Discovery and biochemical characterization of a mannose phosphorylase catalyzing the synthesis of novel β-1,3-mannosides

Unravelling the Specificity of Laminaribiose Phosphorylase from Paenibacillus sp. YM‐1 towards Donor Substrates Glucose/Mannose 1‐Phosphate by Using X‐ray Crystallography and Saturation Transfer Difference NMR Spectroscopy

“…These data are in line with those reported by Awad et al. for the same disaccharide arising from GH130 β‐(1→3)‐mannan phosphorylase‐mediated synthesis …”

“…The use of GPs for β‐(1→3)‐mannosylation has been previously conducted by using a GH130 β‐(1→3)‐mannan mannoside phosphorylase (Zg0232) from Zobellia galactanivorans DSM 12802, which transfers mannose from Man1P to a variety of sugar acceptors, including a noncognate acceptor, Glc . In contrast, our study demonstrates the relaxed specificity of Ps LBP towards the sugar 1‐phosphate donor, Man1P, from which mannose was transferred onto a Glc acceptor, resulting in the production of Man‐(1→3)‐Glc disaccharide 1 .…”

“…Understanding the mechanism of GP action on both natural and noncognate substrates would therefore provide background knowledge that could underpin applications of GPs in carbohydrate synthesis, in both academic and industrial settings. Unlike other conventional substrate screening experiments, which have been conducted by various groups on GPs, we aimed to pinpoint the mechanism by which Ps LBP recognised and utilised Man1P as its noncognate donor, through X‐ray crystallography and STD NMR spectroscopy.…”

Analysis of the diversity of the glycoside hydrolase family 130 in mammal gut microbiomes reveals a novel mannoside-phosphorylase function

“…The newly created GT108 family only contains six characterized members, all acting on mannogen, a linear β-1,2polymannoside. In contrast, the GH130 family is much more polyspecific, with 18 characterized members, of which 3 are β-1,2mannosidases [12,13] and 14 are glycoside phosphorylases acting on a large variety of substrates, such asβ-1,4-mannooligosaccharides [14][15][16][17][18], β-1,4mannosyl-N-acetyl-glucosamine [19],β-1,4mannosyl-N,N'-diacetylchitobiose [20],β-1,2-mannobiose [21,22],β-1,2-oligomannans [21] andβ-1,3-mannobiose [23]. Their large diversity of specificities, the tolerance of some enzymes towards acceptors, and their ability to generate αMan1P through phosphorolysis of cheap substrates for the synthesis of various heteromannosides, makes GH130 glycoside phosphorylases attractive enzymes for the in vitro synthesis of β-mannosidic linkages, which are considered to be some of the most challenging glycosidic linkages in synthetic carbohydrate chemistry [24].…”

“…In order to predict the specificity of enzymes in this family that are still uncharacterized, in 2013 we proposed a classification into two subfamilies, namely GH130_1, strictly acting on β-1,4-mannosyl-glucose, and GH130_2, targeting β-1,4-mannosyl-N,N′-diacetylchitobiose (the core oligosaccharide of N-glycans), or β-1,4-mannooligosaccharides [20]. This subfamily analysis also revealed one heterogeneous GH130_NC sequence cluster, which was later shown to include enzymes of various specificities and mechanisms, such as β-1,2-mannosidases [12, 13], β-1,2-mannobiose- and β-1,2-oligomannan-phosphorylases [21, 22], and a β-1,3-mannobiose-phosphorylase [23]. Even though these early subfamilies are still used [17], we wished to perform a new analysis based on the greater sequence diversity available today.…”