Biochemical characterization of the novel α-1, 3-galactosyltransferase WclR from Escherichia coli O3

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

doi:10.1016/j.carres.2016.04.012

Biochemical characterization of the novel α-1, 3-galactosyltransferase WclR from Escherichia coli O3

Carbohydr Res. 2016 Jul 22:430:36-43. doi: 10.1016/j.carres.2016.04.012. Epub 2016 Apr 13.

Authors

Chao Chen¹, Bin Liu², Yongchang Xu¹, Natalia Utkina³, Dawei Zhou¹, Leonid Danilov³, Vladimir Torgov³, Vladimir Veselovsky³, Lu Feng⁴

Affiliations

¹ TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China.
² TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, China.
³ N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia.
⁴ 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.

PMID: 27196310
DOI: 10.1016/j.carres.2016.04.012

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.

Keywords: Escherichia coli; Galactosyltransferase; Mass spectrometry; WclR.

MeSH terms

Amino Acid Sequence
Escherichia coli / enzymology*
Galactosyltransferases / chemistry*
Galactosyltransferases / metabolism*
Hydrogen-Ion Concentration
Stereoisomerism
Substrate Specificity

Substances

Galactosyltransferases
N-acetyllactosaminide alpha-1,3-galactosyltransferase