title: Donor substrate promiscuity of bacterial β1-3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1-4-galactosyltransferases.
abstract: β1-3-N-Acetylglucosaminyltransferases (β3GlcNAcTs) and β1-4-galactosyltransferases (β4GalTs) have been broadly used in enzymatic synthesis of N-acetyllactosamine (LacNAc)-containing oligosaccharides and glycoconjugates including poly-LacNAc, and lacto-N-neotetraose (LNnT) found in the milk of human and other mammals. In order to explore oligosaccharides and derivatives that can be synthesized by the combination of β3GlcNAcTs and β4GalTs, donor substrate specificity studies of two bacterial β3GlcNAcTs from Helicobacter pylori (Hpβ3GlcNAcT) and Neisseria meningitidis (NmLgtA), respectively, using a library of 39 sugar nucleotides were carried out. The two β3GlcNAcTs have complementary donor substrate promiscuity and 13 different trisaccharides were produced. They were used to investigate the acceptor substrate specificities of three β4GalTs from Neisseria meningitidis (NmLgtB), Helicobacter pylori (Hpβ4GalT), and bovine (Bβ4GalT), respectively. Ten of the 13 trisaccharides were shown to be tolerable acceptors for at least one of these β4GalTs. The application of NmLgtA in one-pot multienzyme (OPME) synthesis of two trisaccharides including GalNAcβ1-3Galβ1-4GlcβProN3 and Galβ1-3Galβ1-4Glc was demonstrated. The study provides important information for using these glycosyltransferases as powerful catalysts in enzymatic and chemoenzymatic syntheses of oligosaccharides and derivatives which can be useful probes and reagents.
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA; National Glycoengineering Research Center and Shandong Province Key Laboratory of Carbohydrate Chemistry and Glycobiology, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Mengyang
authors.initials: M
authors.lastname: Xue
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Xue
authors.initials: X
authors.lastname: Sheng
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Hai
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA; School of Food Science, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zeng
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA; Department of Chemistry, Georgia State University, Atlanta, GA 30303, USA.
authors.forename: Vireak
authors.initials: V
authors.lastname: Thon
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA.
authors.forename: Musleh M
authors.initials: MM
authors.lastname: Muthana
authors.affiliation: National Glycoengineering Research Center and Shandong Province Key Laboratory of Carbohydrate Chemistry and Glycobiology, Shandong University, Jinan, Shandong 250100, China; Department of Chemistry, Georgia State University, Atlanta, GA 30303, USA.
authors.forename: Peng G
authors.initials: PG
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA. Electronic address: xiichen@ucdavis.edu.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
mesh_terms: D002236:Carbohydrate Conformation; D005700:Galactosyltransferases / Q000378:metabolism*; D016480:Helicobacter pylori / Q000201:enzymology*; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000201:enzymology*; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry; D013379:Substrate Specificity
chemical_list: D009844:Oligosaccharides; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases
keywords: Acceptor substrate specificity; Carbohydrate; Donor substrate promiscuity; Enzymatic synthesis; Glycosyltransferase; One-pot multienzyme