Structural Analysis of Laminarin-Derived Oligosaccharides Produced by Transglycosylation of Exo-β-1,3-glucanase ScEXG1 from Saccharomyces cerevisiae
Szu-Yu Kuo, Chih-Chieh Lin, Hsin-Jo Chang, Reuben Wang, Pei-Yin Lin, Ting-Jang Lu, Yi-Chen Lo
TL;DR
This study shows how an enzyme from yeast can create prebiotic sugars from laminarin, revealing new possibilities for producing health-benefiting compounds.
Contribution
The paper identifies new transglycosylation products and their structures using ScEXG1 under mild conditions.
Findings
ScEXG1 produces a trisaccharide DP3 and two tetrasaccharides DP4–1 and DP4–2 from laminaribiose.
Structural analysis reveals DP3 as β-Glc-(1→6)-β-Glc-(1→3)-Glc and DP4 variants.
Transglycosylation occurs efficiently at low enzyme dosage and specific substrate concentrations.
Abstract
Exo-β-1,3-glucanase EXG1 (ScEXG1) from Saccharomyces cerevisiae hydrolyzes β-1,3-glucans and exhibits transglycosylation activity, enabling biocatalytic synthesis of prebiotic oligosaccharides. In this study, recombinant ScEXG1–6 × His was tested with laminaribiose as both the donor and acceptor. Under low-enzyme dosage, transglycosylation products were detected across a range of substrate concentrations (4.38–32.85 mM), with 21.9 mM laminaribiose producing 270.70 ± 10.32 nmol/mL of the main trisaccharide product-DP3 (DP3) after 3 h. Structural elucidation by HPLC-ESI-MS/MS identified DP3 as β-Glc-(1→6)-β-Glc-(1→3)-Glc based on glycosidic and cross-ring fragmentation patterns. Two additional tetrasaccharides were also characterized as β-Glc-(1→3)-β-Glc-(1→6)-β-Glc-(1→3)-Glc (DP4–1) and β-Glc-(1→6)-β-Glc-(1→3)-β-Glc-(1→3)-Glc (DP4–2). The findings highlight ScEXG1’s ability to produce…
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Taxonomy
TopicsEnzyme Production and Characterization · Polysaccharides and Plant Cell Walls · Biofuel production and bioconversion