The purpose of the present study is to determine the antibacterial activity of Piperine extracted from Piper nigrum and its synergistic effect with antibiotic drug (Ciprofloxacin) against Escherichia coli and Bacillus subtilis. The research targets expensive, toxic drug inputs that exhibit poor bioavailability with easily available, non-toxic, natural plant extracts for similar/improved outcomes. Piperine was extracted from Piper nigrum. The antibacterial activities of this extract were evaluated using the drug inhibition zone formation assay. The inhibitory zones were recorded in millimeters. Combinations of Ciprofloxacin and Piperine have shown synergistic activity against both the organisms. Extracted Piperine from Piper nigrum was active against Bacillus subtilis and E. coli even at a very low concentration (20 μg/mL) in combination with Ciprofloxacin.