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Applied Biochemistry and Biotechnology

, Volume 82, Issue 3, pp 243–258 | Cite as

Hydrolysis of steam-pretreated lignocellulose

Synergism and adsorption for cellobiohydrolase I and endogulcanase II of trichoderma reesei
  • Johan Karlsson
  • József Medve
  • Folke TjerneldEmail author
Article

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.

Index Entries

Trichoderma reesei cellulase cellulose hydrolysis lignocellulose cellobiohydrolase endoglucanase synergism adsorption 

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Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  1. 1.Department of Biochemistry, Center for Chemistry and Chemical EngineeringLund UniversityLundSweden

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