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Directed Evolution of a Fungal Xylanase for Improvement of Thermal and Alkaline Stability

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Laboratory Protocols in Fungal Biology

Part of the book series: Fungal Biology ((FUNGBIO))

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Abstract

Pre-treatment of paper pulps with xylanases has been shown to decrease the amounts of toxic chlorine dioxide used to bleach pulp. Natural xylanases are unable to tolerate the extremes of pH and temperature during the paper bleaching process and have to be genetically modified to be made more suitable for such industrial conditions. Such modification can be achieved using site-directed or random mutagenesis methods. Random mutagenesis methods are more attractive because detailed information regarding sequence or structure of the enzyme is not required. This chapter outlines how the thermal stability and alkaline stability of a glycosyl hydrolase family 11 cellulase-free xylanase from the fungus Thermomyces lanuginosus were improved using two random mutagenesis methods, error-prone PCR and a DNA shuffling method called the staggered extension process.

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Acknowledgements

Ms. Siphi Dlungwane is duly acknowledged for providing technical assistance. The National Research Foundation of South Africa funded this research.

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Authors and Affiliations

  1. Department of Biotechnology and Food Technology, ML Sultan Campus, Durban University of Technology, Durban, Kwa-Zulu-Natal, 4001, South Africa

    Dawn Elizabeth Stephens, Suren Singh & Kugen Permaul

Authors
  1. Dawn Elizabeth Stephens
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  2. Suren Singh
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  3. Kugen Permaul
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Corresponding author

Correspondence to Dawn Elizabeth Stephens .

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Editors and Affiliations

  1. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Vijai Kumar Gupta

  2. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Maria G. Tuohy

  3. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Manimaran Ayyachamy

  4. , Manufacturing Sciences and Technology, Pfizer Ireland Pharmaceuticals, The Pfizer Biotech Campus at Grange Castle, Clondalkin, N/A, Dublin, Ireland

    Kevin M. Turner

  5. , School of Natural Sciences, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Anthonia O’Donovan

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Stephens, D.E., Singh, S., Permaul, K. (2013). Directed Evolution of a Fungal Xylanase for Improvement of Thermal and Alkaline Stability. In: Gupta, V., Tuohy, M., Ayyachamy, M., Turner, K., O’Donovan, A. (eds) Laboratory Protocols in Fungal Biology. Fungal Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-2356-0_40

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