The increasing incidence of antimicrobial drug resistance is a major problem in the health sector, and this has encouraged the discovery of novel antibacterial agents present in natural products that can overcome this problem. Since ancient times, propolis has been widely used as a traditional medicine for various diseases, particularly those caused by bacteria. Propolis is a bee product composed of a mixture of resin derived from plant exudates and enzymes derived from bee saliva. Stingless bees produce more propolis than honey and are widely distributed in tropical areas like Indonesia. The aim of this study was to evaluate the antibacterial activity of ethanolic extract propolis (EEP) produced by Geniotrigona thoracica (G. thoracica) from West Sumatera against extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli). In vitro antimicrobial assays were performed by well diffusion and serial dilution methods on six EEP concentrations: 100%, 50%, 25%, 12.5%, 6.25%, and 3.125%. No inhibitory zones were observed in the agar diffusion for any of the propolis concentrations tested. Similarly, neither the minimum inhibitory concentration (MIC) nor the bactericidal concentration (MBC) for each propolis concentration could be determined. These findings indicate that G. thoracica EEP has no antibacterial activity against ESBL-producing E. coli. On the other hand, phytochemical analyses revealed that the total phenolic, flavonoid, and alkaloid contents of G. thoracica EEP were 0.008 mg/mL, 0.015 g/g, and 0.084 g/g, respectively. These results suggest that G. thoracica propolis contains active compounds, which are thought to be components that contribute to its pharmacological activity.