We have used five CVs for the controlled sampling approach. Among the CVs used, the center of the mass distance between the stem regions was used as an umbrella sampling window coordinate. Further, in each window, the sampling is accelerated by parallel bias metadynamics by the other four collective variables (subsidiary CVs): radius of gyration, hydrogen bonds, ermsd and contacts. The protocol was carried out for each specific system of amino acids containing D-and L-stereoisomers. A potential harmonic restraint of 200 kJ/mol/nm 2 was applied to the umbrella sampling coordinate to make sure that the value of the reaction coordinate fluctuates around the succussive positions where potential is applied. Windows were constructed from folded state to unfolded state from the umbrella sampling coordinate value range from 0.8 nm (folded state) to 4.6 nm (unfolded state) with a difference of 0.2 nm between the windows. For the parallel bias metadynamics, a bias factor of 10 and an initial hill height of 0.5 kJ/mol were used. Further, the Gaussian widths of 0.1, 0.4, 1, and 0.05 were used respectively for ermsd, hydrogen bonds, contacts, and radius of gyration, respectively. A total of 1 𝜇𝑠 simulation was carried out per system by constructing 20 windows across the umbrella sampling coordinate and by simulating each window for 50 ns. Finally, the free energy surface was constructed by combining the bias contribution from the umbrella sampling potential and the potential from the parallel bias metadynamics from each window using the weighted histogram analysis method (WHAM) . The results are interpreted against the ermsd parameter and stem distance (umbrella sampling coordinate). The ermsd parameter is well suited to explain various interactive folded states, base pairing schemes, and base stacking schemes in the literature .