Applied Biochemistry and Biotechnology

, Volume 163, Issue 4, pp 485–496 | Cite as

Adaptive Evolution of Escherichia coli Inactivated in the Phosphotransferase System Operon Improves Co-utilization of Xylose and Glucose Under Anaerobic Conditions

  • Victor Emmanuel Balderas-Hernández
  • Verónica Hernández-Montalvo
  • Francisco Bolívar
  • Guillermo Gosset
  • Alfredo Martínez


Modification of the phosphoenolpyruvate/sugar phosphotransferase system (PTS) has shown improvement in sugar coassimilation in Escherichia coli production strains. However, in preliminary experiments under anaerobic conditions, E. coli strains with an inactive PTS and carrying pLOI1594, which encodes pyruvate decarboxylase and alcohol dehydrogenase from Zymomonas mobilis, were unable to grow. These PTS strains were previously evolved under aerobic conditions to grow rapidly in glucose (PTS- Glucose+ phenotype). Thus, in this work, applying a continuous culture strategy under anaerobic conditions, we generate a new set of evolved PTS Glucose+ mutants, VH30N1 to VH30N6. Contrary to aerobically evolved mutants, strains VH30N2 and VH30N4 carrying pLOI1594 grew in anaerobiosis; also, their growth capacity was restored in a 100%, showing specific growth rates (μ ~ 0.12 h−1) similar to the PTS+ parental strain (μ = 0.11 h−1). In cultures of VH30N2/pLOI1594 and VH30N4/pLOI1594 using a glucose–xylose mixture, xylose was totally consumed and consumption of sugars occurred in a simultaneous manner indicating that catabolic repression is alleviated in these strains. Also, the efficient sugar coassimilation by the evolved strains caused an increment in the ethanol yields.


Anaerobiosis Evolved strains PTS Sugar co-utilization Xylose 



We thank Mercedes Enzaldo, Georgina Hernández, and Roberto Rodríguez for technical assistance. This work was supported by the following grants: Estado de Morelos MOR-2007-C01-80360 and PAPIIT-DGAPA-UNAM IN226908-3 and IN220908. V. Hernández-Montalvo held a scholarship from CONACyT-México. V.E. Balderas-Hernández is thankful for the postdoctoral fellowship support by DGAPA-UNAM.


