Background: Corchorus olitorius L. (Family: Tiliaceae) or ‘Tossa Jute or Nalta Jute is an important fiber crop in India and Bangladesh. Its leaves are used as folk medicine, food source and have demulcent, diuretic, lactagogue, purgative, antidiabetic, anticancer, antioxidant, anti-inflammatory and tonic effects. The aim of the research is to evaluate the pharmacognostic characteristics and antimicrobial activity of Corchorus olitorius L. (C. olitorius). Materials and Methods: The pharmacognostic studies were carried out in terms of organoleptic, macroscopic, microscopic, physico-chemical evaluation, phytochemical screening, fluorescence analysis, TLC fingerprinting by standard procedures. Antibacterial activity against gram-negative bacteria E. coli was determined by agar well diffusion method. Results: The leaf of C. olitorius showed serrate margin, stipules at the base, and acute to acuminate apex. Leaves surface microscopy showed numerous prismatic calcium oxalate crystals and covering trichomes. The moisture content 6.6% w/w, ethanol and water-soluble extractive value 4 and 17.6% w/w, total ash 12% w/w, while acid insoluble and water-soluble ash value 2.2 and 2.0% w/w respectively were determined. Phytochemical screening showed the presence of flavonoids, polyphenols, phytosterols/triterpenoids, alkaloids, carbohydrates, tannins, and proteins. Thin layer chromatographic profiling of ethanolic extract was carried out which showed different Rf values. The antimicrobial test showed that aqueous extract has better antimicrobial activity around the tested bacteria (E. coli) than ethanolic extract. Conclusion: The results are helpful in the standardization of the leaf part of the plant C. olitorius. The aqueous leaf extract possessed antimicrobial properties. There is a need to isolate its constituents responsible for antimicrobial activity.