Cephalopods, especially Sepioteuthis lessoniana, are ecologically vital and economically important marine molluscs. This study investigates their hidden biomedical potential, focusing on chemical composition and antibacterial properties. Antibacterial assays against five pathogens showed inhibition zones for all except E. coli, indicating selective resistance. Bioactive compounds in squid extract were identified using UV-Vis and FTIR spectroscopy. Further analysis through FESEM, EDAX, and XRD revealed detailed microstructural and elemental profiles, emphasizing the chemical richness and antimicrobial potential of Sepioteuthis lessoniana extracts for biomedical applications. This multidisciplinary investigation unveils both the structural and functional attributes of bioactive compounds derived from S. lessoniana, establishing it as a promising candidate in the search for novel antimicrobial agents. In the face of rising global antibiotic resistance, marine bioprospecting emerges as a vital strategy for discovering new therapeutic options. S. lessoniana, with its rich chemical complexity and potent bioactivity, serves as a compelling example of the untapped potential within marine organisms. This study not only highlights its biomedical significance but also encourages further research into cephalopod chemistry as a foundation for innovative pharmaceutical developments and next-generation antimicrobial therapies.