  1. 1.
    Shanmugam, K. T., & Ingram, L. O. (2008). Journal of Molecular Microbiology and Biotechnology, 15, 8–15.CrossRefGoogle Scholar
  2. 2.
    Wang, Z., Chen, M., Xu, Y., Li, S., Lu, W., Ping, S., et al. (2008). Biotechnology Letters, 30, 657–663.CrossRefGoogle Scholar
  3. 3.
    Lin, Y., & Tanaka, S. (2006). Applied Microbiology and Biotechnology, 69, 627–642.CrossRefGoogle Scholar
  4. 4.
    Dien, B. S., Cotta, M. A., & Jeffries, T. W. (2003). Applied Microbiology and Biotechnology, 63, 258–266.CrossRefGoogle Scholar
  5. 5.
    Görke, B., & Stülke, J. (2008). Nature Reviews. Microbiology, 6, 613–624.CrossRefGoogle Scholar
  6. 6.
    Deutscher, J. (2008). Current Opinion in Microbiology, 11, 87–93.CrossRefGoogle Scholar
  7. 7.
    Saier, M. H. (1977). Bacteriological Reviews, 41, 856–871.Google Scholar
  8. 8.
    Kotrba, P., Inui, M., & Yukawa, H. (2001). Journal of Bioscience and Bioengineering, 92, 502–517.CrossRefGoogle Scholar
  9. 9.
    Flores, N., Xiao, J., Berry, A., Bolivar, F., & Valle, F. (1996). Nature, 4, 620–623.Google Scholar
  10. 10.
    Flores, S., Gosset, G., Flores, N., de Graff, A. A., & Bolivar, F. (2002). Metabolic Engineering, 4, 124–137.CrossRefGoogle Scholar
  11. 11.
    Flores, N., Flores, S., Escalante, A., de Anda, R., Leal, L., Malpica, R., et al. (2005). Metabolic Engineering, 7, 70–87.CrossRefGoogle Scholar
  12. 12.
    Flores, S., Flores, N., de Anda, R., González, A., Escalante, A., Sigala, J. C., et al. (2005). Journal of Molecular Biology and Biotechnology, 10, 51–63.Google Scholar
  13. 13.
    Hernández-Montalvo, V., Valle, F., Bolívar, F., & Gosset, G. (2001). Applied Microbiology and Biotechnology, 57, 186–191.CrossRefGoogle Scholar
  14. 14.
    Hernández-Montalvo, V., Martínez, A., Hernández-Chávez, G., Bolívar, F., Valle, F., & Gosset, G. (2003). Biotechnology and Bioengineering, 83, 687–694.CrossRefGoogle Scholar
  15. 15.
    Maniatis, T., Fritsch, E. F., & Sambrook, J. (1989). In Nolan C (ed), Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor: Cold Spring Harbor Laboratory.Google Scholar
  16. 16.
    Jensen, K. F. (1993). Journal of Bacteriology, 175, 3401–3407.Google Scholar
  17. 17.
    Martinez, A., York, S. W., Yomano, L. P., Pineda, V. L., Davis, F. C., Shelton, J. C., et al. (1999). Biotechnology Progress, 15, 891–897.CrossRefGoogle Scholar
  18. 18.
    Beall, D. S., Ohta, K., & Ingram, L. O. (1991). Biotechnology and Bioengineering, 38, 296–303.CrossRefGoogle Scholar
  19. 19.
    Martinez, A., Rodriguez, M. E., Wells, M. L., York, S. W., Preston, J. F., & Ingram, L. O. (2001). Biotechnology Progress, 17, 287–293.CrossRefGoogle Scholar
  20. 20.
    Madhavan, A., Tamalampudi, S., Srivastava, A., Fukuda, H., Bisaria, V. S., & Kondo, A. (2009). Applied Microbiology and Biotechnology, 82, 1037–1047.CrossRefGoogle Scholar
  21. 21.
    Hasona, A., Kim, Y., Healy, F. G., Ingram, L. O., & Shanmugam, K. T. (2004). Journal of Bacteriology, 186, 7593–7600.CrossRefGoogle Scholar
  22. 22.
    Okuda, N., Ninomiya, K., Takao, M., Katakura, Y., & Shioya, S. (2007). Journal of Bioscience and Bioengineering, 103, 350–357.CrossRefGoogle Scholar
  23. 23.
    Dien, B. S., Nichols, N. N., & Bothast, R. J. (2002). Journal of Industrial Microbiology and Biotechnology, 29, 221–227.CrossRefGoogle Scholar
  24. 24.
    Nichols, N. N., Dien, B. S., & Bothast, R. J. (2001). Applied Microbiology and Biotechnology, 56, 120–125.CrossRefGoogle Scholar
  25. 25.
    Cirino, P. C., Chin, J. W., & Ingram, L. O. (2006). Biotechnology and Bioengineering, 95, 1167–1176.CrossRefGoogle Scholar
  26. 26.
    Khankal, R., Chin, J. W., & Cirino, P. C. (2008). Journal of Biotechnology, 134, 246–252.CrossRefGoogle Scholar
  27. 27.
    Eiteman, M. A., Lee, S. A., & Altman, E. (2008). Journal of Biological Engineering, 2, 3.CrossRefGoogle Scholar
  28. 28.
    Andersson, C., Hodge, D., Berglund, K. A., & Rova, U. (2007). Biotechnology Progress, 23, 381–388.CrossRefGoogle Scholar
  29. 29.
    Li, R., Chen, Q., Wang, P. G., & Qi, Q. (2007). Applied Microbiology and Biotechnology, 75, 1103–1109.CrossRefGoogle Scholar
  30. 30.
    Mascarenhas, D. (1987). Patent WO/1987/001130, US.Google Scholar
  31. 31.
    Escalante, A., Calderón, R., Valdivia, A., de Anda, R., Hernández, G., Ramírez, O.T., et al. (2010). Microbial Cell Factories, 9:21, 1–12.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Victor Emmanuel Balderas-Hernández
    • 1
  • Verónica Hernández-Montalvo
    • 1
  • Francisco Bolívar
    • 1
  • Guillermo Gosset
    • 1
  • Alfredo Martínez
    • 1
  1. 1.Departamento de Ingeniería Celular y Biocatálisis, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMéxico

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