Characterization and synthetic application of a novel beta1,3-galactosyltransferase from Escherichia coli O55:H7.
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.
Liu
Xian-wei
XW
National Glycoengineering Research Center and The State Key Laboratory of Microbial Technology, Shandong University, LuNeng Keji Dasha B 604, Shanda Nanlu 29-1, Jinan, Shandong 250100, China.
Xia
Chengfeng
C
Li
Lei
L
Guan
Wan-yi
WY
Pettit
Nicholas
N
Zhang
Hou-cheng
HC
Chen
Min
M
Wang
Peng George
PG
D000117:Acetylglucosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005700:Galactosyltransferases / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity
D001426:Bacterial Proteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D008345:Manganese; C013084:lacto-N-neotetraose; C544093:WbgO protein, E coli; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases; D008274:Magnesium; D000117:Acetylglucosamine