Production of Taxol and its Analogues from Cell Cultures of Taxus wallichiana

Abstract

Callus cultures of Taxus wallichiana young leaves were established on Gamborg’s (B5) medium supplemented with dicamba (2 mg/l), benzyladenine (0.2 mg/l) and glutamine (292 mg/l). The cell cultures were initiated and maintained on B5 medium supplemented with NAA (1mg/l) and benzyladenine (0.02 mg/l). 0.25 B5 medium supplemented with 2,4-D (2 mg/l) and zeatin (0.5 mg/l) was found to be suitable as production medium. On HPLC analysis using photodiode array detector, the extracts from cell cultures showed the presence of taxanes, viz., taxol, deacetyl baccatin III (DAB) and baccatin III (BA). Precursors and growth retardants significantly improved the production of taxol, DAB and BA. Phenylalanine (1 mM), sodium benzoate (0.2mM), hippuric acid (1 mM) and leucine (1 mM) addition enhanced the accumulation of taxol (6, 4.5, 25 and 2 times, respectively), DAB (7, 9.5, 7.5 and 3 times, respectively) and BA (36, 16, 57 and 2 times, respectively) in cultures grown in production medium. Ancymidol (50µM) and 2-chloroethyl phosphonic acid (50µM) addition has shown significant increase in bioproduction of taxol (3 and 11 times, respectively), DAB (3 and 1.5 times, respectively) and BA (16 and 9 times, respectively). However, chlorocholine chloride (1mM) improved the production of DAB (2.5 times) and taxol (1.7 times) only.