title: Identification and biochemical characterization of a novel α-1,3-mannosyltransferase WfcD from Escherichia coli O141.
abstract: Glycosyltransferases (GTs) catalyze the formation of regio- and stereospecific glycosidic linkages between specific sugar donors and recipients. In this study, the function of the wfcD gene from the Escherichia coli O141 O-antigen gene cluster encoding an α-1,3-mannosyltransferase that catalyzed the formation of the linkage Man(α1-3)-GlcNAc was biochemically characterized. WfcD 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-MSn) and glycosidase digestion using the donor substrate GDP-Man and the synthetic acceptor substrate decyl diphosphate 2-acetamido-2-deoxy-α-D-glucopyranose (GlcNAc-PP-De). The kinetic and physiochemical properties and the substrate specificity of WfcD were investigated. WfcD is the first characterized bacterial mannosyltransferase that acts on the Man(α1-3)-GlcNAc linkage. This study enhances our knowledge of the diverse functions of GTs.
authors.affiliation: TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin, 300457, PR China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, 23 Hongda Street, TEDA, Tianjin, 300457, PR China.
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.affiliation: TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin, 300457, PR China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, 23 Hongda Street, TEDA, Tianjin, 300457, PR China.
authors.forename: Xi
authors.initials: X
authors.lastname: Hou
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
authors.forename: Natalia
authors.initials: N
authors.lastname: Utkina
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
authors.forename: Leonid
authors.initials: L
authors.lastname: Danilov
authors.affiliation: TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin, 300457, PR China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, 23 Hongda Street, TEDA, Tianjin, 300457, PR China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Zhou
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Torgov
authors.affiliation: N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Veselovsky
authors.affiliation: TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin, 300457, PR China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, 23 Hongda Street, TEDA, Tianjin, 300457, PR China.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: TEDA Institute of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin, 300457, PR China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, 23 Hongda Street, TEDA, Tianjin, 300457, PR China. Electronic address: fenglu63@nankai.edu.cn.
authors.forename: Lu
authors.initials: L
authors.lastname: Feng
mesh_terms: D055162:Biocatalysis; D002240:Carbohydrate Sequence; D003902:Detergents / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D008364:Mannosyltransferases / Q000378:metabolism*; D008670:Metals / Q000494:pharmacology; D019081:O Antigens / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D003902:Detergents; D008670:Metals; D019081:O Antigens; D008364:Mannosyltransferases
keywords: Escherichia coli; Mannosyltransferase; Mass spectrometry; WfcD