<p>A combined electrochemical oxidation and biological treatment process was studied for levofloxacin removal from water. An electrochemical oxidation process using graphite electrodes was employed to degrade levofloxacin (C₀ = 5&#xa0;mg/L), and biological treatment of residual levofloxacin and byproducts was conducted using an adapted <i>Enterobacter cloacae</i> strain, resulting in 90% total removal of levofloxacin after 5&#xa0;days. In contrast, biological treatment of 5&#xa0;mg/L levofloxacin with an adapted <i>Enterobacter cloacae</i> strain yielded merely 36% removal after 10&#xa0;days. Liquid chromatography–mass spectrometry analysis revealed that toxic intermediates generated during electrochemical oxidation were degraded by microbial processes. Enzymatic assays indicated the activity of manganese peroxidase and naphthalene dioxygenase, whereas the activities of laccase and horseradish peroxidase were not detected. A novel biodegradation pathway for levofloxacin was proposed. The two-stage approach is proposed as an energy-efficient strategy with reduced chemical demand and potential applicability to hospital or pharmaceutical effluents with high antibiotic loads.</p> Graphical abstract <p></p>