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Evaluation of nutrient uptake and physical parameters on cell biomass growth and production of spilanthol in suspension cultures of Spilanthes acmella Murr.

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Abstract

Spilanthes acmella Murr. has a plethora of highly valuable biologically active compounds and has been listed as one of the important medicinal plants of the world. However, no perceptible biotechnological advances have been made for this genus to exploit or enhance its utility. To nullify the effect of seasonal variations, the present report is the first attempt to establish in vitro cell suspension cultures and to evaluate the production of spilanthol from them, a key component of the plant responsible for most of its pharmaceutical activities. The study examined the biomass growth in relation to the consumption of major nutrients and sucrose, agitation speed and dynamic change in pH. Results indicated that the consumption of phosphate resulted in the onset of decline phase in cultures. Spilanthol production was observed to be growth associated and maximum production occurred on the 15th day. Among the carbon sources, the highest production of spilanthol as 91.4 µg g−1 DW was recorded in the medium supplemented with sucrose, followed by glucose which produced 56.8 µg g−1 DW of spilanthol. Spilanthol could not be detected in fructose containing medium. Maximum viable cultures were obtained at a rotation speed of cells at 120 rpm. This study signifies the utility of Spilanthes suspension cultures for biosynthesis and constant production of spilanthol, throughout the year. The results of present study are useful for further scale-up process.

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Abbreviations

BAP:

N 6-Benzylaminopurine

2,4-D:

2,4-Dichlorophenoxyacetic acid

IAA:

Indole-3-acetic acid

Kinetin:

N 6-furfuryladenine

NAA:

α-Naphthaleneacetic acid

pCPA:

para-Chlorophenoxyacetic acid

TDZ:

Thidiazuron

References

  1. 1.

    Dörnenburg H, Knorr D (1995) Strategies for the improvement of secondary metabolite production in plant cell cultures. Enzym Microb Technol 17:674–684

  2. 2.

    Anonymous (1989) The wealth of India: a dictionary of Indian raw materials and industrial products. Council of Scientific and Industrial Research, New Delhi, vol 10, pp 11–12

  3. 3.

    Jondiko IJO (1986) A mosquito larvicide in Spilanthes mauritiana. Phytochemistry 25:2289–2290

  4. 4.

    Ramsewak RS, Erickson AJ, Nair MG (1999) Bioactive N-isobutylamides from the flower buds of Spilanthes acmella. Phytochemistry 51:729–732

  5. 5.

    Pandey V, Agrawal V (2009) Efficient micropropagation protocol of Spilanthes acmella L. possessing strong antimalarial activity. In Vitro Cell Dev Biol Plant 45:491–499

  6. 6.

    Guiotto P, Woelkart K, Grabnar I, Voinovich D, Perissutti B, Invernizzi S, Granzotto M, Bauer R (2008) Pharmacokinetics and immunomodulatory effects of phytotherapeutic lozenges (bonbons) with Echinacea purpurea extract. Phytomedicine 15:547–554

  7. 7.

    Matthias A, Connellan P, Thompson D, Bone KM, Lehmann RP (2008) Acute immune-modulatory effects of bioavailable Echinacea alkamides. Planta Med 74:1008–1009

  8. 8.

    Prachayasittikul S, Suphapong S, Worachartcheewan A, Lawung R, Ruchirawat S, Prachayasittikul V (2009) Bioactive metabolites from Spilanthes acmella Murr. Molecules 14:850–867

  9. 9.

    Adler RJ (2006) Compositions for the acute and/or long term treatment of periodontal diseases using herb extracts. Chem Abstr 145:14791. Eur Patent WO 2006059196

  10. 10.

    Khadir HA, Zakaria MB, Ketchil AA, Azirum MS (1989) Toxicity and electrophysiological effects of Spilanthes acmella Murr. extracts on Periplaneta americana L. Pesticide Sci 25:329–335

  11. 11.

    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497

  12. 12.

    Trevelyan WE, Harrison JS (1952) Studies on yeast metabolism 1. Fractionation and microdetermination of cell carbohydrates. Biochem J 50:298–303

  13. 13.

