Advertisement

Russian Journal of Plant Physiology

, Volume 62, Issue 4, pp 557–563 | Cite as

Features of respiration and formation of steroidal glycosides in Dioscorea deltoidea cell suspension culture grown in flasks and bioreactors

  • M. V. TitovaEmail author
  • N. A. Shumilo
  • I. E. Kulichenko
  • I. M. Ivanov
  • E. S. Sukhanova
  • A. M. Nosov
Applied Aspects

Abstract

An important task is to monitor the physiological state of the cell population when scaling apparatus cultivation of higher plants’ cell cultures to industrial volumes. Investigations were carried out on the culture of Dioscorea deltoidea Wall—strain producer of steroidal glycosides (protodioscin and deltoside)—for which growth characteristics and content of furostanol glycosides were determined during the semirunning growth in bubble bioreactors (20 and 630 L); total rate of oxygen uptake and the portion of participation of alternative oxidase (AO) in the process were measured in the exponential growth phase. The measurements were performed for 3–4 subculture cycles for each apparatus, starting from the second cycle from the moment of inoculation. The values of oxygen mass transfer coefficients (K La, h−1) for bioreactors were determined at modes of mixing and aeration optimal for growing strain. The maximal portion of cyanide resistant respiration was determined in 20-liter bubble bioreactor with a point aerating device (up to 60–65% of total respiration rate). The minimal values of (K La (2.0–2.4 h−1) were obtained for the same apparatus, as well as lower levels of furostanol glycosides (4.2–8.0% of dry cell weight). Apparently, this effect was due to insufficiently uniform distribution of oxygen in the nutrient medium. Minimal activity of AO (within 15–25%) was noted for the system with more intensive mass transfer of oxygen-in 630-liter bubble bioreactor with the annular aerator (K La 7.1–8.0 h−1). Higher biosynthetic rates (7.7–13.9% of dry cell weight) were found in the same bioreactor.

Keywords

Dioscorea deltoidea plant cell suspension culture steroidal glycosides bioreactors respiration oxygen mass transfer coefficient KLa 

