Production of Podophyllotoxin Using Cross‐Species Coculture of Linum flavum Hairy Roots and Podophyllum hexandrum Cell Suspensions

Abstract

Novel cross-species coculture systems using Linum flavum hairy roots and Podophyllum hexandrum cell suspensions were applied for in vitro production of podophyllotoxin. The hairy roots and suspensions were cocultured in Linsmaier and Skoog medium in dual shake flasks and dual bioreactors. In separate experiments, coniferin feeding was shown to be an effective strategy for increasing the accumulation of podophyllotoxin in P. hexandrum suspensions. Because roots of L. flavum are a natural source of coniferin, hairy roots of this species were used in coculture with P. hexandrum to provide an in situ supply of coniferin. Compared with P. hexandrum suspensions cultured alone in shake flasks or bioreactors, podophyllotoxin concentrations in cocultured P. hexandrum cells were increased by 240% and 72% in dual shake flask and dual bioreactor systems, respectively. The availability and stability of coniferin in the medium are the most likely factors limiting podophyllotoxin synthesis in coculture. Intensification of the coculture process is required to further improve total podophyllotoxin accumulation on a volumetric basis.

A Review on Hemisynthesis, Biosynthesis, Biological Activities, Mode of Action, and Structure-Activity Relationship of Podophyllotoxins: 2003- 2007

“…Compared with P. hexandrum suspensions cultured alone, significant increases in PPT production were obtained using a range of coculture systems and operating conditions in shake flasks and dual bioreactors. These results demonstrated the feasibility of crossspecies coculture of hairy roots and cell suspensions for improved production of plant secondary metabolites [87].…”

“…Podophyllotoxin (PPT, 1), one of the well-known naturally occurring aryltetralin lignans, is extracted as the main component from the roots and rhizomes of Podophyllum species such as P. hexandrum and P. peltatum [1], whose medicinal properties, such as cathartic, antirheumatic, antiviral and antitumor activity, have been well-recognized for centuries [2]. But attempts to use PPT in the treatment of human neoplasia were mostly unsuccessful due to the complicated side effects such as nausea, vomiting, and damage of normal tissues, etc.…”

A comprehensive review on the phytochemical and pharmacological aspects of Podophyllum hexandrum: a high value medicinal plant

“…In this case, the coculture system can prove to be a judicious alternative as has been reported for the production of podophyllotoxin by Podophyllum hexandrum cell suspensions, synthesis of which is limited by the availability of coniferin. When Podophyllum hexandrum cells are cultured together with Linum flavum hairy roots, which produce and secrete coniferin, the production of podophyllotoxin was increased by 240% in the dual shaker flask system [57]. Hairy root cultures are genetically more stable and superior in their metabolic performance over suspension cultures of the cell line but scaling up hairy roots to industrial levels poses a great challenge at the moment and efficiency of the scaling up systems still needs optimization before industrial exploitation becomes valuable.…”

Hairy root biotechnology—indicative timeline to understand missing links and future outlook

“…Another judicious alternative and relatively simple method is the co-cultured hairy root systems. In such systems, hairy roots, together with a different in vitro system that produce and secret the compound which can act directly act as a precursor to metabolite synthesis in hairy roots or elicitate the root tissues to produce the metabolite, can be cultured (Lin et al 2003;Wu et al 2007). In the same context, precursor feeding in association with elicitation provided significant results in HRCs (Shinde et al 2009;Verma et al 2014).…”