LARGE-SCALE PRODUCTION OF RECOMBINANT ALPHA-1,2-MANNOSYLTRANSFERASE FROM ESCHERICHIA-COLI FOR THE STUDY OF ACCEPTOR SPECIFICITY AND USE OF THE RECOMBINANT WHOLE CELLS IN SYNTHESIS

We report the first large scale heterologous expression of a recombina nt yeast alpha-1,2-mannosyltransferase in E. coli. The enzyme was isol ated from 10-L and 50-L fermentations, purified and used for mannosyla tion reactions. The specificity of the recombinant enzyme was extensiv ely studied by using mannose derivatives, oligosaccharides, and analog s as accepters, and the results show that the enzyme exhibits high act ivities toward ManOMe and disaccharides connected by an alpha-1,2-mann osidic linkage. The recombinant E. coli cells were also used as a cata lyst for glycosylation reaction, and mannosylation of saccharides and glycopeptides proceeded in moderate to good yields.