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Single-point energies were calculated on the optimized geometries using the B97X-D hybrid functional 87 and the 6-311 + G(2d,p) basis set with ultrafine integration grids for the QM part and the ff14SB, gaff2 and TIP3P force fields for the MM part (protein, substrate and water, respectively) with electrostatic embedding

The eluent was prepared as follows: Eluent A was prepared by adding 15 mL of 500 mM 1-octanesulfonic acid sodium salt solution and 0.2 mL of trifluoroacetic acid to an appropriate volume of distilled water for HPLC (Nacalai Tesque), made up to a total of 1000 mL