    Nigra HM, Alvarez MA, Giulietti AM (1990) Effect of carbon and nitrogen sources on growth and solasodine production in batch suspension cultures of Solanum eleagnifolium Cav. Plant Cell Tiss Org 21:55–60

  14. 14.

    Boonen J, Baert B, Roche N, Burvenich C, De Spiegeleer B (2010) Transdermal behaviour of the N-alkylamide spilanthol (affinin) from Spilanthes acmella (Compositae) extracts. J Ethnopharmacol 127:77–84

  15. 15.

    Cech NB, Tutor K, Doty BA, Spelman K, Sasagawa M, Raner GM, Wenner CA (2006) Liver enzyme-mediated oxidation of Echinacea purpurea alkylamides: production of novel metabolites and changes in immunomodulatory activity. Planta Med 72:1372–1377

  16. 16.

    Srivastava P, Sisodia V, Chaturvedi R (2011) Effect of culture conditions on synthesis of triterpenoids in suspension cultures of Lantana camara L. Bioproc Biosyst Eng 34:75–80

  17. 17.

    Kochan E, Chmiel A (2011) Dynamics of ginsenoside biosynthesis in suspension culture of Panax quinquefolium. Acta Physiol Plant 33:911–915

  18. 18.

    Luedeking R, Piret EL (1959) A kinetic study of the lactic acid fermentation: batch process at controlled pH. J Biochem Microbiol Technol Eng 1:393–431

  19. 19.

    Zhang Y-H, Zhong J-J, Yu J-T (1996) Enhancement of ginseng saponin production in suspension cultures of Panax notoginseng: manipulation of medium sucrose. J Biotechnol 51:49–56

  20. 20.

    Fett-Neto AG, Zhang WY, Dicosmo F (2004) Kinetics of taxol production, growth and nutrient uptake in cell suspensions of Taxus cuspidate. Biotechnol Bioeng 44:205–210

  21. 21.

    Prakash G, Srivastava AK (2007) Azadirachtin production in stirred tank reactors by Azadirachta indica suspension culture. Process Biochem 42:93–97

  22. 22.

    Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signalling in plants conserved and novel mechanisms. Ann Rev Plant Biol 57:675–709

  23. 23.

    Wang Y, Weathers PJ (2007) Sugars proportionately affect artemisinin production. Plant Cell Rep 26:1073–1081

  24. 24.

    Masuda H, Takahashi T, Sugawara S (1988) Acid and alkaline invertases in suspension cultures of sugar beet cells. Plant Physiol 86:312–317

  25. 25.

    Bondarev N, Reshetnyak O, Nosov A (2003) Effects of nutrient medium composition on development of Stevia rebaudiana shoots cultivated in the roller bioreactor and their production of steviol glycosides. Plant Sci 165:845–850

  26. 26.

    Nagella P, Murthy HN (2010) Establishment of cell suspension cultures of Withania somnifera for the production of with anolide A. Bioresour Technol 101:6735–6739

  27. 27.

    Su WW (1995) Bioprocessing technology for plant cell suspension cultures. Appl Biochem Biotech 50:189–230

  28. 28.

    Bonga JM, Von Aderkas P (1992) In vitro culture of trees. Kluwer Academic Publishers, Boston

  29. 29.

    Kretzschmar FS, JrCJF Oliveira, Braga MR (2007) Differential sugar uptake by cell suspension cultures of Rudgea jasminoides, a tropical woody Rubiaceae. In Vitro Cell Dev Biol Plant 43:71–78

  30. 30.

    Chattopadhyay S, Farkya S, Srivastava AK, Bisaria VS (2002) Bioprocess considerations for production of secondary metabolites by plant cell suspension cultures. Biotechnol Bioproc E 7:138–149

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Correspondence to Rakhi Chaturvedi.

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Singh, M., Chaturvedi, R. Evaluation of nutrient uptake and physical parameters on cell biomass growth and production of spilanthol in suspension cultures of Spilanthes acmella Murr.. Bioprocess Biosyst Eng 35, 943–951 (2012). https://doi.org/10.1007/s00449-012-0679-3

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Keywords

  • Agitation speed
  • Batch kinetics
  • Cell viability
  • Spilanthol
  • Spilanthes
  • Suspension cultures