title: Adsorption of Trichoderma reesei CBH I and EG II and their catalytic domains on steam pretreated softwood and isolated lignin.
abstract: The presence of lignin has shown to play an important role in the enzymatic degradation of softwood. The adsorption of enzymes, and their constituent functional domains on the lignocellulosic material is of key importance to fundamental knowledge of enzymatic hydrolysis. In this study, we compared the adsorption of two purified cellulases from Trichoderma reesei, CBH I (Cel7A) and EG II (Cel5A) and their catalytic domains on steam pretreated softwood (SPS) and lignin using tritium labeled enzymes. Both CBH I and its catalytic domain exhibited a higher affinity to SPS than EG II or its catalytic domain. Removal of cellulose binding domain decreased markedly the binding efficiency. Significant amounts of CBH I and EG II also bound to isolated lignin. Surprisingly, the catalytic domains of the two enzymes of T. reesei differed essentially in the adsorption to isolated lignin. The catalytic domain of EG II was able to adsorb to alkaline isolated lignin with a high affinity, whereas the catalytic domain of CBH I did not adsorb to any of the lignins tested. The results indicate that the cellulose binding domain has a significant role in the unspecific binding of cellulases to lignin.
authors.affiliation: VTT Biotechnology, P.O. Box 1500, FIN-02044 VTT, Espoo, Finland. Hetti.Palonen@vtt.fi
authors.forename: Hetti
authors.initials: H
authors.lastname: Palonen
authors.forename: Folke
authors.initials: F
authors.lastname: Tjerneld
authors.forename: Guido
authors.initials: G
authors.lastname: Zacchi
authors.forename: Maija
authors.initials: M
authors.lastname: Tenkanen
mesh_terms: D000327:Adsorption; D001665:Binding Sites; D002480:Cellulase / Q000737:chemistry; D044602:Cellulases / Q000378:metabolism*; D004789:Enzyme Activation; D008031:Lignin / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013227:Steam; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology*; D014934:Wood
chemical_list: D013227:Steam; D008031:Lignin; D044602:Cellulases; C407024:endoglucanase Cel5A; D002480:Cellulase