Original Research Paper

Biotechnology Letters

, Volume 34, Issue 1, pp 91-96

Trichoderma reesei cellobiohydrolase II is associated with the outer membrane when overexpressed in Escherichia coli

  • Diya M. AbdeljabbarAffiliated withDepartment of Chemical & Biological Engineering, Princeton University
  • , Hank J. SongAffiliated withDepartment of Chemical & Biological Engineering, Princeton University
  • , A. James LinkAffiliated withDepartment of Chemical & Biological Engineering, Princeton UniversityDepartment of Molecular Biology, Princeton UniversityA207 Engineering Quadrangle, Princeton University Email author 

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Abstract

Cellulose degradation is essential for the future production of many advanced biofuels. Cellulases from the filamentous fungus Trichoderma reesei are among the most efficient enzymes for the hydrolysis of cellulosic materials. One of the cellulases from T. reesei, cellobiohydrolase II (CBH2), was studied because of its industrial relevance and proven enzymatic activity. Using both crude and rigorous membrane fractionation methods we show that full length T. reesei CBH2 is exclusively localized to the outer membrane when expressed recombinantly in Escherichia coli. Even fusing signal sequence-free maltose-binding protein to the N-terminus of CBH2, which has been shown to increase solubility of other proteins, did not prevent the outer membrane localization of CBH2. These results highlight the difficulties in producing fungal cellulases in bacterial hosts and provide a stepping stone for future cellulase engineering efforts.

Keywords

Fungal cellulase Fusion protein Heterologous protein expression Membrane localization