Applied Biochemistry and Biotechnology

, Volume 137, Issue 1–12, pp 711–720

Fed-Batch production of glucose 6-Phosphate dehydrogenase using recombinant saccharomyces cerevisiae

  • Luiz Carlos Martins das Neves
  • Adalberto Pessoa
  • Michele Vitolo
Session 4

Abstract

The strain Saccharomyces cerevisiae W303-181, having the plasmid YEpPGK-G6P (built by coupling the vector YEPLAC 181 with the promoter phosphoglycerate kinase 1), was cultured by fed-batch process in order to evaluate its capability in the formation of glucose 6-phosphate dehydrogenase (EC.1.1.1.49). Two liters of culture medium (10.0 g/L glucose, 3.7 g/L yeast nitrogen broth (YNB), 0.02 g/L l-tryptophan, 0.02 g/L l-histidine, 0.02 g/L uracil, and 0.02 g/L adenine) were inoculated with 1.5 g dry cell/L and left fermenting in the batch mode at pH 5.7, aeration of 2.2 vvm, 30°C, and agitation of 400 rpm. After glucose concentration in the medium was lower than 1.0 g/L, the cell culture was fed with a solution of glucose (10.0 g/L) or micronutrients (l-tryptophan, l-histidine, uracil, and adenine each one at a concentration of 0.02 g/L) following the constant, linear, or exponential mode. The volume of the culture medium in the fed-batch process was varied from 2 L up to 3 L during 5 h. The highest glucose 6-phosphate dehydrogenase activity (350 U/L; 1 U=1 µmol of NADP/min) occurred when the glucose solution was fed into the fermenter through the decreasing linear mode.

Index Entries

Fed-batch glucose 6-phosphate dehydrogenase glucose recombinant strain Saccharomyces cerevisiae fermentation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Godfrey, T. and West, S. (1996), Industrial Enzymology, 2nd ed. MacMillan, London, UK: pp. 64–65.Google Scholar
  2. 2.
    Vitolo, M. (2004), In: Enzymes as Biotechnological agents, Said, S. and Pietro, R. C. L. R. (eds.), Legis Summa, Ribeirão Preto, SP, Brazil, pp. 207–222.Google Scholar
  3. 3.
    Lojudice, F. H., Silva, D. P., Zanchiw, N. I. T., Oliveira, C. C., and Pessoa, A., Jr. (2001), Appl. Biochem. Biotechnol. 91–93, 161–169.CrossRefGoogle Scholar
  4. 4.
    Gietz, R. D. and Sugino, A. (1988), Gene 74, 527–534.CrossRefGoogle Scholar
  5. 5.
    Das Neves, L. C. M., Pessoa, A., Jr., and Vitolo, M. (2005), Biotechnol. Prog. 21, 1135–1139.CrossRefGoogle Scholar
  6. 6.
    Echegaray, O. F., Carvalho, J. C. M., Fernandes, A. N. R., Sato, S., Aquarone, E., and Vitolo, M. (2000), Biomass 19, 39–50.Google Scholar
  7. 7.
    Song, G. Y. and Chung, B. H. (1999), Process Biochem. 35, 503–508.CrossRefGoogle Scholar
  8. 8.
    Kim, C. H., Rao, K. J., Youn, D. J., and Rhee, S. K. (2003), Biotechnol. Bioprocess Eng. 8(5), 303–305.CrossRefGoogle Scholar
  9. 9.
    Miguel, A. S. M., Das Neves, L. C. M., Vitolo, M., and Pessoa, A., Jr. (2003), Biotechnol. Prog. 19, 320–324.CrossRefGoogle Scholar
  10. 10.
    Bergmeyer, H. U. (1983), In: Methods of Enzymatic Analysis, 3rd ed. Weinheim: Verlag Chemie, vol. 2, 539p.Google Scholar
  11. 11.
    Vitolo, M., Duranti, M. A., and Pellegrim, M. B. (1995), J. Ind. Microbiol. 15, 75–79.CrossRefGoogle Scholar
  12. 12.
    Brown, J. A., Segal, H. L., Maley, F., and Chu, F. (1987) J. Biol. Chem. 254, 3689–3691.Google Scholar
  13. 13.
    Schmidell, W. and Facciotti, M. C. R. (2001), In: Industrial Biotechnology, Schmidell, W., Lima, U. A. L., Aquarone, E., and Borzani, W. (eds.), Edgard Blücher, São Paulo, SP, Brazil, pp. 179–192.Google Scholar
  14. 14.
    Danesi, E. D. G., Miguel, A. S. M., Yagui, C. O. R., Carvalho, J. C. M., and Pessoa, A., Jr. (2006), J. Food Eng. 75, 96–103.CrossRefGoogle Scholar
  15. 15.
    Vitolo, M., Carvalho, J. C. M., Duranti, M. A., and Breda, M. (1991), Biomass Bioenergy 1(5), 301–304.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc 2007

Authors and Affiliations

  • Luiz Carlos Martins das Neves
    • 1
  • Adalberto Pessoa
    • 1
  • Michele Vitolo
    • 1
  1. 1.Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical SciencesUniversity of São PauloSão Paulo SPBrazil

Personalised recommendations