To further validate our methodology, we tested the Best(CCR+DHFR) parameters on the reference calculations and the results obtained using BestCCR parameters, with only minor deviations in the product region. This cross-system validation demonstrates the robustness of our optimization approach and suggests that parameters developed for one hydride transfer system can perform well in mechanistically similar reactions. Thus, the PMF profiles for the hydride transfer reactions in both enzymatic systems demonstrate the success of our optimization strategy.