The present study deals with the chitin and chitosan obtained from tasar silkworm (Antheraea mylitta) spent pupae and characterized for physical characteristics by electron microscopy, infrared spectroscopy, thermo-gravimetric analysis, X-ray diffraction, elemental analysis, color and phenolic content. The molecular weight was determined by the viscometric method. In addition to these, the antioxidant, antimicrobial potential, cytotoxicity, and post-harvesting shelf-life of fruit was studied. The FESEM indicates that the surface of isolated chitosan was flat, smooth and nonporous. The FTIR spectra of tasar pupal chitosan exhibited similarity with other chitosan. TGA curve showed similar thermal stability, and elemental analysis showed the carbon: nitrogen ratio to be nearer to shrimp chitosan. The color analyzed by the colorimeter indicated less brightness and more color intensity. The water and fat binding capacity, antioxidant potential, and antimicrobial activity were also observed in prepared chitosan. Further, cells were viable in different tested concentrations of tasar pupal chitosan. Additionally, the chitosan coating application reduced the post-harvest damage and increased the shelf-life of the banana. The results suggest that the chitosan obtained from tasar spent pupae may open a new perspective in food and biomedical applications.