Abbreviations

AO

alternative oxidase

KLa

oxygen mass transfer coefficient

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Kieran, P.M., Macloughlin, P.F., and Malone, D.M., Plant cell suspension cultures: some engineering considerations, J. Biotechnol., 1997, vol. 59, pp. 39–52.PubMedCrossRefGoogle Scholar
  2. 2.
    Chattopadhyay, S., Srivastava, A.K., and Bisaria, V.S., Production of phytochemicals in plant cell bioreactors, Plant Biotechnology and Molecular Markers, Srivastava, P.S., Narula, A., and Shrivastava, S., Eds., New Delhi: Anamaya Publ., 2004, pp. 117–128.Google Scholar
  3. 3.
    Georgiev, M., Weber, J., and Maciuk, A., Bioprocessing of plant cell cultures for mass production of targeted compounds, Appl. Microbiol. Biotechnol., 2009, vol. 83, pp. 809–823.PubMedCrossRefGoogle Scholar
  4. 4.
    Strogov, S.E., Zaitseva, G.V., Belousova, I.M., Shamkov, N.V., and Simonova, G.M., Experience large-scale cultivation of Panax ginseng cells in suspension. 1. Scaling of the pilot plant, Biotekhnologya, 1990, no. 4, pp. 43–45.Google Scholar
  5. 5.
    Shamkov, N.V., Zaitseva, G.V., Belousova, I.M., Strogov, S.E., Simonova, G.M., Nosov, A.M., and Butenko, R.G., Experience large-scale cultivation of Panax ginseng cells in suspension. 2. Testing regimes for Panax ginseng cell culture on pilot plant, Biotekhnologya, 1991, no. 3, pp. 32–34.Google Scholar
  6. 6.
    Leckie, F., Scragg, A.H., and Cliffe, K.C., An investigation into the role of initial K La on the growth and alkaloid accumulation by cultures of Catharanthus roseus, Biotechnol. Bioeng., 1991, vol. 37, pp. 364–370.PubMedCrossRefGoogle Scholar
  7. 7.
    Tanaka, H., Technological problems in cultivation of plant cells at high density, Biotechnol. Bioeng., 2000, vol. 67, pp. 1203–1218.CrossRefGoogle Scholar
  8. 8.
    Marks, D.M., Equipment design considerations for large scale cell culture, Cytotechnology, 2003, vol. 42, pp. 21–33.PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Karanova, S.L., Nosov, A.M., Shamina, Z.B., and Paukov, V.N., Productivity of various cell lines of Dioscorea deltoidea, Kul’tura kletok rastenii i biotekhnologiya (Plant Cell Cultures and Biotechnology), Butenko, R.G., Ed., Moscow: Nauka, 1986, pp. 83–87.Google Scholar
  10. 10.
    Nosov, A.M., Steroid metabolism features in cultured Dioscorea deltoidea cells, as a basis for obtaining biotechnology of furostanol glycosides, Doctoral (Biol.) Dissertation, Moscow: Nauchno-issledovatel’skii i proektnokonstruktorskii institut prikladnoi biokhimii, 1992.Google Scholar
  11. 11.
    Vasil’eva, I.S. and Paseshnichenko, V.A., Steroid glycosides in plants and Dioscorea deltoidea cells, metabolism and biological activity, Usp. Biol. Khim., 2000, vol. 40, pp. 153–204.Google Scholar
  12. 12.
    Lipskii, A.Kh. and Chernyak, N.D., Effect of temperature on Dioscorea deltoidea cell culture with deep cultivation, Sov. Plant Physiol., 1983, vol. 30, pp. 437–447.Google Scholar
  13. 13.
    Kandarakov, O.F., Vorob’ev, A.S., and Nosov, A.M., Biosynthetic characteristics of Dioscorea deltoidea cell population grown in continuous culture, Russ. J. Plant Physiol., 1994, vol. 41, pp. 805–809.Google Scholar
  14. 14.
    Oreshnikov, A.V., Physiology of Dioscorea deltoidea cell culture population in a closed flow, Cand. Sci. (Biol.) Dissertation, Moscow: Timiryazev Institute Plant Physiology, Russ. Acad. Sci., 1996.Google Scholar
  15. 15.
    Murashige, T. and Skoog, F., A revised medium for rapid growth and bioassays with tobacco tissue cultures, Physiol. Plant., 1962, vol. 15, pp. 473–497.CrossRefGoogle Scholar
  16. 16.
    Kapulnik, Y., Yalpani, N., and Raskin, I., Salicylic acid induces cyanide-resistant respiration in tobacco cellsuspension cultures, Plant Physiol., 1992, vol. 100, pp. 1921–1926.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Vasil’eva, I.S., Vorob’ev, A.S., Gorskaya, N.V., Lipskii, A.Kh., Gurielidze, K.G., and Paseshnichenko, V.A., Definition of furostanol glycosides in Dioscorea deltoidea Wall. cell cultures using spectrophotometric method, Prikl. Biokhim. Mikrobiol., 1987, vol. 28, pp. 692–696.Google Scholar
  18. 18.
    Danilov, A.V., Zaitseva, G.V., Konstantinova, N.A., and Aleksandrova, I.V., Influence of intensity of mass transfer on the growth of Dioscorea deltoidea cell suspension culture, Sov. Plant Physiol., 1981, vol. 28, pp. 1072–1077.Google Scholar

Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • M. V. Titova
    • 1
    • 2
    Email author
  • N. A. Shumilo
    • 1
  • I. E. Kulichenko
    • 1
  • I. M. Ivanov
    • 1
  • E. S. Sukhanova
    • 1
    • 2
  • A. M. Nosov
    • 1
    • 2
  1. 1.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia
  2. 2.Biological FacultyMoscow State UniversityMoscowRussia

Personalised recommendations