I'm currently working on a program which will determine the activity difference between an original enzyme and a variant with just 1 or 2 variants. Of course, I'm not talking about the "kinetic activity", but some indications of a probable activity difference should be given. The system has to be sensitive enough to determine the probable activity difference between unthought-of variants.

From the information that I've got from my research, there is two option. I can use one of 3 quantitive enzyme reaction analysis tools (such as EVB, FEB and LRA) or their hybrids.

EVB => Empirical Valence Bond

FEB => Free Energy Perturbation

LRA => Linear Response Approximation

Source: https://www.researchgate.net/post/Can-the-activity-of-proteins-be-studied-using-molecular-dynamic-simulations

Now, my questions are: - Does these application be used for such purpose? - For an accurate prediction of enzyme activity difference, are these programs good enough or require developement? - Which one of them can give the best result? - And lastly, are QM/MM methods a must for an accurate answer?

I'm a high school student so sorry if my questions sound dumb. If you can answer my inquiry's I would appreciate it:)

Thanks