Impact of nutrient components on production of the phytoestrogens daidzein and genistein by hairy roots of Psoralea corylifolia.

Transformed hairy roots of Psoralea corylifolia were established by infection with Agrobacterium rhizogenes LBA 9402. The aim of this work was to elucidate the effects of media constituents on production of the phytoestrogenic isoflavones daidzein and genistein. A. rhizogenes strain LBA 9402 harboring Ri plasmid was used to transform stem segments of in vitro seedlings. The resultant hairy roots were confirmed by polymerase chain reaction (PCR) and exhibited Ri T-DNA. Transformed hairy root clones were cultured in Murashige and Skoog's (MS) medium altered with different concentrations of NH(4) (+) and NO(3) (-) and their growth and production of isoflavones were assessed. Biomass and productivity increased when MS medium was supplemented with NH(4) (+) and NO(3) (-) at a ratio of 20:10. Increased yield of daidzein was obtained when sucrose level in the culture medium increased, whereas decreased level of sucrose favored genistein production. The hairy roots produced the highest levels of daidzein (2.06% dry wt.) and genistein (0.37% dry wt.) in the presence of low concentrations of PO(4) (3-). Hairy roots secreted trace amounts of daidzein and genistein into the culture medium. The present results demonstrated that the productivity of daidzein was 2.2-fold more than that of untransformed roots.

Studied enhancement strategies for phytoestrogens production in shake flasks by suspension culture of Psoralea corylifolia.

This study proposed secondary metabolites incremental yield due to manipulation of nutrient components into the culture medium. To validate this, the effects of nutrients such as carbon, phosphate and nitrogen on growth and production of phytoestrogens daidzein and genistein by suspension cultures of Psoralea corylifolia was investigated for the first time. The maximum production of daidzein and genistein was achieved when sucrose and maltose used as a sole source of carbon. Suspension cell cultures enriched with sucrose (3%) stimulated accumulation of isoflavones daidzein (1.76% dry wt) and genistein (0.25% dry wt) compared to glucose, fructose and maltose. Sucrose feeding strategy significantly stimulated biomass growth and isoflavones (2.79% dry wt of daidzein and 0.32% dry wt of genistein) production rate. Reduced concentrations of phosphate (0.625 mM) promoted daidzein (1.89% dry wt) and genistein (0.26% dry wt) production by suspension cell cultures, whereas high amount (5mM) in medium was inhibited isoflavones production. It was observed that medium fortified with NH(4)(+) and NO(3)(-) alone inhibited production of isoflavones. The maximum production obtained of daidzein (2.20% dry wt) and genistein (0.29% dry wt) when medium comprised with NH(4)(+)/NO(3)(-) at ratio 20:40 mM as a nitrogen source. Similar nutrient components ratio when altered NH(4)(+)/NO(3)(-); 40:20mM) resulted in approximately 3-fold decrease in production. HPLC analysis revealed that suspension cells cultures leached out trace amount of daidzein and genistein into the culture medium.