Fig. 3: Mutational analysis of relevant residues of Fl8CE20_II.
From: Insights into a water-mediated catalytic triad architecture in CE20 carbohydrate esterases
a Closeup of the active site region, showcasing mutated residues (highlighted in a purple box) and relevant residues that directly interact with them through hydrogen bonds. The coordinated water is represented as a red sphere. b Kinetic parameters of selected residues, represented as bubble chart, where the X- and Y-axis represent KM and kcat, respectively, and the bubbles represent the catalytic efficiency in kcat/KM. These experiments were conducted as three independent technical replicates (n = 3). Mean values ± SD are shown in Supplementary Table S5. Selected mutated regions, as shown in (a), displaying how changing the residue alters important hydrogen bonds: D513N (c), K370M (d), D513E (e), D513Q (f), Y334N (g), R313Q (h), R313E (i) and S434A (j). Mutated residues and changes in H-bond distances are highlighted in a purple box. The tertiary structures are represented as cartoons, side chains as coloured sticks (pink: ancillary domain, light teal: catalytic domain, orange: catalytic residues). Dashed lines indicate possible polar interactions (black), potential sterically hindrance (red) and potential loss of interaction due to increased distance (grey).
