title: Cell line selection combined with jasmonic acid elicitation enhance camptothecin production in cell suspension cultures of Ophiorrhiza mungos L.
abstract: Ophiorrhiza mungos is a herbaceous medicinal plant which contains a quinoline alkaloid, camptothecin (CPT), an anticancer compound. A high-yielding cell line, O. mungos cell line-3 (OMC3) was selected from cell suspension cultures of O. mungos using cell aggregate cloning method and established cell suspension culture. OMC3 cell suspension produced significantly high biomass (9.25 ± 1.3 g/flask fresh weight (FW)) and CPT yield (0.095 ± 0.002 mg g-1dry weight (DW)) compared with the original cell suspension. Inoculum size of OMC3 cell suspension culture was optimised as 14 g L-1. Media optimisation has shown that 5 % (w/v) sucrose and an increased ammonium/nitrate concentration of 40/20 mM favoured CPT production, whereas 3 % (w/v) sucrose, an ammonium/nitrate concentration of 20/40 mM and 1.25 mM of phosphate favoured biomass accumulation. Jasmonic acid, chitin and salicylic acid was used to elicit CPT production in the original cell suspension culture and achieved significantly high CPT production with jasmonic acid (JA) elicitation. Further, OMC3 cell suspension culture was elicited with JA (50 μM) and obtained 1.12 ± 0.08 mg g-1DW CPT and 9.52 ± 1.4 g/flask FW (190.4 g L-1FW). The combination of cell line selection and elicitation has produced 18.66-fold increases in CPT production together with significantly high biomass yield. The study is helpful in the scale-up studies of O. mungos cell suspension culture in suitable bioreactor systems for the production of CPT.
authors: Division of Biotechnology and Bioinformatics, Jawaharlal Nehru Tropical Botanic Garden & Research Institute, Palode, Thiruvananthapuram, Kerala, 695 562, India.
authors: S
authors: S
authors: Deepthi
authors: Division of Biotechnology and Bioinformatics, Jawaharlal Nehru Tropical Botanic Garden & Research Institute, Palode, Thiruvananthapuram, Kerala, 695 562, India. bioproduction09@gmail.com.
authors: K
authors: K
authors: Satheeshkumar
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism*; D018533:Biomass; D002166:Camptothecin / Q000378:metabolism*; D018929:Cell Culture Techniques; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D003517:Cyclopentanes / Q000378:metabolism*; D054883:Oxylipins / Q000378:metabolism*; D010937:Plant Growth Regulators / Q000378:metabolism*; D019666:Rubiaceae / Q000378:metabolism*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D003470:Culture Media; D003517:Cyclopentanes; D054883:Oxylipins; D010937:Plant Growth Regulators; C011006:jasmonic acid; D002166:Camptothecin
keywords: Camptothecin; Cell aggregate cloning; Cell line; Elicitation; Inoculum size; Jasmonic acid