The pSUFER can rapidly scan for suboptimal residues that affect protein folding and these sites may be relevant to enzyme stability and activity since active sites involved in metastable effects or co

The pSUFER algorithm presents a swift approach to detecting suboptimal residues that impede protein folding. Such residues are of great significance to enzyme stability and activity, as the active sites responsible for metastable effects and conformational changes are the driving force behind the enhanced activity of these proteins, rather than their stability. Previous studies have demonstrated the efficacy of the pSUFER algorithm in improving protein design and concentrating FuncLib design computations on the suboptimal regions of underperforming enzymes, thereby augmenting their catalytic efficiency. In this study, which aims to expedite the process of enhancing enzyme activity, we recommend that FuncLib design not only focus on residues related to the enzyme binding pocket, but also on residues identified by pSUFER that are situated at a distance from the ligand. This is particularly important for suboptimal residues located in the loop region of the binding pocket, which are more flexible and may have a significant impact on the overall protein stability, as well as the entry and exit of small molecules into and out of the binding pocket