Production of puerarin and isoflavones in cell suspension cultures of Pueraria lobata (Willd.): effects of medium supplementation with casein hydrolysate and coconut milk.

ABSTRACT

Callus induced from leaf explants of Pueraria lobata seedlings were suspended in Gamborg B5 medium supplemented with 1 mg l(-1) 2,4-dichlorophenoxyacetic acid, 1 mg l(-1) naphthalene acetic acid, 0.5 mg l(-1) kinetin and 30 g l(-1) sucrose. The effects of coconut milk and casein hydrolysate (CH) on cell growth and yields of puerarin and isoflavones in cells suspension were studied. The contents of total isoflavones and puerarin in suspension cultures were determined by spectrophotometry and HPLC. Coconut milk (10%, filter sterilized) decreased the growth of cell cultures and the accumulation of total isoflavones, while 0.2% CH promoted the growth of cell cultures and the accumulation and release of puerarin and total isoflavones. The total yield of puerarin and isoflavones were 34% and 40.8% higher than in the control, respectively. The optimum medium for cell cultures of leaves of P. lobata seedlings was B5 liquid medium supplemented with 2% sucrose, 1.0 mg l(-1) 2,4-D, 1.0 mg l(-1) NAA, 0.5 mg l(-1) kinetin and 20 mg l(-1) CH. The procedure use is a potentially useful for the production of isoflavones.