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In Vitro Azadirachtin Production by Hairy Root Cultivation of Azadirachta indica in Nutrient Mist Bioreactor

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Abstract

Azadirachtin, a well-known biopesticide is a secondary metabolite conventionally extracted from the seeds of Azadirachta indica. The present study involved in vitro azadirachtin production by developing hairy roots of A. indica via Agrobacterium rhizogenes-mediated transformation of A. indica explants. Liquid culture of hairy roots was established in shake flask to study the kinetics of growth and azadirachtin production. A biomass production of 13.3 g/L dry weight (specific growth rate of 0.7 day−1) was obtained after 25 days of cultivation period with an azadirachtin yield of 3.3 mg/g root biomass. To overcome the mass transfer limitation in conventionally used liquid-phase reactors, batch cultivation of hairy roots was carried out in gas-phase reactors (nutrient spray and nutrient mist bioreactor) to investigate the possible scale-up of A. indica hairy root culture. The nano-size nutrient mist particles generated from the nozzle of the nutrient mist bioreactor could penetrate till the inner core of the inoculated root matrix, facilitating uniform growth during high-density cultivation of hairy roots. A biomass production of 9.8 g/L dry weight with azadirachtin accumulation of 2.8 mg/g biomass (27.4 mg/L) could be achieved in 25 days of batch cultivation period, which was equivalent to a volumetric productivity of 1.09 mg/L per day of azadirachtin.

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References

  1. Prakash, G., Bhojwani, S. S., & Srivastava, A. K. (2002). Biotechnology and Bioprocess Engineering, 7, 185–193.

    Article  CAS  Google Scholar 

  2. Srivastava, S., & Srivastava, A. K. (2008). In K. G. Ramawat & J. M. Merillon (Eds.), Biotechnology: bioactive molecules and medicinal plants. Germany: Springer.

    Google Scholar 

  3. Prakash, G., Emmannuel, C. J. S. K., & Srivastava, A. K. (2005). Biotechnology and Bioprocess Engineering, 10, 198–204.

    Article  CAS  Google Scholar 

  4. Prakash, G., & Srivastava, A. K. (2005). Process Biochemistry, 40, 3795–3800.

    Article  CAS  Google Scholar 

  5. Prakash, G., & Srivastava, A. K. (2006). Biochemical Engineering Journal, 29, 62–68.

    Article  CAS  Google Scholar 

  6. Prakash, G., & Srivastava, A. K. (2007). Process Biochemistry, 42, 93–97.

    Article  CAS  Google Scholar 

  7. Prakash, G., & Srivastava, A. K. (2008). Biochemical Engineering Journal, 40, 218–226.

    Article  CAS  Google Scholar 

  8. Giri, A., & Narasu, M. L. (2000). Biotechnology Advances, 18, 1–22.

    Article  CAS  Google Scholar 

  9. Srivastava, S., & Srivastava, A. K. (2007). Critical Reviews in Biotechnology, 27, 29–43.

    Article  CAS  Google Scholar 

  10. Allan, E., Eeswara, J., Jarvis, A., Mordue (Luntz), A., Morgan, E., & Stuchbury, T. (2002). Plant Cell Report, 21, 374–379.

    Article  CAS  Google Scholar 

  11. Satdive, R. K., Fulzele, D. P., & Eapen, S. (2007). Journal of Biotechnology, 128, 281–289.

    Article  CAS  Google Scholar 

  12. Carvalho, E. B., Holihan, S., Pearsall, B., & Curtis, W. R. (1997). In P. Doran (Ed.), Hairy roots: effect of root morphology on reactor design and operation for production of chemicals (pp. 151–167). Harwood Academic UK: Gordon and Breach.

    Google Scholar 

  13. Wyslouzil, B. E., Waterbury, R. G., & Weathers, P. J. (2000). Biotechnology and Bioengineering, 72, 143–150.

    Article  Google Scholar 

  14. Asplund, P. T., & Curtis, W. R. (2001). Biotechnology Progress, 17, 481–489.

    Article  CAS  Google Scholar 

  15. Kanokwaree, K., & Doran, P. M. (1997). Biotechnology and Bioengineering, 55, 520–526.

    Article  CAS  Google Scholar 

  16. Murashige, T., & Skoog, F. (1962). Physiology Plant, 15, 473–497.

    Article  CAS  Google Scholar 

  17. Gamborg, O. L., Miller, R. A., & Ojima, K. (1968). Experimental Cell Research, 50, 151–158.

    Article  CAS  Google Scholar 

  18. Srivastava, S. & Srivastava, A. K. (2006). Proceedings of national conference CHEMCON-06, Ankaleshwar, India.

  19. Srivastava, S., Harsh, S., & Srivastava, A. K. (2008). Process Biochemistry, 43, 1121–1123.

    Article  CAS  Google Scholar 

  20. Dubois, M., Gilf, K. A., Hamilton, J. K., Roberts, P. A., & Smith, F. (1956). Analytical Chemistry, 28, 350–356.

    Article  CAS  Google Scholar 

  21. Yuan, Y. J., Li, C., Hu, Z. D., & Wu, J. C. (2002). Enzyme and Microbial Technology, 30, 774–778.

    Article  CAS  Google Scholar 

  22. Eibl, R., & Eibl, D. (2002). In K. M. Oksman-Caldentey & W. H. Barz (Eds.), Plant biotechnology and transgenic plants (pp. 163–199). New York: Marcel Dekker.

    Google Scholar 

  23. Kim, Y. H., & Yoo, Y. J. (1993). Biotechnology Techniques, 7, 859–862.

    Article  CAS  Google Scholar 

  24. Buitelaar, R. N., Langenhoff, A. A. N., Heidstra, R., & Tramper, J. (1991). Enzyme and Microbial Technology, 13, 487–494.

    Article  CAS  Google Scholar 

  25. Pavlov, A., & Bley, T. (2006). Process Biochemistry, 41, 848–852.

    Article  CAS  Google Scholar 

  26. McKelvey, S. A., Gehrig, J. A., Hollar, K. A., & Curtis, W. R. (1993). Biotechnology Progress, 9, 317–322.

    Article  CAS  Google Scholar 

  27. Kim, Y. J., Weathers, P. J., & Wyslouzil, B. E. (2002). Biotechnology and Bioengineering, 80, 454–464.

    Article  CAS  Google Scholar 

  28. Ramakrishnan, D., & Curtis, W. R. (2004). Biotechnology and Bioengineering, 88, 248–260.

    Article  CAS  Google Scholar 

  29. Weathers, P. J., Dilorio, A. & Cheetham, R. D. (1989) Proceedings of Biotechnology USA Conference, San Francisco, Conference Management Corp, Norwalk, CT. pp. 247–256.

  30. Dilorio, A. A., Cheetham, R. D., & Weathers, P. J. (1992). Applied Microbiology and Biotechnology, 37, 457–462.

    Article  Google Scholar 

  31. Baveja, S. K., Rango Rao, K. V., & Arura, J. (1989). Indian Journal of Pharmaceutical Sciences, 51, 115–120.

    CAS  Google Scholar 

  32. Babu, V. S., Narasimhan, S., & Nair, G. M. (2006). Current Science, 91, 22–24.

    CAS  Google Scholar 

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Srivastava, S., Srivastava, A.K. In Vitro Azadirachtin Production by Hairy Root Cultivation of Azadirachta indica in Nutrient Mist Bioreactor. Appl Biochem Biotechnol 166, 365–378 (2012). https://doi.org/10.1007/s12010-011-9430-9

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