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Updated: May 26, 2026

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota
Published on: May 23, 2025
Crystal structure, thermostability and temperature-dependent enzymatic activity of an exo-β-d-glucosaminidase from
Katarzyna Biniek-Antosiak1, Daniel Baranowski1, Joanna Śliwiak1
1Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12-14, 61-704 Poznań, Poland.
Abstract:
Pch-GlmA is a hyperthermophilic GH35 exo-β-d-glucosaminidase whose structure closely resembles its archaeal homologs, yet its functional behavior differs markedly. Calorimetric and fluorimetric temperature scans consistently reveal a complex thermodynamic profile of the enzyme, characterized by distinct thermal transitions. The freshly purified protein appears to be monomeric and required thermal annealing to attain its biologically relevant dimeric state. Catalytic activity is observed only above 75 °C, where the enzyme specifically hydrolyses the glycosidic bond of GlcN-GlcNAc. These findings support a sequential role for Pch-GlmA alongside Pch-Dac in the processing of chitin-derived carbohydrates. Comparison with related GlmA proteins demonstrates that substantial structural similarity does not necessarily translate into equivalent enzymatic properties and that hyperthermophilic enzymes may operate within narrow temperature ranges. Overall, this work underscores the importance of experimental validation when interpreting or predicting the activity of enzymes derived from extremophiles.
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