Triphala is a familiar ayurvedic formulation, commonly prescribed by most healthcare practitioners in India. It is an equiproportional mixture of fruits of three medicinal herbs, amalaki (Emblica officinalis), haritaki (Terminalia chebula) and bibhitaki (Terminalia bellerica). It exhibits anti-viral, anti-bacterial, anti-fungal and anti-allergic properties. Triphala and its constituents act as cardio-tonic, control blood pressure, improve blood circulation and reduce cholesterol levels. It also shows immune modulatory properties and helps in improving the body’s defense system. The aim of this study was to provide an overview of the antibacterial activities of the crude extracts of Triphala extracts (Aqueous, Methanol and Acetone) with special emphasis on its biochemical and molecular mechanisms of action against B.subtils. the antibacterial activities of triphala extracts against B.subtils were determined by agar well diffusion method, MTT Assay using Broth dilution method (With and without Triphala extract) used for biochemical analysis and RT-PCR for gene expression studies on selected metabolic genes of B.subtils which is important in stress condition (lex A, Lrg and Cid genes) were performed. The susceptibility test showed promising antibacterial activities of all the tested extracts against B.subtils. Among the tested extracts of triphala, Methanol exhibited the highest antibacterial activity and the highest levels of cytotoxic activity. In the gene expression studies of those selected candidates triphala have showed a capacity of up regulating all genes of interest, Cid is most expressed gene (5.6 fold) which is followed by, Lrg (4.8 fold) and Lex A (3.3 fold) respectively.When we compare each solvent, distinct extractants cause distinct genes moreexpressed than the other, lex A expressed more by methanol extract, whereas cid and Lrg expressed more in acetone and aqueous extracts respectively .Cid gene which is a critical effecter for bacterial lysis, may form membrane lesions through which endolysin occur ,muraytic enzyme escape the cytoplasm to attack the cell wall. When these genes expressed more they induce the release of muralytic enzymes from the cytoplasm and lead to membrane lesions and attack cell wall which results hole formation in the bacterial plasma membrane and finally cause to death of cell.