Introduction and Aim: DNA gyrase is a class of Type II Topoisomerases that plays an important role in bacterial viability. It is found in all bacteria and is involved in replication, repair, recombination, and DNA transcription. Negative supercoiling of bacterial DNA by DNA gyr B is essential in replication which further influences all the metabolic activities. Staphylococcus aureus (ATCC 25923) is one of the pathogens that can modify its genome easily under multidrug resistance. In this study, the activity of medicinal compounds to inhibit DNA gyrB is explored. Plant species Solanum nigrum, Vitex negundo, and Euphorbia hirta were studied for the potential plant-based molecules. The compounds alkaloids, glycosides, flavonoids, and terpenoids were considered to have high-potential targets. The study focuses on DNA gyrase as a target and shows insights into future drug development. The research focuses on the discovery of novel plant-based therapeutic compounds to target DNA gyrase B activity. Methods and Materials: Phytochemical screening was performed to study the medication options that could inhibit DNA gyrB. Phytochemicals were determined using GC-MS. Results: Utilizing GC MS and FT-IR analysis, the phytochemical constituents of Solanum nigrum, Vitex negundo, and Euphorbia hirta were discovered. It will be simpler to do a follow-up study on discovering bioactive compounds and evaluating their effectiveness in inhibiting DNA gyrB with the help of this preliminary data from the analytical procedures. Conclusion: There are countless applications for the phytochemicals that medicinal plants produce. Staphylococcus aureus will be stopped by DNA gyrB inhibition. The study employs DNA gyrase as its target and provides information on potential therapeutic targets. The goal of the study is to identify innovative plant-based medicinal molecules that specifically target DNA gyrase B activity.