title: Biochemical characterization of the novel α-1, 3-galactosyltransferase WclR from Escherichia coli O3.
abstract: Glycosyltransferases (GTs) catalyze the formation of regio- and stereo-specific glycosidic linkages between specific sugar donors and recipients. In this study, the function of the gene wclR from the Escherichia coli O3 O-antigen gene cluster that encodes an α 1, 3-galactosyltransferase (GalT) that acts on the linkage Gal α 1, 3-GlcNAc was biochemically characterized. WclR was expressed in E. coli BL21 (DE3), and the enzymatic product was identified by liquid chromatography-mass spectrometry (LC-MS), collision-induced dissociation electrospray ionization ion trap multiple tandem MS (CID-ESI-IT-MS(n)) and galactosidase digestion, using UDP-Gal as the donor substrate and the synthetic acceptor substrate GlcNAc-PP-De (decyl diphosphate N-acetylglucosamine). The physiochemical properties and the substrate specificity of WclR were investigated. WclR is the first bacterial GalT characterized that acts on the linkage Gal α 1, 3-GlcNAc. This study enhanced our knowledge of the diversified functions of GTs and provided a novel enzyme source for possible pharmaceutical application.
authors.affiliation: TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.affiliation: TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Natalia
authors.initials: N
authors.lastname: Utkina
authors.affiliation: TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Zhou
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Leonid
authors.initials: L
authors.lastname: Danilov
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Torgov
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Veselovsky
authors.affiliation: TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, China. Electronic address: fenglu63@nankai.edu.cn.
authors.forename: Lu
authors.initials: L
authors.lastname: Feng
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology*; D005700:Galactosyltransferases / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase
keywords: Escherichia coli; Galactosyltransferase; Mass spectrometry; WclR