Fig 7. Mutagenesis suggests D474 is involved in catalysis and other charged residues crucial for dPNAG binding.

(A) Reducing sugar assay with 2 mg/ml dPNAG purified from S. aureus with 2 μM PgaB constructs over 24 h. Error bars, S.E. from two independent experiments performed in duplicate. (B) E. coli biofilm disruption assay. Error bars represent the S.E. with n = 3. (C) EC₅₀ values from E. coli biofilm disruption assay. ND: EC₅₀ not determined as > 50% biomass remained or no plateau was reached after treatment with 5 μM enzyme for 2 h. Error bars show 95% confidence interval, and the same color scheme in A is used. (D) Transparent surface representation of the Bb-GH binding groove with mutated amino acids shown in stick representation. Note that D364 is fully buried and only visible due to the transparent representation.