## 3 Results
### In Silico Prediction of Mechanism by Molecular Docking Analysis  
Ligand-based molecular docking is a computational technique that can be used, among other methods, as a starting point for understanding the targeted mechanism of action of a given molecular structure. In the form of free-binding energy values, the obtained results may indicate an increased/decreased affinity of the analyzed molecule towards the selected target compared to the native ligand (a known inhibitor), given that the binding energy decreases when the compounds' affinity increases [64, 65, 66]. For our current study, we used a molecular docking-based protocol to identify possible protein targets for the 37 RGEO components, whose inhibition could be correlated with their in vitro antimicrobial \begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Bacterial and Yeast** & **Disk Diffusion (mm)** & **MIC Value (mg/mL)** & **MBC Value (mg/mL)** & **MFC Value (mg/mL)** \\ \hline _Streptococcus mutans_ & 19.77 \(\pm\) 0.26 b  & 1.25 & 1.25 & - \\ \hline _Streptococcus pyogenes_ & 20.61 \(\pm\) 0.21 a  & 1.25 & 1.25 & - \\ \hline _Staphylococcus aureus_ & 19.89 \(\pm\)0.14 b  & 2.5 & 2.5 & - \\ \hline _Enterococcus faecalis_ & 16.56 \(\pm\) 0.17 d  & 2.5 & 2.5 & - \\ \hline _Escherichia coli_ & 15.76 \(\pm\) 0.1 fe  & 5.0 & 5.0 & - \\ \hline _Klebsiella pneumoniae_ & 15.71 \(\pm\) 0.13 fe  & 5.0 & 5.0 & - \\ \hline _Salmonella enterica_ & 15.65 \(\pm\) 0.23 fe  & 5.0 & 5.0 & - \\ \hline _Shigella flexneri_ & 15.21 \(\pm\) 0.14 f  & 5.0 & 5.0 & - \\ \hline _Pseudomonas aeruginosa_ & 15.95 \(\pm\) 0.08 de  & 5.0 & 5.0 & - \\ \hline _Candidia albicans_ & 18.66 \(\pm\) 0.26 c  & 2.5 & - & 2.5 \\ \hline _Candidia parapsilosis_ & 18.42 \(\pm\) 0.39 c  & 2.5 & - & 2.5 \\ \hline \hline \end{tabular}  
Values with different superscript are significantly different (\(p<\) 0.05) according to Tukey test; each value is the Mean \(\pm\) SD.  
\end{table} 
Table 6: Antimicrobial of the RGEO by disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC). activity. The same method was also employed to provide an insight regarding a potential in vitro protein targeted based antioxidant activity, apart from the chemical structure related antioxidant activity described above. Protein targets, usually associated with bactericidal/bacteriostatic effects, such as dihydropyroate synthase (DHPS), dihydrofolate reductase (DHFR), D-alanine: D-alanine ligase (Ddl), penicillin binding protein 1a (PBP1a), DNA gyrase, type IV topoisomerase, and isoleucyl-tRNA synthetase (IARS), were used in the present work [10]. Furthermore, molecular docking screening was also employed to assess the RGEO components inhibitory potential towards protein targets that play active roles in intracellular antioxidant mechanisms. To achieve this goal CYP2C9, lipoxygenase xanthine oxidase, and NADPH-oxidase [67] were selected as protein targets. Docking scores recorded as \(\Delta\)G values (kcal/mol) corresponding to the 37 docked compounds and the native ligands of each protein are presented in Table 7. However, we intended to spot a trend related to a possible protein-targeted cumulative mechanism of action associated with the set of 37 RGEO compounds in the sense that if a majority of the RGEO components score better or comparable docking results with the native ligand of a target protein, then that cumulative effect may lead to the observed biological activity. To better visualize this tendency, firstly, the docking scores corresponding to each protein target column were reordered in descending order, the lowest \(\Delta\)G value representing the highest affinity for that specific target. Subsequently, we generated a heatmap based on the rearranged table.