁蛋白(LfcinB)是一种阳离子抗菌肽,具有抗细菌、抗肿瘤等多种生物学活性.本文探讨了利用SUMO(Small Ubiquitin-Related Modifier)融合标签在大肠杆菌系统中通过LfcinB串联表达策略制备重组LfcinB的方法.结果显示,大于90%的SUMO-(LfcinB)n蛋白以可溶形式表达于BL21(DE3)细胞中;SUMO-(LfcinB)2的表达量高于SUMO-LfcinB和SUMO-(LfcinB)3.通过SUMO特异性蛋白酶切除SUMO标签,羟胺释放单体,得到纯化的重组LfcinB,产率约为15 mg/L.生物活性结果表明,重组LfcinB具有一定的抗菌和抗肿瘤活性.本研究为结合SUMO标签与串联表达策略大量制备获得具有生物学功能的重组抗菌肽提供了一种有效的方法.Bovine lactoferricin(LfcinB)is a cationic antimicrobial peptides with activities of anti-bacteria and anti-tumor. Here it was reported that a method of producing tandem multimers of LfcinB using SUMO(small ubiquitin-related modifier)tag in Escherichia coli. Results showed that the expression level of the soluble fusion protein with SUMO tag was more than 90% of total target protein,and the dimer of LfcinB was expressed at a higher level than monomer and trimer when fused with SUMO tag. After using SUMO-specific protease to remove the SUMO tag,hydroxylamine to release monomer,the purified LfcinB was obtained with a yield of approximately 15 mg/L. The biological assays demonstrated that the recombinant LfcinB exhibited a certain antimicrobial and antitumor activity. Altogether,the SUMO fusion system with tandem repeat expression technology provides a potential production method for functional antimicrobial peptides.

关 键 词：Bovine lactoferricin 抗菌肽 串联表达 SUMO 

分 类 号：Q291[生物学—细胞生物学]