The crystal structure of the T4 lysozyme L99A/M102Q in complex with N-phenylglycinonitrile was extracted from Protein Data Bank (PDB ID: 2RBN ). Ligand atom parameters were obtained using GAFF2 (version 2.11) and ligand atomic charges were derived using the RESP method with Gaussian 09 or 16. The protein was parameterized with the FF14SB force field solvated in an orthorombic TIP3P water box using tleap with 15 Å distance between the protein and the edge of the box. The simulation protocol included the following steps: 1) 2000 steps of minimization with gradient descent method;