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The Family 1 Glycoside Hydrolase from Clostridium cellulolyticum H10 is a Cellodextrin Glucohydrolase

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Abstract

The only family 1 glycoside hydrolase in Clostridium cellulolyticum H10 (CcGH1) is annotated as a beta-galactosidase but has high sequence homology with many beta-glucosidases. Given the possible importance of beta-glucosidase in cellulose utilization by C. cellulolyticum, the encoding open reading frame Ccel_0374 was cloned and expressed in E. coli as a soluble fusion protein with thioredoxin. After tag cleavage, the purified enzyme had a molecular mass of 52 kDa and was active in dimeric form on a broad range of substrates, including cellobiose, cellotriose, cellotetraose, p-nitrophenyl-beta-glucopyranoside, lactose, and o-nitrophenyl-beta-galactopyranoside. The enzyme showed lower K m and higher catalytic efficiency (k cat/K m) on cellodextrins with degree of polymerization from 2 to 4 than on lactose, and the k cat/K m values on cellodextrins increased in the order of cellobiose < cellotriose < cellotetraose, suggesting that CcGH1 was a cellodextrin glucohydrolase (EC 3.2.1.74). The high K m (69 mM) on cellobiose implies that CcGH1 likely has a minimal role in the intracellular hydrolysis of cellobiose in C. cellulolyticum. The three-dimensional structure model of CcGH1 generated by homology modeling showed a typical (α/β)8 barrel topology characteristic of family 1 glycoside hydrolases.

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References

  1. Zhang, Y.-H. P. (2009). Energy Environment Science, 2(2), 272–282.

    Article  CAS  Google Scholar 

  2. Lynd, L. R., Weimer, P. J., van Zyl, W. H., & Pretorius, I. S. (2002). Microbiology and Molecular Biology Reviews, 66, 506–577.

    Article  CAS  Google Scholar 

  3. Zhang, Y.-H. P. (2008). Journal of Industrial Microbiology and Biotechnology, 35(5), 367–375.

    Article  CAS  Google Scholar 

  4. Ye, X., Wang, Y., Hopkins, R. C., Adams, M. W. W., Evans, B. R., Mielenz, J. R., et al. (2009). ChemSusChem, 2(2), 149–152.

    Article  CAS  Google Scholar 

  5. Atsumi, S., Hanai, T., & Liao, J. C. (2008). Nature, 451(7174), 86–89.

    Article  CAS  Google Scholar 

  6. Den Haan, R., Rose, S. H., Lynd, L. R., & van Zyl, W. H. (2007). Metabolic Engineering, 9, 87–94.

    Article  Google Scholar 

  7. Liu, W., Hong, J., Bevan, D. R., & Zhang, Y.-H. P. (2009). Biotechnology and Bioengineering, 103(6), 1087–1094.

    Article  CAS  Google Scholar 

  8. Zhang, Y.-H. P., Himmel, M., & Mielenz, J. R. (2006). Biotechnology Advances, 24(5), 452–481.

    Article  CAS  Google Scholar 

  9. Himmel, M., Tucker, M., Lastick, S., Oh, K., Fox, J., Spindler, D., et al. (1986). Journal of Biological Chemistry, 261(28), 12948–12955.

    CAS  Google Scholar 

  10. Rixon, J. E., Ferreira, L. M., Durrant, A. J., Laurie, J. I., Hazlewood, G. P., & Gilbert, H. J. (1992). Biochemical Journal, 285(3), 947–955.

    CAS  Google Scholar 

  11. Yernool, D. A., McCarthy, J. K., Eveleigh, D. E., & Bok, J.-D. (2000). Journal of Bacteriology, 182(18), 5172–5179.

    Article  CAS  Google Scholar 

  12. Desvaux, M. (2005). Enzyme and Microbial Technology, 37(4), 373–385.

    Article  CAS  Google Scholar 

  13. Doi, R. H., & Kosugi, A. (2004). Nature Reviews Microbiology, 2, 541–551.

    Article  CAS  Google Scholar 

  14. Zhang, Y.-H. P., & Lynd, L. R. (2005). Proceedings of the National Academy of Sciences of the United States of America, 102, 7321–7325.

