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Enhanced Expression Levels of Cellulase Enzymes Using Multiple Transcription Units

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Abstract

Transgenic cereals are an attractive option for the accumulation of foreign proteins when large volumes and low cost are required. Previous work has shown maize germ to be a particularly good location for accumulating enzymes that target cellulose for degradation. In this study, recently identified embryo-preferred promoters were used to investigate their ability to increase the accumulation of the enzymes endoglucanase E1 and cellobiohydrolase CBHI. The effect of increasing copy numbers of identical transcription units, as well as multiple copies of the enzyme driven by different promoters, was explored. Results show that accumulation of the E1 or CBHI enzymes can be significantly increased, particularly when using constructs with multiple copies of the transcription units. These findings demonstrate the highest levels of these enzymes obtained in a commercially relevant plant species observed thus far. The methodology described here may provide a low-cost plant-based source of enzymes enabling an economically viable solution for the conversion of cellulose to ethanol.

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Acknowledgments

This work was supported by grant #DOE DE FG36 GO88025, Foundation, Walton Family Foundation, and Arkansas State University Biosciences Institute. We would also like to acknowledge the technical assistance of Aaron Harry, Mackenzie Tageson, and Raghavendra Rayadurg in this project.

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Authors and Affiliations

  1. Applied Biotechnology Institute, Cal Poly State University, Building 83, Suite 1D, San Luis Obispo, CA, 93407, USA

    Erin Egelkrout, Todd Keener, Amberlyn Ferleman & John Howard

  2. Department of Statistics, Cal Poly State University, San Luis Obispo, CA, 93407, USA

    Karen McGaughey

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

    Susan Woodard

  4. Arkansas Biosciences Institute, Arkansas State University, Jonesboro, AR, 72467, USA

    Shivakumar Devaiah & Elizabeth Hood

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

    Zivko Nikolov

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  1. Erin Egelkrout
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  2. Karen McGaughey
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  3. Todd Keener
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  4. Amberlyn Ferleman
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  8. Elizabeth Hood
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  9. John Howard
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Correspondence to John Howard.

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Egelkrout, E., McGaughey, K., Keener, T. et al. Enhanced Expression Levels of Cellulase Enzymes Using Multiple Transcription Units. Bioenerg. Res. 6, 699–710 (2013). https://doi.org/10.1007/s12155-012-9288-x

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  • DOI: https://doi.org/10.1007/s12155-012-9288-x

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