Identification and biochemical characterization of a novel α-1, 3-mannosyltransferase WfcD from Escherichia coli O141. (18th April 2017)

• Record Type: Journal Article
• Authors: Chen, Chao; Hou, Xi; Utkina, Natalia; Danilov, Leonid; Zhou, Dawei; Torgov, Vladimir; Veselovsky, Vladimir; Liu, Bin; Feng, Lu
• Is Part Of: Carbohydrate research. Volume 443/444(2017)
• Page Start: 78
• Page End: 86
• Publication Date: 2017-04-18
• Subjects: Escherichia coli -- WfcD -- Mannosyltransferase -- Mass spectrometry; Carbohydrates -- Periodicals; Chemistry, Organic -- Periodicals; Biochemistry -- Periodicals; Carbohydrates -- Periodicals; Chimie organique -- Périodiques; Glucides -- Périodiques; Biochemistry; Carbohydrates; Chemistry, Organic; Periodicals; Electronic journals; 507.78
• DOI: 10.1016/j.carres.2017.04.003
• Languages: English
• ISSNs: 0008-6215

Abstract

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-MS n ) 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. Graphical abstract: Highlights: We characterized the function of the wfcD gene, part of the Escherichia coli O141 O-antigen gene cluster. 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.