    Article  CAS  Google Scholar 

  15. Zhang, Y.-H. P., & Lynd, L. R. (2004). Applied and Environmental Microbiology, 70, 1563–1569.

    Article  CAS  Google Scholar 

  16. Lu, Y., Zhang, Y.-H. P., & Lynd, L. R. (2006). Proceedings of the National Academy of Sciences of the United States of America, 103(44), 16165–16169.

    Article  CAS  Google Scholar 

  17. Desvaux, M. (2006). Biotechnology Progress, 22(5), 1229–1238.

    CAS  Google Scholar 

  18. Bayer, E. A., Belaich, J. P., Shoham, Y., & Lamed, R. (2004). Annual Review of Microbiology, 58, 521–554.

    Article  CAS  Google Scholar 

  19. Zhang, Y.-H. P., & Lynd, L. R. (2003). Analytical Biochemistry, 322, 225–232.

    Article  CAS  Google Scholar 

  20. van Tilbeurgh, H., Claeyssens, M., & de Bruyne, C. K. (1982). FEBS Letters, 149, 152–156.

    Article  Google Scholar 

  21. Sali, A., & Blundell, T. L. (1993). Journal of Molecular Biology, 234(3), 779–815.

    Article  CAS  Google Scholar 

  22. Cantarel, B. L., Coutinho, P. M., Rancurel, C., Bernard, T., Lombard, V., & Henrissat, B. (2009). Nucl Acids Res, 37, D233–D238.

    Article  CAS  Google Scholar 

  23. LaVallie, E. R., DiBlasio, E. A., Kovacic, S., Grant, K. L., Schendel, P. F., & McCoy, J. M. (1993). Nature Biotechnology, 11(2), 187–193.

    Article  CAS  Google Scholar 

  24. Lüthy, R., Bowie, J., & Eisenberg, D. (1992). Nature, 356, 83–85.

    Article  Google Scholar 

  25. Bhatia, Y., Mishra, S., & Bisaria, V. (2002). Critical Reviews in Biotechnology, 22, 375–407.

    Article  CAS  Google Scholar 

  26. Himmel, M. E., Adney, W. S., Baker, J. O., Nieves, R. A., & Thomas, S. R. (1996). Cellulases: structure, function and applications. In C. E. Wyman (Ed.), Handbook on bioethanol: production and utilization (pp. 144–161). Washington, DC: Taylor & Francis.

    Google Scholar 

  27. Ait, N., Creuzet, N., & Cattaneo, J. (1979). Biochemical and Biophysical Research Communications, 90(2), 537–546.

    Article  CAS  Google Scholar 

  28. Gräbnitz, F., & Staudenbauer, W. L. (1988). Biotechnological Letters, 10(2), 73–78.

    Article  Google Scholar 

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Acknowledgments

This work was supported mainly by the DOE BioEnergy Science Center and partially by the USDA Bioprocessing and Biodesign Center as well as the DuPont Young Professor Award.

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Authors and Affiliations

  1. Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, 210-A Seitz Hall, Blacksburg, VA, 24061, USA

    Wenjin Liu & Y.-H. Percival Zhang

  2. Department of Biochemistry, Virginia Polytechnic Institute and State University, 201 Fralin Hall, Blacksburg, VA, 24061, USA

    David R. Bevan

  3. Institute for Critical Technology and Applied Science (ICTAS), Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA

    Y.-H. Percival Zhang

  4. DOE BioEnergy Science Center (BESC), Oak Ridge, TN, 37831, USA

    Y.-H. Percival Zhang

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  1. Wenjin Liu
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  2. David R. Bevan
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  3. Y.-H. Percival Zhang
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Correspondence to Y.-H. Percival Zhang.

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Liu, W., Bevan, D.R. & Zhang, YH.P. The Family 1 Glycoside Hydrolase from Clostridium cellulolyticum H10 is a Cellodextrin Glucohydrolase. Appl Biochem Biotechnol 161, 264–273 (2010). https://doi.org/10.1007/s12010-009-8782-x

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  • DOI: https://doi.org/10.1007/s12010-009-8782-x

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