Many antimicrobial drugs are going to become resistant to different pathogens, so the discovery of new antimicrobial products is an important public health concern. Nardostachys jatamansi (D. Don) DC (Valerianaceae) is an important traditional herbal medicine used as tranquillizer. CNS sedative. antiepilepsy, cardiotonic. diuretic, heptatonic, analgesic and in boils, itch and eye diseases. Carbonic anhydrase inhibitors are the potential source of treatment in case of glaucoma, hypertension and epilepsy and also used as diuretics. The aim of present study was to evaluate the antibacterial activity of methanolic extract of N. jatamansi against different bacterial strains and to estimate its carbonic anhydrase inhibitory potential. Antibacterial activity was evaluated by agar well diffusion assay and broth microdilution assay. The study revealed that N. jatamansi extract is sensitive to all tested bacterial strains. The zones of inhibition and MIC ranged from 8-22 at a concentration of 1-5 mg/mL and 0.3-0.6 mg/mL. respectively. Methanolic extract of N. jatamansi showed marked inhibition of carbonic anhydrase (IC 50 712.41 +/- 0.001 mu g/mL) when compared with standard acetazolamide. The results of the study suggest that N. jatamansi may be a valuable plant source of medicinally useful active compounds that can be helpful in bacterial infections and showed some relation with the traditional use of this plant in various diseases.