Skip to main content
SpringerLink
Log in

Protein Engineering of the Soluble Metal-dependent Formate Dehydrogenase from Escherichia coli

  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

Formate is the most targeted C1 building block and electron carrier in the post-petroleum era. Formate dehydrogenase (FDH), which catalyzes the production or degradation of formate, has acquired considerable attention. Among FDHs, a metal-dependent FDH that carries a complex active center, molybdenum-pterin cofactor, can directly transfer electrons from formate to other redox proteins without generating NAD(P)H. Previously, we reported an expression system for membrane-bound metal-dependent FDH from E. coli (encoded by the fdoG-fdoH-fdoI operon) and succeeded in its conversion to a soluble protein. However, this protein exhibited a too low stability to be purified and analyzed biochemically. In this study, we tried to improve the stability of heterologously expressed FDH through rational and irrational approaches. As a result, a mutant with the highest specific activity was obtained through a rational approach. This study not only yielded a promising FDH enzyme with enhanced activity and stability for industrial applications, but also offered relevant insights for the handling of recombinant large proteins.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. G. Centi, E. A. Quadrelli, and S. Perathoner, Energ. Environ. Sci., 2013, 6, 1711.

    Article  CAS  Google Scholar 

  2. E. A. Quadrelli, G. Centi, J. L. Duplan, and S. Perathoner, ChemSusChem, 2011, 4, 1194.

    Article  CAS  PubMed  Google Scholar 

  3. IRENA, International Renewable Energy Agency, Abu Dhabi, https://www.irena.org/-Zmedia/Files/IRENA/Agency/ Publication/2019/Sep/IRENA_Hydrogen_2019.pdf, 2019.

  4. J. Eppinger and K. W. Huang, ACS Energy Lett., 2017, 2, 188.

    Article  CAS  Google Scholar 

  5. O. Aldosari, Turk. J. Chem, 2019, 43, 995.

    Article  CAS  Google Scholar 

  6. Q. Liu, X. F Yang, L. Li, S. Miao, Y. Li, Y. Q. Li, X. K. Wang, Y. Q. Huang, and T. Zhang, Nat. Commun., 2017, 8.

  7. T. G. Ostapowicz, M. Schmitz, M. Krystof, J. Klankermayer, and W. Leitner, Angew. Chem. Int. Edit., 2013, 52, 12119.

    Article  CAS  Google Scholar 

  8. M. Ihara, Y. Kawano, Y. Fujiwara, T. Kodo, M. Mizuguchi, Y. Mochiduki, K. Kodoh, A. Okabe, and I. Matsuno, J. Photochem. Photobiol. A, 2015, 313, 154.

    Article  CAS  Google Scholar 

  9. M. Ihara, H. Nakamoto, T. Kamachi, I. Okura, and M. Maeda, Photochem. Photobiol., 2006, 82, 1677.

    Article  CAS  PubMed  Google Scholar 

  10. M. Ihara, H. Nishihara, K. S. Yoon, O. Lenz, B. Friedrich, H. Nakamoto, K. Kojima, D. Honma, T. Kamachi, and I. Okura, Photochem. Photobiol., 2006, 82, 676.

    Article  CAS  PubMed  Google Scholar 

  11. M. Ihara, Y. Kawano, M. Urano, and A. Okabe, PLoS One, 2013, 8, e71581.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. C. H. Chang and K. Kim, J. Chem. Theory Comput., 2009, 5, 1137.

    Article  CAS  PubMed  Google Scholar 

  13. M. Xue, T. Wakamoto, C. Kejlberg, Y. Yoshimura, T. A. Nielsen, M. W. Rispr, K. W. Sanggaard, R. Kitahara, and F. A. A. Mulder, Proc. Natl. Acad. Sci. U. S. A., 2019, 116, 21031.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. V. G. Eijsink, B. W. Dijkstra, G. Vriend, J. R. van der Zee, O. R. Veltman, B. van der Vinne, B. van den Burg, S. Kempe, and G. Venema, Protein Eng., 1992, 5, 421.

    Article  CAS  PubMed  Google Scholar 

  15. K. Ishikawa, H. Nakamura, K. Morikawa, and S. Kanaya, Biochemistry, 1993, 32, 6171.

    Article  CAS  PubMed  Google Scholar 

  16. H. Kono, M. Saito, and A. Sarai, Proteins, 2000, 38, 197.

    Article  CAS  PubMed  Google Scholar 

  17. T. Ohmura, T. Ueda, K. Ootsuka, M. Saito, and T. Imoto, Protein Sci., 2001, 10, 313.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. T. Sengupta, Y. Tsutsui, and P. L. Wintrode, Biochemistry, 2009, 48, 8233.

    Article  CAS  PubMed  Google Scholar 

  19. R. Sant’Anna, M. R. Almeida, N. Varejao, P. Gallego, S. Esperante, P. Ferreira, A. Pereira-Henriques, F. L. Palhano, M. de Carvalho, D. Foguel, D. Reverter, M. J. Saraiva, and S. Ventura, Sci. Rep., 2017, 7, 44709.

    Article  PubMed  PubMed Central  Google Scholar 

  20. S. M. da Silva, C. Pimentel, F. M. Valente, C. Rodrigues- Pousada, and I. A. Pereira, J. Bacteriol., 2011, 193, 2909.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. N. Schwanhold, C. Iobbi-Nivol, A. Lehmann, and S. Leimkuhler, PLoS One, 2018, 13, e0201935.

    Article  PubMed  PubMed Central  Google Scholar 

  22. C. M. English, C. Eckert, K. Brown, M. Seibert, and P. W. King, Dalton Trans., 2009, 9970.

  23. J. Sun, R. C. Hopkins, F. E. Jenney, P. M. McTernan, and M. W. Adams, PLoS One, 2010, 5, e10526.

    Article  PubMed  PubMed Central  Google Scholar 

  24. J. Schiffels, O. Pinkenburg, M. Schelden, E. H. A. A. Aboulnaga, M. E. M. Baumann, and T. Selmer, Plos One, 2013, 8, e68812.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. C. H. Wu, P. M. McTernan, M. E. Walter, and M. W. Adams, Archaea, 2015, 2015, 912582.

    Article  PubMed  PubMed Central  Google Scholar 

  26. C. H. Wu, C. A. Ponir, D. K. Haja, and M. W. W. Adams, Protein Eng. Des. Sel., 2018, 31, 337.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Bioscience and Food Production Science, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 8304 Minami-minowa, Kamiina, Nagano, 399-4598, Japan

    Rintaro Fuji, Koji Umezawa, Manami Mizuguchi & Masaki Ihara

  2. Institutes for Biomedical Sciences Interdisciplinary Cluster for Cutting Edge Research, Shinshu University, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan

    Koji Umezawa & Masaki Ihara

Authors
  1. Rintaro Fuji
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Koji Umezawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manami Mizuguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Masaki Ihara
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Masaki Ihara.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fuji, R., Umezawa, K., Mizuguchi, M. et al. Protein Engineering of the Soluble Metal-dependent Formate Dehydrogenase from Escherichia coli. ANAL. SCI. 37, 733–739 (2021). https://doi.org/10.2116/analsci.20SCP15

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2116/analsci.20SCP15

Keywords

Search

Navigation