Bioactive metabolites produced by the Burkholderia genus, especially within the Burkholderia pseudomallei-thailandensis-mallei (Bptm) group, are vital for survival in diverse environments, influencing ecological interactions and contributing to the adaptability of these bacteria. This study focused on the screening of bacteria for antibacterial activity, further culturing in broth. The cell free extract was treated with the equal amount of ethyl acetate where after Vortex the organic phase was vacuum dried and subjected to for the functional group analysis via FTIR Where it revealed the presence of peak at 3352 cm-1, 2177 cm-1 and 1637 cm-1 corresponding to amine, nitrile alkene group respectively. GC-MS analysis presence of 24 bioactive compounds, followed by the ADME and toxicity test were done to narrow down the metabolites. Finally, the docking study was done against Glutathione S-Transferase. Glutathione S-transferase (GST) as the target enzyme because GSTs are key detoxification proteins that conjugate glutathione to xenobiotics and reactive intermediates, thereby protecting bacteria from oxidative damage and chemical stress. In E. coli, GST activity has been implicated in tolerance to multiple antibiotics and other toxic compounds by detoxifying or neutralising them, which can indirectly contribute to antimicrobial resistance. Targeting GST therefore provides a mechanistically relevant way to evaluate whether microbes-derived metabolites can interfere with this detoxification system and potentially sensitise bacteria to existing antibiotics. Four bioactive compounds were docked against GST and Cyclobutyl tridecyl phthalate exhibited the highest score of -8.9 kcal/mol followed by Phthalic acid, 4-cyanophenyl nonyl ester -7 kcal/mol, 1,3-Dioxolane, 4-ethyl-5-octyl-2,2-bis(trifluoromethyl)-, trans- with -6.9 kcal/mol, Pentalamide with a score of -6.3 kcal/mol and Meta-chlorambucil was used as control. The in-vivo evaluation can be performed to evaluate the present study.