<p>The immense research on nanotechnology has opened up the way for a lot of new advancements which have paved the way for their applications in health biotechnology. Carbon nanotubes are rolled-up sheets of layers of carbon atoms. They mark tremendous applications in the medical as well as industrial fields. Carbon nanotubes are being synthesized by chemical and physical methods to date, which are not so eco-friendly. In the current research, carbon nanotubes were synthesized using <i>Bacillus subtilis</i> bacteria. In this study, <i>B. subtilis</i> is observed as a carbon nanotube synthesizer, a phenomenon previously unreported in scientific literature. Bacterial isolates were obtained using streak and spread plating, then characterized morphologically via Gram staining and genetically via 16S rRNA sequencing. Carbon nanotubes were characterized using ultraviolet-visible (UV–Vis) spectroscopy, scanning electron microscopy, zeta sizer analysis, and X-ray diffraction (XRD). The biosynthesized carbon nanotubes had a fusiform and long tube-like shape with a hollowed core encircled by many graphene layers, having an average size of 78.82&#xa0;nm. Bacterial isolate identity was confirmed by 16S rRNA gene sequence analysis. The homogeneity with other strains (<i>Bacillus velezensis</i> or <i>Bacillus halotolerans</i>) was 99%.The antimicrobial assay indicated that the compound exhibited no efficacy against the tested pathogens, <i>Staphylococcus aureus</i>, <i>Salmonella typhi</i>, and <i>Escherichia coli</i>, as evidenced by a complete absence of zones of inhibition. Consequently, these bacterially synthesized carbon nanotubes (CNTs) present a promising avenue for future research, particularly in exploring their practical applications in biomedical fields, such as targeted drug delivery and biosensing, as well as in environmental remediation.</p>