Research on medicinal plants is developing each day due to inborn phytochemicals, which can encourage the progress of novel drugs. Most plant-based phytochemicals have valuable effects on well-being. Among them, beetroot leaves (BL) are known for their therapeutic properties. Here, three solvents, namely, acetonitrile, ethanol, and water, and their combinations were developed for BL extraction and simultaneous assessment of phytochemical compounds and antioxidant and antifoodborne pathogen bacteria activities. By using the augmented simplex-centroid mixture design, 40.40% acetonitrile diluted in water at 38.74% and ethanol at 20.86% favored the recovery of 49.28 mg GAE/mL (total phenolic content (TPC)) and 0.314 mg QE/mL (total flavonoid content (TFC)), respectively. Acetonitrile diluted in water at 50% guarantees the best antioxidant activity, whereas the optimal predicted mixture for the highest antibacterial activity matches 24.58, 50.17, and 25.25% of acetonitrile, ethanol, and water, respectively. These extraction conditions ensured inhibition of
Staphylococcus aureus
,
Salmonella enterica
, and
Escherichia coli
, respectively, at 0.402, 0.497, and 0.207 mg/mL. Under optimized conditions, at three concentrations of BL, minimal inhibitory concentration (MIC), 2 × MIC, and 4 × MIC, a linear model was employed to investigate the inhibition behavior against the three tested bacteria. The early logarithmic growth phase of these bacteria illustrated the bactericidal effect of optimized extracted BL with a logarithmic growth phase inferior to 6 h. Therefore, BL extract at 4 × MIC, which corresponds to 1.608, 1.988, and 0.828 mg/mL, was more efficient against
S. aureus
,
S. enterica
, and
E. coli.