Fusion Expression of Human Trypsin-1 with DsbA in E. coli
ZHANG Wen-Yong1, WANG Da-Mei1，2, GAN Yi-Ru1, HUANG He1
1Key Laboratory of System Bioengineering of Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; 2Gan & Lee Pharmaceutical Co. Ltd., Beijing 100026, China
Abstract The trypsin obtained from animal sources may contain infectious agents, currently it is in urgent need of production of recombinant trypsin. According to the codon usage frequency table, the bases of human trypsin-1 were changed by degenerate codon, and then the sequences were fused with the DsbA fragment C-terminal in the vector pET-39b(+). The cloned E. coli with recombinant vector achieved expression. Recombinant proteins mainly existed in the form of inclusion bodies, accounting for about 40% of the total bacteria proteins. By washing the pellet proteins, denaturation, renaturation and thrombin activation, we got 1.6 mg activated trypsin per liter fermented liquid with the commercial porcine trypsin as a standard, and achieved efficient human trypsin expression in prokaryotes.
ZHANG Wen-Yong, WANG Da-Mei, GAN Yi-Ru, HUANG He.Fusion Expression of Human Trypsin-1 with DsbA in E. coli[J] Chemcial Industry and Engineering, 2014,V31(6): 75-79
Copyright 2010 by Chemcial Industry and Engineering
2014, Vol. 31 
