Synthesis of the milk oligosaccharide 2'-fucosyllactose using recombinant bacterial enzymes.
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.
Albermann
C
C
Institut für chemische Mikrobiologie, Bergische Universität GH, FB9, Gauss-Strasse 20, D-42097, Wuppertal, Germany.
Piepersberg
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W
Wehmeier
U F
UF
D000818:Animals; D004926:Escherichia coli / Q000201:enzymology; D005647:Fucosyltransferases / Q000378:metabolism*; D006154:Guanosine Diphosphate Fucose / Q000096:biosynthesis* / Q000378:metabolism*; D016480:Helicobacter pylori / Q000235:genetics / Q000378:metabolism; D006836:Hydro-Lyases / Q000378:metabolism; D007785:Lactose / Q000378:metabolism; D008892:Milk / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014312:Trisaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
D011994:Recombinant Proteins; D014312:Trisaccharides; D006154:Guanosine Diphosphate Fucose; D005647:Fucosyltransferases; D006836:Hydro-Lyases; C044971:GDPmannose 4,6-dehydratase; D007785:Lactose; C031420:2'-fucosyllactose