Extended Data Fig. 8: Analysis of the activity and stability of BT31776S-GlcNAc and mutant variants.
From: Sulfated glycan recognition by carbohydrate sulfatases of the human gut microbiota
a, High pressure anion exchange chromatography (HPAEC) of wild-type BT31776S-GlcNAc wild-type (WT) and substituted variants. The produced product is highlighted by a grey box. b, Thin layer chromatography (TLC) analysis of WT BT31776S-GlcNAc and its mutants. Asterisks are placed above lanes where activity is observed. Both HPAEC (a) and TLC (b) reactions utilised 6 mM substrate and 5 μM enzyme, with 3 mM HEPES, 45 mM NaCl, and 5 mM CaCl2 over a 48 h period at 37 °C. Control represents the substrate incubated in same conditions without enzyme. c, DSF analysis showing relative thermostability of mutant proteins of BT31776S-GlcNAc in comparison to the WT enzyme. Experiments are technical triplicates and error bars represent SEM.
