Yanhong Li1, Mengyang Xue2, Xue Sheng1, Hai Yu1, Jie Zeng3, Vireak Thon4, Yi Chen1, Musleh M Muthana1, Peng G Wang5, Xi Chen6. 1. Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA. 2. 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. 3. 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. 4. Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA; Department of Chemistry, Georgia State University, Atlanta, GA 30303, USA. 5. 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. 6. Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA. Electronic address: xiichen@ucdavis.edu.
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.
β1-3-N-Acetylglucosaminyltransferases (β3n class="Chemical">GlcNAcTs) 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: Zhigang Wu; Yunpeng Liu; Cheng Ma; Lei Li; Jing Bai; Lauren Byrd-Leotis; Yi Lasanajak; Yuxi Guo; Liuqing Wen; He Zhu; Jing Song; Yanhong Li; David A Steinhauer; David F Smith; Baohua Zhao; Xi Chen; Wanyi Guan; Peng George Wang Journal: Org Biomol Chem Date: 2016-11-29 Impact factor: 3.876
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