Faisal Nureldin Awad1, Pedro Laborda2, Meng Wang2, Ai Min Lu3, Qian Li2, Zhi Peng Cai2, Li Liu4, Josef Voglmeir5. 1. Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, PR China; National Food Research Centre, Khartoum North, P.O. Box 213, Sudan. 2. Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, PR China. 3. College of Sciences, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, PR China. 4. Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, PR China; Qlyco Ltd., 50 Zhonglingjie, Nanjing 210014, PR China. Electronic address: liu.lichen@qlyco.com. 5. Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, PR China. Electronic address: josef.voglmeir@njau.edu.cn.
Abstract
BACKGROUND: Mannoside phosphorylases are frequently found in bacteria and play an important role in carbohydrate processing. These enzymes catalyze the reversible conversion of β-1,2- or β-1,4-mannosides to mannose and mannose-1-phosphate in the presence of inorganic phosphate. METHODS: The biochemical parameters of this recombinantly expressed novel mannose phosphorylase were obtained. Furthermore purified reaction products were subjected to ESI- and MALDI-TOF mass spectrometry and detailed NMR analysis to verify this novel type of β-1,3-mannose linkage. RESULTS: We describe the first example of a phosphorylase specifically targeting β-1,3-mannoside linkages. In addition to mannose, this phosphorylase originating from the bacterium Zobellia galactanivorans could add β-1,3-linked mannose to various other monosaccharides and anomerically modified 5-bromo-4-chloro-3-indolyl-glycosides (X-sugars). CONCLUSIONS: An unique bacterial phosphorylase specifically targeting β-1,3-mannoside linkages was discovered. GENERAL SIGNIFICANCE: Functional extension of glycoside hydrolase family 130.
BACKGROUND: Mannoside phosphorylases are frequently found in bacteria and play an important role in carbohydrate processing. These enzymes catalyze the reversible conversion of β-1,2- or β-1,4-mannosides to mannose and mannose-1-phosphate in the presence of inorganic phosphate. METHODS: The biochemical parameters of this recombinantly expressed novel mannose phosphorylase were obtained. Furthermore purified reaction products were subjected to ESI- and MALDI-TOF mass spectrometry and detailed NMR analysis to verify this novel type of β-1,3-mannose linkage. RESULTS: We describe the first example of a phosphorylase specifically targeting β-1,3-mannoside linkages. In addition to mannose, this phosphorylase originating from the bacterium Zobellia galactanivorans could add β-1,3-linked mannose to various other monosaccharides and anomerically modified 5-bromo-4-chloro-3-indolyl-glycosides (X-sugars). CONCLUSIONS: An unique bacterial phosphorylase specifically targeting β-1,3-mannoside linkages was discovered. GENERAL SIGNIFICANCE: Functional extension of glycoside hydrolase family 130.
Authors: Ao Li; Elisabeth Laville; Laurence Tarquis; Vincent Lombard; David Ropartz; Nicolas Terrapon; Bernard Henrissat; David Guieysse; Jeremy Esque; Julien Durand; Diego P Morgavi; Gabrielle Potocki-Veronese Journal: Microb Genom Date: 2020-10