The interactions between films of cellulose and cellulase enzymes were monitored using a quartz crystal microbalance (QCM). Real-time measurements of the coupled contributions of enzyme binding and hydrolytic reactions were fitted to a kinetic model that described closely significant cellulase activities. The proposed model combines simple Boltzmann sigmoidal and 1 - exp expressions. The obtained kinetics parameters were proven to be useful to discriminate the effects of incubation variables and also to perform enzyme screening. Furthermore, it is proposed that the energy dissipation of a film subject to enzymatic hydrolysis brings to light its structural changes. Overall, it is demonstrated that the variations registered in QCM frequency and dissipation of the film are indicative of mass and morphological transformations due to enzyme activities; these include binding phenomena, progressive degradation of the cellulose film, existence of residual, recalcitrant cellulose fragments, and the occurrence of other less apparent changes throughout the course of incubation.

Model films of native cellulose nanofibrils, which contain both crystalline cellulose I and amorphous domains, were used to investigate the dynamics and activities of cellulase enzymes. The enzyme binding and degradation of nanofibril films were compared with those for other films of cellulose, namely, Langmuir-Schaefer and spin-coated regenerated cellulose, as well as cellulose nanocrystal cast films. Quartz crystal microbalance with dissipation (QCM-D) was used to monitor the changes in frequency and energy dissipation during incubation at varying enzyme concentrations and experimental temperatures. Structural and morphological changes of the cellulose films upon incubation with enzymes were evaluated by using atomic force microscopy. The QCM-D results revealed that the rate of enzymatic degradation of the nanofibril films was much faster compared to the other types of cellulosic films. Higher enzyme loads did not dramatically increase the already fast degradation rate. Real-time measurements of the coupled contributions of enzyme binding and hydrolytic reactions were fitted to an empirical model that closely described the cellulase activities. The hydrolytic potential of the cellulase mixture was found to be considerably affected by the nature of the substrates, especially their crystallinity and morphology. The implications of these observations are discussed in this report.

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