Biotechnology Guided Production Of L-Dopa

Gajanand Engla², Deependra Singh¹, Prashant Jain¹ and V. K. Dixit¹*

¹Pharmaceutical Biotechnology Laboratory, Department of Pharmaceutical Sciences, Dr. H.S. Gour University, Sagar (India). ²Sagar Institute of Research and Technology, Pharmacy, Bhopal - 462 003 (India)

ABSTRACT: Transformed callus culture of Mucuna italies (Fabaceae) were obtained by infecting explants (epicotyls, hypocotyls and cotyledons) obtained from aseptically germinated seedlings and 4 weeks old callus of epicotyl on MS media supplemented with NAA (1.0 ppm) + BAP (1.0ppm) with Agrobacterium tumefaciens (MTCC-431, MTCC-2250 and MTCC-2251). The oncogenic strains had different level of virulence on different types of explants. The main difference was found in the nature of galls formed and in their subsequent morphological competence. The comparative callus induction efficiency of various strains was evaluated. Various transformed callus grew axenically in the absence of exogenous plant growth regulator in Murashige and Skoog's medium containing 3% w/v of sucrose. The best result was found with epicotyl explants with A. tumefaciens strain (MTCC-2250). The highest transfection efficiency was observed with MTCC 2250. MTCC 431 also showed satisfactory results, while MTCC 2251 failed to transfect any of explants. The suspension culture of transformed callus were initiated and maintained successfully. The gall and cell suspension obtained synthesizes various normal constituents of plant out of which, L-Dopa was isolated, identified and estimated. The transformed callus showed callus induction efficiency (85% epicotyl callus transfected with MTCC 2250), viability (84% callus and 76% cell suspension), growth index (4.92) and productivity of L-DOPA (4.37% by Cell DW in callus and 7.88% by Cell DW in cell suspension) and it was found higher than the intact Mucuna pruriens seeds (3.61% by Cell DW).

KEYWORDS: L-DOPA; callus culture; Agrobacterium tumefaciens; Mucuna sps

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