Conditions of hydrolysis with a specific pair of endo- and exo-cellulases

Enzymatic hydrolysis of cellulose is a technology involved in the production of bioethanol, a potential alternative renewable energy. Many cellulases with endo- and exo- type of activity are known to hydrolyze cellulose synergistically. In this thesis, potential synergy between an endo-cellulase, Cel5B, with and without a carbohydrate- binding module (CBM6), and a new exo-cellulase, CBH1, from Trichoderma harzianum FP108 were examined during the hydrolysis of semi- crystalline cellulose (Avicel). Since CBM6 is recognized as having a high affinity for amorphous cellulose, it was hypothesized that this affinity could enhance the synergistic effect between the endo- and exo-cellulases by focusing the action to Cel5B+CBM6 on the amorphous regions of the Avicel substrate. The increased activity of Cel5B+CBM6 over Cel5B alone was confirmed. However, in contrast to our expectations, a synergistic effect was not observed between either endo- and exo-cellulase pairs. From the obtained hydrolysis yield, it was inferred that Cel5B+CBM6 may have exo-type activity that caused a competitive interaction with the exo-cellulase, which resulted in no synergy.