Identification and biochemical characterization of a novel α-1,3-mannosyltransferase WfcD from Escherichia coli O141

Chao Chen; Xi Hou; Natalia Utkina; Leonid Danilov; Dawei Zhou; Vladimir Torgov; Vladimir Veselovsky; Bin Liu; Lu Feng

doi:10.1016/j.carres.2017.04.003

Identification and biochemical characterization of a novel α-1,3-mannosyltransferase WfcD from Escherichia coli O141

Carbohydr Res. 2017 Apr 18:443-444:78-86. doi: 10.1016/j.carres.2017.04.003. Epub 2017 Apr 6.

Authors

Chao Chen¹, Xi Hou¹, Natalia Utkina², Leonid Danilov², Dawei Zhou¹, Vladimir Torgov², Vladimir Veselovsky², Bin Liu¹, Lu Feng³

Affiliations

¹ 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.
² N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation.
³ 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.

PMID: 28402841
DOI: 10.1016/j.carres.2017.04.003

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.

Keywords: Escherichia coli; Mannosyltransferase; Mass spectrometry; WfcD.

MeSH terms

Biocatalysis
Carbohydrate Sequence
Detergents / pharmacology
Escherichia coli / enzymology*
Kinetics
Mannosyltransferases / metabolism*
Metals / pharmacology
O Antigens / chemistry
O Antigens / metabolism
Substrate Specificity

Substances

Detergents
Metals
O Antigens
Mannosyltransferases