文献类型：期刊文献

英文题名：A Novel Method for Efficient Preparation of Mucosal Adjuvant Escherichia coli Heat-Labile Enterotoxin Mutant (LTm) by Artificially Assisted Self-Assembly In Vitro

作者：Liu, Di[1,2];Zhang, Na[1,2];Zheng, Wenyun[3];Guo, Hua[1,2];Wang, Xiaoli[1,2];Wang, Tianwen[1,2];Wang, Ping[1,2];Ma, Xingyuan[1,2]

机构：[1]E China Univ Sci & Technol, Sch Biotechnol, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]E China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China

年份：2016

卷号：179

期号：1

起止页码：33

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20160801980072);WOS:【SCI-EXPANDED(收录号:WOS:000377464900003)】；

基金：This study was supported by the National Natural Science Foundation (30873190, 31300660), Science and Technology Innovation Action Plan of Shanghai (14431904300), Shanghai Pujiang Program (13PJD012), and a foundation for young teachers from Education Ministry of China (20120074120027) and partially supported by the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Heat-labile enterotoxin (LT); Mucosal adjuvant; In vitro preparation; Artificially assisted self-assembly

摘要：As well-known powerful mucosal adjuvant proteins, Escherichia coli heat-labile enterotoxin (LT) and its non-toxic or low-toxic mutants (LTm) are capable of promoting strong mucosal immune responses to co-administered antigens in various types of vaccines. However, due to the complex composition and special structure, the yield of LTm directly from the recombinant genetic engineering strains is quite low. Here, we put forward a novel method to prepare LTm protein which designed, expressed, and purified three kinds of component subunits respectively and assembled them into a hexamer structure in vitro by two combination modes. In addition, by simulated in vivo environment of polymer protein assembly, the factors of the protein solution system which include environment temperature, pH, ionic strength of the solution, and ratio between each subunit were taken into consideration. Finally, we confirmed the optimal conditions of two assembly strategies and prepared the hexamer holotoxin in vitro. These results are not only an important significance in promoting large-scale preparation of the mucosal adjuvant LTm but also an enlightening to produce other multi-subunit proteins.