Biotechnology Letters

, Volume 27, Issue 5, pp 355–362 | Cite as

Domain engineering of Saccharomyces cerevisiae exoglucanases

  • S. B. Gundllapalli. Moses
  • R. R. Cordero. Otero
  • I. S. Pretorius
Article

Abstract

To illustrate the effect of a cellulose-binding domain (CBD) on the enzymatic characteristics of non-cellulolytic exoglucanases, 10 different recombinant enzymes were constructed combining the Saccharomyces cerevisiae exoglucanases, EXG1 and SSG1, with the CBD2 from the Trichoderma reesei cellobiohydrolase, CBH2, and a linker peptide. The enzymatic activity of the recombinant enzymes increased with the CBD copy number. The recombinant enzymes, CBD2-CBD2-L-EXG1 and CBD2-CBD2-SSG1, exhibited the highest cellobiohydrolase activity (17.5 and 16.3 U mg −1 respectively) on Avicel cellulose, which is approximately 1.5- to 2-fold higher than the native enzymes. The molecular organisation of CBD in these recombinant enzymes enhanced substrate affinity, molecular flexibility and synergistic activity, contributing to their elevated action on the recalcitrant substrates as characterised by adsorption, kinetics, thermostability and scanning electron microscopic analysis.

Keywords

cellulose-binding domain cellulose hydrolysis exoglucanases Saccharomyces cerevisiae 

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Copyright information

© Springer 2005

Authors and Affiliations

  • S. B. Gundllapalli. Moses
    • 1
  • R. R. Cordero. Otero
    • 1
  • I. S. Pretorius
    • 1
    • 2
  1. 1.Institute for Wine Biotechnology and Department of Viticulture & OenologyStellenbosch UniversityStellenboschSouth Africa
  2. 2.The Australian Wine Research InstituteUrrbrae, AdelaideAustralia

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