Mycobacterium tuberculosis (Mtb) is an opportunistic pathogen that has been responsible for the deadly Tuberculosis (TB). Mtb armors itself with a multitude of virulence factors that aid in establishing and progression of the infection. In recent years, Mtb has been shown to produce small molecules which collectively are termed as volatile organic compounds (VOCs) and have been detected in Mtb axenic culture and in the breath of TB patients. VOCs are produced by other pathogenic and non-pathogenic bacteria and have many functions including modulating growth and establishing relationship between host and environment. In this doctoral work, we studied the roles of three VOCs from Mtb. The role of VOCs as molecules influencing Mtb physiology especially of cell-to-cell signaling was assessed using M. smegmatis as a model. Also studied is the role of the VOCs as molecules mediating inter-species and inter-kingdom signaling using bacterial and fungal pathogens respectively. We found that VOCs have modulated biofilm formation and sliding motility in M. smegmatis. Resistance to rifampicin was enhanced upon VOC treatment. Sequence analysis of Mtb and M. smegmatis LuxR proteins (the classical quorum sensing machinery) was performed to assess if they are potential targets for VOCs. In the second part of our work, we showed that VOCs participate in cross-species cross-talk by modulating growth and virulence traits of P. aeruginosa, S. aureus, and C. albicans. Virulence traits like biofilm formation, secreted products like siderophores, pigment, hyphae formation, antibiotic resistance to gentamicin, penicillin, rifampicin, amphotericin B, caspofungin, and fluconazole, and motility were significantly altered with VOC exposure. Overall, Mtb VOCs have many physiological activities on M. smegmatis and possibly engage in bacterial signaling.