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82 ± 1.46 for enzyme-substrate l-Asn, as obtained through MD simulations. Subsequently, fluorescence quenching and differential scanning calorimetry experiments validated the in silico data. The observance of inhibition of NgA activity at micromolar drug concentrations further strengthened our findings. Conclusive evidence came from the cell viability assays where these drugs were found to impede the growth of N. gonorrhoeae culture effectively. Thus, our study establishes l-asparaginase as a new molecular target against gonococcal in