Elicitor and precursor-induced approaches to enhance the in vitro production of L-DOPA from cell cultures of Mucuna pruriens

Abstract

Elicitation and precursor feeding are two important strategies in the in vitro techniques to enhance metabolite production to meet the demand of mankind. The secondary metabolites produced by the plants are extensively used in pharmaceutical, food and agro-chemical industries. One such metabolite is 3,4 dihydroxyphenylalanine ( L -DOPA) produced from Mucuna pruriens (L.) DC. is used since ancient times to treat Parkinson’s disease. Though all parts produce L -DOPA, the seed has the highest quantity. To overcome the extensive usage of the natural sources whose growth and metabolite production is highly dependent on edaphic and ecological factors, in vitro techniques like establishing cell culture for continuous production of metabolites, precursor feeding and elicitation of cell cultures to enhance the metabolite production has been reported in the present study. Callus was developed from the in vitro leaf explant and cell suspension culture was established in the liquid Murashige and Skoog’s medium fortified with 0.5 mg/L picloram . Amino acid precursors like tyrosine , phenylalanine and chemical elicitors like methyl jasmonate , salicylic acid, sodium nitroprusside and silver nitrate were exposed to cell cultures for different periods (3, 6 and 9 days respectively). The precursors showed a better response in enhancing both the biomass and L -DOPA when compared to the elicitors. 500 and 1000 mg/L tyrosine showed a 1.6- and an 8.1-fold increase in biomass and L -DOPA production respectively when supplemented with MS media . However, though all the elicitors enhanced the L -DOPA production by 1.1–3.3-folds they did not show much significant increase in biomass. Precursor feeding approaches enhanced the metabolite considerably more than the elicitor treatment. Based on the productivity (Biomass × L -DOPA conc.) precursors like Tyrosine>Phenylalanine and elicitors like Sodium nitroprusside>Silver nitrate>Methyl jasmonate>Salicylic acid showed better response. • First report to elicit L -DOPA from Mucuna pruriens cell cultures using Phenylalanine. • A highest of 8.1-fold increase of L -DOPA was achieved using Tyrosine as a precursor. • Among the elicitors, a 3.3-fold increase of L -DOPA was seen using sodium nitroprusside. • On the basis of productivity, Tyrosine and sodium nitroprusside showed best results.