Indian Journal of Science and Technology

Abstract

 In an attempt to increase the productivity of the pharmaceutical compound canthin-6-one from cell suspension culture of Brucea javanica, a high cell density culture and improved culture conditions have been investigated with focus on culture conditions and stages of growth. The highest yield of canthin-6-one, 26.72 mg g-l dry cell, was produced intracellular in 50 g L-1 cell mass. The established cell suspensions were harvested at the age of 40 days and placed into MS medium containing 1.0 mg L-1 2,4-D, 1.0 mg L-1 NAA, 0.1 mg L-1 Kinetin and 10 mg L-1 tryptophan for 33-40 days, approximately when the cultures were at their late stationary phase of growth. Addition of 20 mg L-1 tryptophan did not show a significant difference in canthin-6-one production (p>0.05). Higher than 10 mg L-1 of tryptophan showed adverse effect on the concentration of canthin-6- one alkaloid (p=0.01). Qualitative analysis of chloroform-extracts of dried cells from suspension cultures revealed spots on TLC; and the identity of canthin-6-one was established by comparison of its Rf value (Rf=0.37), its colour reaction with Dragendorff reagent and ammonium sulphate and its behaviour in UV with an authentic sample. Quantification of canthin-6-one alkaloid using TLCdensitometry scanner in response to different concentration of tryptophan as a precursor showed a concentration-dependent response. The production of canthin-6-one at 26.72 mg g-1 dried cell weight in this study, establishes a methodology of an improved procedure compared to those previously reported and offers an opportunity for production of canthin-6-one alkaloid at an industrial level.

Keywords: Canthin-6-One, cell suspension, Brucea javanica, secondary metabolites