Synthesis of the milk oligosaccharide 2'-fucosyllactose using recombinant bacterial enzymes |
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Authors: | Albermann C Piepersberg W Wehmeier U F |
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Affiliation: | Institut für chemische Mikrobiologie, Bergische Universit?t GH, FB9, Gauss-Strasse 20, D-42097, Wuppertal, Germany. |
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Abstract: | The enzymatic synthesis of GDP-beta-L-fucose and its enzymatic transfer reaction using recombinant enzymes from bacterial sources was examined. The GDP-D-mannose 4,6-dehydratase and the GDP-4-keto-6-deoxy-D-mannose 3,5-epimerase-4-reductase from Escherichia coli K-12, respectively, were used to catalyse the conversion of GDP-alpha-D-mannose to GDP-beta-L-fucose with 78% yield. For the transfer of the L-fucose to an acceptor, we cloned and overproduced the alpha-(1-->2)-fucosyltransferase (FucT2) protein from Helicobacter pylori. We were able to synthesise 2'-fucosyllactose using the overproduced FucT2 enzyme, enzymatically synthesised GDP-L-fucose and lactose. The isolation of 2'-fucosyllactose was accomplished by anion-exchange chromatography and gel filtration to give 65% yield. |
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