Simulation analysis was performed using the CPPTRAJ 41 module from AMBER 2020 and the Python library MDAnalysis. Snapshots were collected every 50 ps from the conventional MD production runs for further analysis. Open cleft volume analysis was performed using POcket Volume MEasurer (POVME) 3.0, 44 by considering 1,000 frames over all replicas, taken every 2.5 ns of the production run. Root mean square fluctuations (RMSF) were calculated on the Cα-atoms along the amino acid sequence of each protein relative to their averaged position over time. Hydrogen bonds for catalytic residues were calculated by considering a donor-acceptor distance cut-off of 3.5 Å and a donor-hydrogen-acceptor angle cut-off of 120°, considering all 25,000 frames collected per replica. The same number of frames was employed for calculating the change in distance between the Cγ-atoms of the conserved tryptophan in the W-loop and the conserved tyrosine with which it forms an active site aromatic clamp. Lastly, Shortest Path Map (SPM) analysis was carried out by means of the DynaComm.py Python script, using the dynamic cross correlation matrices and average inter-residue distance matrices for the MD simulations.