초록
<P>GDP-L-fucose is an L-fucose donor to synthesize fucosylated compounds such as human milk oligosaccharides or Lewis antigen. In this study, we used Lactococcus lactis subsp. cremoris NZ9000 to express 4 enzymes, ManB, ManC, Gmd, and WcaG and produced GDP-L-fucose by using one-pot synthesis method with mannose-6-phosphate as substrate and the enzymes as biocatalyst. For preparation of enzyme mixture, 4 genes (manB, manC, gmd, and wcaG) cloned from Escherichia coli were transformed into L. lactis strains using pNZ8008 and the recombinant cell lysates were obtained after cultivation. When mannose-6-phosphate was used as the substrate, the consecutive reactions with ManB, ManC, Gmd, and WcaG resulted in the successful production of GDP-L-fucose (0.13 mM). When GDP-D-mannose was used as the substrate, it was entirely converted to GDP-L-fucose (0.2 mM; 0.12 g/L) via 2 enzymatic reactions mediated by Gmd and WcaG. This is the first report of GDP-Lfucose production by using multiple enzymes expressed in lactic acid bacteria.</P>