Background: The prevalence of multi-drug-resistant E. coli isolates is on the increase around the world. This study was designed to evaluate the antimicrobial activity of Tetrapleura tetraptera (Uyayak) fruit bioactive compounds against E. coli using an in silico approach. Methods: Gas chromatography coupled to mass spectrophotometry (GC-MS) was used to identify the bioactive compounds in our sample T. tetraptera. The resulting bioactive compounds from GC-MS were converted into canonical strings and used for target and absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties predictions using the pkCSM and SWISSADME tools, respectively. Bioactive compounds that met Lipinski's rule of five (ROF) were docked against the dihydropteroate synthase of E. coli using the AutoDock Vina tool, and the resulting interactions were visualized in 2-D using Biovia Discovery Studio 21. Results: The GC-MS analysis returned a total of twenty-eight (28) bioactive compounds, out of which 13 did not violate Lipinski's ROF. ADMET analysis of the screened bioactive compounds showed better absorption (intestinal and water solubility) and toxicity (AMES and hepatoxicity) profiles than trimethoprim, the control drug. Molecular docking gave docking scores that ranged from −4.0 to −5.3 kcal/mol for the bioactive compounds while that of trimethoprim was −6.5 kcal/mol. Target prediction showed that all the bioactive compounds are capable of reaching various targets, with the nuclear receptor being the most abundant target. Conclusion: The bioactive compounds of T. tetraptera fruits examined showed favorable docking scores and pharmacokinetics, suggesting the need for further studies to validate their potential as antimetabolites.