Indian Journal of Science and Technology
Year: 2008, Volume: 1, Issue: 7, Pages: 1-6
ME.Wagih1 , G.Alam2 , S.Wiryowidagdo2 and K.Attia3
1 Department of Biology, Faculty of Science, University of New Brunswick, Saint John, NB E2L 4L5, Canada.
2Department of Pharmacy, Faculty of Natural Sciences, Hasanuddin University, Indonesia.
3Institute of Genetic Engineering, Fudan University, Shanghai, 200433, China.
*Author for the correspondence:
Department of Biology, Faculty of Science
University of New Brunswick, Saint John, NB E2L 4L5, Canada.
E-mail: [email protected]
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
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