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Purification and Characterization of Recombinant Cel7A from Maize Seed

Applied Biochemistry and Biotechnology volume 174, pages 2864–2874 (2014)Cite this article

Abstract

The corn grain biofactory was used to produce Cel7A, an exo-cellulase (cellobiohydrolase I) from Hypocrea jecorina. The enzymatic activity on small molecule substrates was equivalent to its fungal counterpart. The corn grain-derived enzyme is glycosylated and 6 kDa smaller than the native fungal protein, likely due to more sugars added in the glycosylation of the fungal enzyme. Our data suggest that corn seed-derived cellobiohydrolase (CBH) I performs as well as or better than its fungal counterpart in releasing sugars from complex substrates such as pre-treated corn stover or wood. This recombinant protein product can enter and expand current reagent enzyme markets as well as create new markets in textile or pulp processing. The purified protein is now available commercially.

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References

  1. Schulein, M. (2000). Biochimica et Biophysica Acta, 1543, 239–252.

    CAS  Article  Google Scholar 

  2. Tomme, P., Warren, R. A., & Gilkes, N. R. (1995). Advances in Microbial Physiology, 37, 1–81.

    CAS  Article  Google Scholar 

  3. Hood, E. E., Love, R., Lane, J., Bray, J., Clough, R., Pappu, K., Drees, C., Hood, K. R., Yoon, S., Ahmad, A., & Howard, J. A. (2007). Plant Biotechnology Journal, 5, 709–719.

    CAS  Article  Google Scholar 

  4. Hood, E. E., Devaiah, S. P., Fake, G., Egelkrout, E., Teoh, K., Requesens, D. V., Hayden, C., Hood, K. R., Pappu, K. M., Carroll, J., & Howard, J. A. (2012). Plant Biotechnology Journal, 10, 20–30.

    CAS  Article  Google Scholar 

  5. Hood, E. E., Bailey, M. R., Beifuss, K., Magallanes-Lundback, M., Horn, M. E., Callaway, E., Drees, C., Delaney, D. E., Clough, R., & Howard, J. A. (2003). Plant Biotechnology Journal, 1, 129–140.

    CAS  Article  Google Scholar 

  6. Hood, E. E., & Woodard, S. (2002). Industrial proteins produced from plants. In E. E. Hood & J. A. Howard (Eds.), Plants as factories for protein production (pp. 119–135). The Netherlands: Kluwer Academic Publishers Dordrecht.

    Chapter  Google Scholar 

  7. Woodard, S. L., Mayor, J. M., Bailey, M. R., Barker, D. K., Love, R. T., Lane, J. R., Delaney, D. E., McComas-Wagner, J. M., Mallubhotla, H. D., Hood, E. E., Dangott, L. J., Tichy, S. E., & Howard, J. A. (2003). Biotechnology and Applied Biochemistry, 38, 123–130.

    CAS  Article  Google Scholar 

  8. Ziegler, M., Thomas, S. R., & Danna, K. (2000). Molecular Breeding, 6, 37–46.

    CAS  Article  Google Scholar 

  9. Ziegelhoffer, T., Will, J., & Austin-Phillips, S. (1999). Molecular Breeding, 5, 309–318.

    CAS  Article  Google Scholar 

  10. Dai, Z., Hooker, B., Anderson, D. B., & Thomas, S. R. (2000). Transgenic Research, 9, 43–54.

    CAS  Article  Google Scholar 

  11. Howard, J., Nikolov, Z., & Hood, E. (2011). Enzyme production systems for biomass conversion. In E. Hood, P. Nelson, & R. Powell (Eds.), Plant biomass conversion (pp. 227–253). Ames: Wiley-Blackwell.

    Chapter  Google Scholar 

  12. Walpole, G. S. (1914). CCXXXIII. Journal of the Chemical Society, Transactions, 105, 2501–2521.

    CAS  Article  Google Scholar 

  13. Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.

    CAS  Article  Google Scholar 

  14. Buchanan, B. B., Gruissem, W. and Jones, R. L. (2002) Biochemistry & Molecular Biology of Plants. American Society of Plant Biologists, Rockville, MD

  15. Jeoh, T., Michener, W., Himmel, M. E., Decker, S. R., & Adney, W. S. (2008). Biotechnology for Biofuels, 1, 10–22.

    Article  Google Scholar 

  16. Adney, W., S., Chou, Y.-C., Decker Stephen, R., Ding, S.-Y., Baker John, O., Kunkel, G., Vinzant Todd, B. and Himmel Michael, E. (2003) Heterologous expression of Trichoderma reesei 1,4-beta-D-glucan cellobiohydrolase (Cel 7A). In Applications of enzymes to lignocellulosics (pp. 403–437). Washington: American Chemical Society

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Acknowledgments

This study was supported by a Phase 1 Small Business Innovation Research Grant from the National Institute for Food and Agriculture (ARKW-2011-00108).

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Authors and Affiliations

  1. Infinite Enzymes, LLC, PO Box 2654, State University, AR, 72467, USA

    Nathan C. Hood, Kendall R. Hood & Elizabeth E. Hood

  2. Kalon Biotherapeutics, 100 Discovery Drive, Suite 200, College Station, TX, 77845, USA

    Susan L. Woodard

  3. Department of Biological Sciences, East Tennessee State University, Johnson City, TN, 37614, USA

    Shivakumar P. Devaiah

  4. Department of Biological and Agricultural Engineering, University of California, Davis, CA, 95616, USA

    Tina Jeoh

  5. Biological and Agricultural Engineering, Kansas State University, Manhattan, KS, 66506-2906, USA

    Lisa Wilken

  6. Biological and Agricultural Engineering, Texas A&M University, College Station, TX, 77843, USA

    Zivko Nikolov

  7. Applied Biotechnology Institute, Cal Poly Tech Park, San Luis Obispo, CA, 93407, USA

    Erin Egelkrout & John A. Howard

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  1. Nathan C. Hood
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  2. Kendall R. Hood
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  3. Susan L. Woodard
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  4. Shivakumar P. Devaiah
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  5. Tina Jeoh
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  6. Lisa Wilken
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  7. Zivko Nikolov
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  8. Erin Egelkrout
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  9. John A. Howard
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  10. Elizabeth E. Hood
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Correspondence to Elizabeth E. Hood.

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Hood, N.C., Hood, K.R., Woodard, S.L. et al. Purification and Characterization of Recombinant Cel7A from Maize Seed. Appl Biochem Biotechnol 174, 2864–2874 (2014). https://doi.org/10.1007/s12010-014-1232-4

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  • DOI: https://doi.org/10.1007/s12010-014-1232-4

Keywords

  • Cel7A
  • Recombinant protein
  • Cellobiohydrolase I
  • Protein purification
  • Cellulase
  • Biomass conversion