Datura metel: In vitro production of tropane alkaloids

Hairy root lines of Datura metel were established following infection of as eptic stem segments with Agrobocterium rhizogenes strain A4 and cultured in hormone-free B5 solid medium. The growth and production of hyoscyamine and scopolamine (mg/g dry wt.) of these root cultures was encouraged by using B5 liquid medium with half-strength salts. In these culture conditions, the capacity of the highest productive root line 25 to excrete scopolamine int o the culture medium rose from 8.7% to 70% when the permeabilizing agent Tw een 20 was added for 24 h to the medium, after 2 and 4 weeks of culture. Us ing an airlift bioreactor (41) with modifications in order to increase root anchorage, the Tween 20 treatment encouraged both growth and alkaloid prod uctivity of cultured Foot line 25. After 4 weeks their biomass yield was 2. 3 mg/l/day and 0.84 mg/l/day of scopolamine was produced (70% extracellular ). The scopolamine released into the culture medium was separated with an A mberlite XAD-2 column located in the media exit.