To perform the classical molecular dynamics for PfK13, the initial coordinates were obtained from the reported crystal structure (PDB ID:4yy8). For the mutant structures, all the four mutants were generated using the mutagenesis function of Pymol by substituting the residue with the mutant residue. Simulations were performed on the monomeric state of PfK13, using the NAMD simulation package. AMBER FF14SB force-field was used to generate the force fields of WT and mutant structures. The systems were immersed in the cubic box of TIP3P water molecules maintaining at least 15Å of the separation between the solute and the edges of the box. The system was centred in the periodic box of dimensions of 90.53 x 118.56 x 84.13 Å 3 containing 23190 water molecules. The 10N a + and 6Cl -counter ions were added to the water to make the overall charge neutrality of the system. The Particle Mesh Ewald (PME) approach was employed for the long-range interactions and a cut-off value of 10Å were used for the short-range interactions. The system was minimized at 0K for 0.1ns keeping the heavy atoms fixed. Further, the system was equilibrated at a constant temperature, i.e., at 300K and 1 atm pressure for 0.25ns. The simulation was carried out under NPT ensembles for 400ns. The initial 200ns were performed as equilibration by removing the constraint force and allowing the free movement of the atoms. Subsequently, 200ns were treated as production dynamics for the structural analysis. The resulting trajectories were visualized and analyzed using the VMD software.