title: Hydrolytic activity of free and immobilized cellulase.
abstract: Cellulase is an enzymatic complex which synergically promotes the degradation of cellulose to glucose. The adsorption behavior of cellulase from Trichoderma reesei onto Si wafers or amino-terminated surfaces was investigated by means of ellipsometry and atomic force microscopy (AFM) as a function of temperature. Upon increasing temperature from (24 +/- 1) to (60 +/- 1) degrees C, adsorption of cellulase became faster and more pronounced and the mean roughness of cellulase adsorbed layers increased. In the case of cellulase adsorbed onto Si wafers, Arrhenius's plot allowed us to estimate the adsorption energy as 24.2 kJ mol(-1). The hydrolytic activity of free cellulase and cellulase immobilized onto Si wafers was tested using cellulose dispersions as substrates. The incubation temperature ranged from (37 +/- 1) to (60 +/- 1) degrees C. The highest efficiency was observed at (60 +/- 1) degrees C. The amount of glucose produced by free cellulase was approximately 20% higher than that obtained from immobilized cellulase. However, immobilizing cellulase onto Si wafers proved to be advantageous because they could be reused six times while retaining their original activity level. Such an effect was attributed to surface hydration, which prevents enzyme denaturation. The hydrolytic activity of cellulase immobilized onto amino-terminated surfaces was slightly lower than that observed for cellulase adsorbed onto Si wafers, and reuse was not possible.
authors.affiliation: Instituto de Quimica, Universidade de Sao Paulo, Av. Prof. Lineu Prestes 748, 05508-900 Sao Paulo, Brazil.
authors.forename: Iris R M
authors.initials: IR
authors.lastname: Tébéka
authors.forename: Artur G L
authors.initials: AG
authors.lastname: Silva
authors.forename: Denise F S
authors.initials: DF
authors.lastname: Petri
mesh_terms: D000327:Adsorption; D055162:Biocatalysis; D002480:Cellulase / Q000378:metabolism* / Q000648:ultrastructure; D004800:Enzymes, Immobilized / Q000378:metabolism* / Q000648:ultrastructure; D005947:Glucose / Q000378:metabolism; D006868:Hydrolysis; D018625:Microscopy, Atomic Force; D014242:Trichoderma / Q000201:enzymology
chemical_list: D004800:Enzymes, Immobilized; D002480:Cellulase; D005947:Glucose