Large scale production of recombinant .alpha.-1,2-mannosyltransferase from E. coli for the study of acceptor specificity and use of the recombinant whole cells in synthesis
作者:Guido F. Herrmann、Peng Wang、Gwo-Jenn Shen、Eduardo Garcia-Junceda、Shaheer H. Khan、Khushi L. Matta、Chi-Huey Wong
DOI:10.1021/jo00100a043
日期:1994.10
We report the first large scale heterologous expression of a recombinant yeast alpha-1,2-mannosyltransferase in E. coli. The enzyme was isolated from 10-L and 50-L fermentations, purified and used for mannosylation reactions. The specificity of the recombinant enzyme was extensively studied by using mannose derivatives, oligosaccharides, and analogs as accepters, and the results show that the enzyme exhibits high activities toward ManOMe and disaccharides connected by an alpha-1,2-mannosidic linkage. The recombinant E. coli cells were also used as a catalyst for glycosylation reaction, and mannosylation of saccharides and glycopeptides proceeded in moderate to good yields.