Skip to main content
Log in

Effects of glycerol concentration and pH on growth of recombinantPichia pastoris yeast

  • Original Ariticle
  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Batch fermentations were used to study the effect of different glycerol concentrations and pH conditions on growth of recombinantPichia pastoris. Two strains ofP. pastoris were used: a wild-type in methanol utilization (Mut+) and a mutant defective in methanol utilization (Mut-). Under constant pH conditions of 5.0, glycerol concentrations up to 12% were efficiently utilized. Cell yield (Yx/s) of about 0.8 and a final cell density of about 95 g/L (dry cell) were achieved. However, there were significant differences (probability [Pr]> F 0.0351) in specific growth rates between the initial glycerol concentrations of 2, 7, and 12%. When fermentations were conducted without pH control, growth continued until the pH had decreased to about 2.5. Growth stopped at pH 2.2 with uncontrolled pH, and residual glycerol concentrations were greater than 2%. As a result, Yx/s decreased to about 0.3. There were no differences between Mut+ and Mut- strains during cell growth on glycerol.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Cregg, J. M., Vedvick, T. S., and Raschke, W. C. (1993),Bio/Technology 11, 905–910.

    Article  CAS  Google Scholar 

  2. Sreekrishna, K., Nelles, L., Potenz, R., Cruze, J., Mazzaferro, P., Fish, W., et al. (1989),Biochemistry 28, 4117–4125.

    Article  CAS  Google Scholar 

  3. Tschopp, J. F., Sverlow, G., Kosson, R., Craig, W., and Grinna, L. (1987),Bio/Technology 5, 1305–1308.

    Article  CAS  Google Scholar 

  4. Hitzman, D. L., Wegner, G. H., and Shay, L. K. (1987), inBiological Research on Industrial Yeast, vol. 1 (Stewart, G. G., Russell, I., Klein, R. D., and Heibsch, R. R., eds.), CRC, Boca Raton, FL, pp. 21–25.

    Google Scholar 

  5. Couderc, R. and Baratti, J. (1980),Agric. Biol. Chem. 44, 2279–2289.

    CAS  Google Scholar 

  6. Brierley, R. A., Bussineau, C., Kosson, R., Melton, A., and Siegel, R. S. (1990),Ann. NY Acad. Sci. 6, 350–362.

    Article  Google Scholar 

  7. Bech-Jensen E. and Carlsen, S. (1990),Biotechnol. Bioeng. 36, 1–11.

    Article  Google Scholar 

  8. Barford, J. P. and Hall, R. J. (1979),J. Gen. Microbiol. 114, 267–275.

    CAS  Google Scholar 

  9. Doelle, H. W., Ewings, K. N., and Hollywood, N. W. (1982),Adv. Biochem. Eng/Biotech. 23, 1–35.

    CAS  Google Scholar 

  10. Barnett, J. A. (1976), inAdvances in Carbohydrate Chemistry and Biochemistry, vol. 32 (Tipson, R. S. and Horton, D., eds.), Academic, New York, pp. 125–234.

    Google Scholar 

  11. Gancedo, C., Gancedo, J. M., and Sols, A. (1968),Eur. J. Biochem. 5, 165–172.

    Article  CAS  Google Scholar 

  12. Sprague, G. F. and Cronan, J. E. (1977),J. Bacteriol. 129, 1335–1342.

    CAS  Google Scholar 

  13. Clare, J. J., Romanos, M. A., Rayment, F. B., Rowedder, J. E., Smith, M. A., Payne, M. M, Sreekrishna, K., and Henwood, C. A. (1991),Gene 105, 205–212.

    Article  CAS  Google Scholar 

  14. Siegel, R. S., Buckholz, R. G., Thill, G. P., and Wondrack, L. M. (1990), International Patent Application, Publication No. WO90/10697.

  15. Scorer, C. A., Buckholz, R. G., Clare, J. J., and Romanos, M. A. (1993),Gene 136, 111–119.

    Article  CAS  Google Scholar 

  16. Cregg, J. M., Madden, K. R., Barringer, K. J., Thill, G. P., and Stillman, C. A. (1989),Mol. Cell. Biol. 9, 1316–1323.

    CAS  Google Scholar 

  17. Cregg, J. M. and Madden, K. R. (1988),Dev. Ind. Microbiol. 29, 33–41

    CAS  Google Scholar 

  18. Chiruvolu, V., Cregg, J. M., and Meagher, M. M. (1997),Enzyme Microb. Technol. 21, 227–283.

    Article  Google Scholar 

  19. Becker, J. U., Vohmann, H. J., and Eilers-Konig, C. (1979),Arch. Microbiol. 123, 143–149.

    Article  CAS  Google Scholar 

  20. Panek, A. D. (1991), inThe Yeasts (Rose, A. H. and Harrison, J. S., eds.), Academic, New York, pp. 655–678.

    Google Scholar 

  21. Werner-Washburne, M., Braun, E., Johnston, G. C., and Singer, R. A. (1993),Microbiol. Rev. 6, 383–401.

    Google Scholar 

  22. Dombek, K. M. and Ingram, L. O. (1987),Appl. Environ. Microbiol. 53, 1286–1291.

    CAS  Google Scholar 

  23. Imai, T. and Ohno, T. (1995)J. Biotechnol. 38, 165–172.

    Article  CAS  Google Scholar 

  24. Veenhuis, J. J., VanDijken, P., and Harder, W. (1983), inAdvances in Microbial Physiology, vol. 24 Academic, New York, pp. 1–82.

    Google Scholar 

  25. Tanner, R. D., Souki, N. T., and Russell R. M. (1977),Biotechnol. Bioeng. 19, 27–42.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This article is published as paper No. 11607, Journal Series, Nebraska Agricultural Experiment Station, Lincoln, NE 68583-0704.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chiruvolu, V., Eskridge, K., Cregg, J. et al. Effects of glycerol concentration and pH on growth of recombinantPichia pastoris yeast. Appl Biochem Biotechnol 75, 163–173 (1998). https://doi.org/10.1007/BF02787771

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02787771

Index entries

Navigation