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Hydrolysis of steam-pretreated lignocellulose

Synergism and adsorption for cellobiohydrolase I and endogulcanase II of trichoderma reesei

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Abstract

The mechanism of hydrolysis of cellulose is important for improving the enzymatic conversion in bioprocesses based on lignocellulose. Adsorption and hydrolysis experiments were performed with cellobiohydrolase I (CBH I) and endoglucanase II (EG II) from Trichoderma reesei on a realistic lignocellulose substrates: steam-pretreated willow. The enzymes were studied both alone and in equimolar mixtures. Adsorption isotherms were determined at 4 and 40°C during 90-min reaction times. Both CBH I and EG II adsorbed stronger at 40 than at 4°C. The time course of adsorption and hydrolysis, 3 min to 48 h, was studied at 40°C. About 90% of the cellulases were adsorbed within 2 h. The hydrolysis rate was high in the beginning but decreased during the time course. Based on adsorption data, the hydrolysis and synergism were analyzed as function of adsorbed enzyme. CBH I showed a linear correlation between hydrolysis and adsorbed enzyme, whereas for EG II the corresponding curve leveled off at both 4 and 40°C. At low conversion, below 1%, EG II produced as much soluble sugars as CBH I. At higher conversion, CBH I was more efficient than EG II. The synergism as function of adsorbed enzyme increased with bound enzyme before reaching a stable value of about 2. The effect of varying the ratio of CBH I:EG II was studied at fixed total enzyme loading and by changing the ratio between the enzymes. Only a small addition (5%) of EG II to a CBH I solution was shown to be sufficient for nearly maximal synergism. The ratio between EG II and CBH I was not critical. The ratio 40% EG II:60% CBH I showed similar conversion to 5% EG II:95% CBH I. Modifications of the conventional endo-exo synergism model are proposed.

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Authors and Affiliations

  1. Department of Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, S-221 00, Lund, Sweden

    Johan Karlsson, József Medve & Folke Tjerneld

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  1. Johan Karlsson
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  2. József Medve
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  3. Folke Tjerneld
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Correspondence to Folke Tjerneld.

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Karlsson, J., Medve, J. & Tjerneld, F. Hydrolysis of steam-pretreated lignocellulose. Appl Biochem Biotechnol 82, 243–258 (1999). https://doi.org/10.1385/ABAB:82:3:243

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  • DOI: https://doi.org/10.1385/ABAB:82:3:243

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