Abstract
The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period.
Highlights
Plants produce a wide range of secondary metabolites, which are used for a number of applications, such as pharmaceuticals, bio-pesticides, flavors, fragrances, colors and food additives
The biomass accumulation (FW and dry weight (DW)) and withanolides production in MS medium containing picloram, KN, L-glutamine and sucrose are shown in figure 1a and 1b
The biomass reached a maximum with a fresh weight of 14.72 g and a dry weight of 3.68 g on 28th day (Figure 1a)
Summary
Plants produce a wide range of secondary metabolites, which are used for a number of applications, such as pharmaceuticals, bio-pesticides, flavors, fragrances, colors and food additives. At present, these products have been obtained from plants growing in the wild or cultivated sources. Cell suspension culture offers a condensed biosynthetic cycle to study the growth and production kinetics within a short cultivation time (about 2–4 weeks) with an added advantage of tunability that can help to implement optimal conditions for the production of a number of high value medicinal compounds in good quantities [2,3]
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