Abstract
A critical parameter affecting the economic feasibility of lignocellulosic bioconversion is the production of inexpensive and highly active cellulase enzymes in bulk quantity. A promising approach to reduce enzyme costs is to genetically transform plants with the genes of these enzymes, thereby producing the desired cellulases in the plants themselves. Extraction and recovery of active proteins or release of active cellulase from the plants during bioconversion could have a significant positive impact on overall lignocellulose conversion economics. The effects of ammonia fiber explosion (AFEX) pretreatment variables (treatment temperature, moisture content, and ammonia loading) on the activity of plant-produced heterologous cellulase enzyme were individually investigated via heat treatmett or ammonia treatment. Finally, we studied the effects of all these variables in concert through the AFEX process. The plant materials included transgenic tobacco plants expressing E1 (endoglucanase from Acidothermus cellulolyticus). The E1 activity was measured in untreated and AFEX-treated tobacco leaves to investigate the effects of the treatment on the activity of this enzyme. The maximum observed activity retention in AFEX-treated transgenic tobacco samples compared with untreated samples was approx 35% (at 60°C, 0.5∶1 ammonia loading, and 40% moisture). Based on these findings, it is our opinion that AFEX pretreatment is not a suitable option for releasing cellulase enzyme from transgenic plants.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Wyman, C. E. (1999), Annu. Rev. Energy Environ. 24, 189–226.
Wyman, C. E. (1995), in Enzymatic Degradation of Insoluble Carbohydrates, ACS Symposium Series 618, Saddler, J. N. and Penner, M. H., eds., American Chemical Society, Washington, DC, pp. 272–290.
Lynd, L. R., Elande, R. T., and Wyman, C. E. (1996), Appl. Biochem. Biotechnol. 57/58, 741–61.
Goddijn, O. J. M. and Pen, J. (1995), Trends Biotechnol. 13, 379–387.
Kidd, G. and Devorak, J. (1994), Bio/Technology 12, 1328–1329.
Stark, D., Timmermann, K. P., Barry, G. F., Preiss, J., and Kishore, G. M. (1992), Science 258, 287–292
Grayburn, W. S., Collins, G. B., and Hildebrand, D. F. (1992), Bio/Technology 10, 675–678.
Poirier, Y., Nawrath, C., and Somerville, C. (1995), Bio/Technology 13, 142–150.
Austin, S., Koegel, R. G., Matthews, D., Shahan, M., Strau, R. J., and Burgess, R. (1994), Ann. NY Acad. Sci 721, 234–244.
Pen, J., Molendijk, L., Quax, W. J., Sijmons, P. C., van Ooyen, A. J. J., van den Elzen, P. J. M., Rietveld, K., and Hoekema, A. (1992), Bio/Technology 10, 292–296.
Pen, J., Verwoerd, T. C., van Paridon, P. A., Beudeker, R. F., van den Elzen, Geerse, K., van den Klis, J. D., Versteegh, H. A. J., van Ooyen, A. J. J., and Hoekema, A. (1993), Bio/Technology 11, 811–814.
Carroad, P. A., Anaya-Serrano, H., Edwards, R. H., and Kohler, G. O. (1981), J. Food Sci. 46(2), 383–386.
De la Rosa, L. B., Dale, B. E., Reshamwala, S. T., Latimer, V. M., Stuart, E. D., and Shawky, B. T. (1994), Appl. Biochem. Biotechnol. 45/46, 483–497.
Teymouri, F. (2003), PhD thesis, Department of Chemical Engineering and Material Sciences, Michigan State University, East Lansing, MI.
Ziegelhoffer, T., Raasch, A., and Austin-Phillips, S. (2001), Mol. Breeding 8, 147–158.
Bradford, M. (1976), Anal. Biochem. 72, 248–254.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Teymouri, F., Alizadeh, H., Laureano-Pérez, L. et al. Effects of ammonia fiber explosion treatment on activity of endoglucanase from Acidothermus cellulolyticus in transgenic plant. Appl Biochem Biotechnol 116, 1183–1191 (2004). https://doi.org/10.1385/ABAB:116:1-3:1183
Issue Date:
DOI: https://doi.org/10.1385/ABAB:116:1-3:1183