Abstract
A beta1,3-galactosyltransferase (WbgO) was identified in Escherichia coli O55:H7. Its function was confirmed by radioactive activity assay and structure analysis of the disaccharide synthesized with the recombinant enzyme. WbgO requires a divalent metal ion, either Mn(2+) or Mg(2+), for its activity and is active between pH 6.0-8.0 with a pH optimum of 7.0. N-acetylglucosamine (GlcNAc) and oligosaccharides with GlcNAc at the non-reducing end were shown to be its preferred substrates and it can be used for the synthesis of type 1 glycan chains from these substrates. Together with a recombinant bacterial GlcNAc-transferase, benzyl beta-lacto-N-tetraoside was synthesized with the purified WbgO to demonstrate the synthetic utility of WbgO.
MeSH terms
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Acetylglucosamine / metabolism*
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Amino Acid Sequence
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Galactosyltransferases / analysis*
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Galactosyltransferases / genetics
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Galactosyltransferases / isolation & purification
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Galactosyltransferases / metabolism*
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Magnesium / metabolism
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Manganese / metabolism
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Molecular Sequence Data
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N-Acetylglucosaminyltransferases / genetics
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N-Acetylglucosaminyltransferases / metabolism
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Oligosaccharides / chemical synthesis
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Oligosaccharides / chemistry
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Oligosaccharides / metabolism*
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Recombinant Proteins / analysis
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Alignment
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Substrate Specificity
Substances
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Bacterial Proteins
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Oligosaccharides
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Recombinant Proteins
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Manganese
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lacto-N-neotetraose
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WbgO protein, E coli
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Galactosyltransferases
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N-Acetylglucosaminyltransferases
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Magnesium
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Acetylglucosamine