Enhanced taxol production and release in Taxus chinensis cell suspension cultures with selected organic solvents and sucrose feeding

Suspension culture of Taxus chinensis cells was carried out in aqueous-orga nic two-phase systems for the production and in situ solvent extraction of taxol (paclitaxel). Three organic solvents, hexadecane, decanol, and dibuty lphthalate, were tested at 5-20% (v/v) in the culture liquid. All of these solvents stimulated taxol release and the yield per cell, though decanol an d higher concentrations of the other two solvents depressed biomass growth significantly. Ten percent dibutylphthalate was the optimal solvent for imp roving taxol production and release with minimal cell growth inhibition. Th e time of solvent addition to the culture also affected taxol production, w ith the addition during the late-log growth phase being most favorable. By feeding sucrose to the culture near the stationary growth phase, the cell g rowth and taxol production period was extended from 27 to 42 days. The comb ining of the two-phase culture and sucrose feeding increased the taxol yiel d by about 6-fold compared with the single-phase batch culture, to 36.0 +/- 3.5 mg/L, with up to 63% taxol released. This study shows that in situ sol vent extraction combined with nutrient feeding is an effective process stra tegy for production and recovery of secondary metabolites in plant cell sus pension culture.