ABSTRACT
The aryl tetralin lignans are synthesized by Podophyllum sps. and are in great demand worldwide due to their use in synthesis of topoisomerase inhibitors. However, the sustained production of these aryl tetralin lignans requires large-scale harvesting from the natural environments, which has resulted in the plant-endangered status. In view of the difficulties in their total chemical synthesis, cultivation and failure of metabolic engineering approaches, there is a need to search for alternative sources of production of aryl tetralin lignans. We unequivocally established the methodology for isolation, identification, and characterization of a novel fungal endophyte (Trametes hirsuta) that produces aryl tetralin lignans consistently as shown by HPLC, LC-MS, LC/MS-MS and (1)H NMR. The lignans produced by the microorganism are biologically active, and exhibit potent antioxidant, anticancer and radioprotective properties. This strategy promises to improve the production of these therapeutically important compounds at lower costs.
Subject(s)
Fungi/isolation & purification , Lignans/chemistry , Podophyllotoxin/chemistry , Tetrahydronaphthalenes/chemistry , Cell Fractionation , Chromatography, High Pressure Liquid , Fungi/cytology , Kinetics , Lipid Peroxidation/radiation effects , Models, Biological , Plant Cells , Plant Structures/cytology , Plant Structures/metabolism , Plants/metabolism , Podophyllotoxin/metabolism , Podophyllum/cytology , Podophyllum/metabolism , Spectrometry, Mass, Electrospray IonizationABSTRACT
The effect of major medium ingredients (sugar, nitrogen source and phosphate) in Podophyllum hexandrum suspension cultures was investigated in order to increase the production of podophyllotoxin, the raw material in the synthesis of anticancer drugs. Amongst B5, Eriksson, MS, Nitsch, Street and White's medium, MS medium resulted in high growth and podophyllotoxin accumulation. The optimum level of nitrogen was found to be 60 mM, with a combination of ammonium salts and nitrate in the ratio of 1:2. The highest level of podophyllotoxin was obtained at 60 g glucose/l and at 1.25 mM phosphate after 30 days. Statistical design was adopted to determine the optimum levels of the parameters for cell growth and podophyllotoxin production.
Subject(s)
Antineoplastic Agents, Phytogenic/biosynthesis , Podophyllotoxin/biosynthesis , Podophyllum/metabolism , Biotechnology , Cells, Cultured , Culture Media , Glucose/metabolism , Nitrogen/metabolism , Phosphates/metabolism , Plant Roots/cytology , Plants, Medicinal , Podophyllotoxin/analysis , Podophyllum/cytology , Podophyllum/growth & developmentABSTRACT
Podophyllotoxin, a potent chemotherapeutic agent is obtained from Podophyllum hexandrum Royle. Embryos of P. hexandrum were transformed using different strains of Agrobacterium rhizogenes viz. A4, 15834, K599. Transformed nature of the calli was ascertained and the cultures were further maintained as individual clones. HPLG analysis of transformed cultures depicted a three-fold increase in podophyllotoxin content in comparison to controls.
