The bioprocess methodology for the enhanced production of phytochemicals using plant cultures predominantly depends on the elicitors and precursors. The drug L-DOPA (anti-Parkinson's drug) (L-3, 4-dihydroxyphenylalanine) has been chiefly isolated from the popular medicinal plant species Mucuna. The present study has been carried out to enhance the production of L-DOPA and other secondary metabolites in the presence of the optimized concentration of growth promoters, elicitors and precursor's in vitro callus culture of Mucuna imbricata. Results revealed that there is a positive influence of Methyl Jasmonate (MJC) and Salicylic Acid (SAC) on the accumulation of L-DOPA and other secondary metabolites (Nutritional and anti-nutritional factors), which enhances their production by more than two folds. The highest accumulation of L-DOPA was noticed in the elicitation of 1 mM MJC (4.3 ± 0.4 %) followed by 1 mM SAC (3.4 ± 0.08 %) on 45th and 30th days respectively. Results also illustrated a significant increase of callus biomass (dry and moist weight) as well as total phenolics, flavonoids and antioxidants values in the elicited callus from day 30th to day 45th of explant inoculation which indicates a synergistic interaction of auxin, cytokinins with elicitors and precursors respectively. Additionally, results revealed that improved accumulation of carbohydrates was positively correlated with an increase in L-DOPA content in the elicited callus. Hence the protocol formulated delivers an effective alternative for mass production of L-DOPA from callus culture with improved metabolite content.