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Csarman, F; Gusenbauer, C; Wohlschlager, L; van Erven, G; Kabel, MA; Konnerth, J; Potthast, A; Ludwig, R.
(2021): Non-productive binding of cellobiohydrolase i investigated by surface plasmon resonance spectroscopy
CELLULOSE. 2021; 28(15): 9525-9545. FullText FullText_BOKU

Abstract:
Future biorefineries are facing the challenge to separate and depolymerize biopolymers into their building blocks for the production of biofuels and basic molecules as chemical stock. Fungi have evolved lignocellulolytic enzymes to perform this task specifically and efficiently, but a detailed understanding of their heterogeneous reactions is a prerequisite for the optimization of large-scale enzymatic biomass degradation. Here, we investigate the binding of cellulolytic enzymes onto biopolymers by surface plasmon resonance (SPR) spectroscopy for the fast and precise characterization of enzyme adsorption processes. Using different sensor architectures, SPR probes modified with regenerated cellulose as well as with lignin films were prepared by spin-coating techniques. The modified SPR probes were analyzed by atomic force microscopy and static contact angle measurements to determine physical and surface molecular properties. SPR spectroscopy was used to study the activity and affinity of Trichoderma reesei cellobiohydrolase I (CBHI) glycoforms on the modified SPR probes. N-glycan removal led to no significant change in activity or cellulose binding, while a slightly higher tendency for non-productive binding to SPR probes modified with different lignin fractions was observed. The results suggest that the main role of the N-glycosylation in CBHI is not to prevent non-productive binding to lignin, but probably to increase its stability against proteolytic degradation. The work also demonstrates the suitability of SPR-based techniques for the characterization of the binding of lignocellulolytic enzymes to biomass-derived polymers. [GRAPHICS] .
Authors BOKU Wien:
Csarman Florian
Gusenbauer Claudia
Konnerth Johannes
Ludwig Roland
Potthast Antje
Wohlschlager Lena
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Find related publications in this database (Keywords)
Cellobiohydrolase
Non-productive binding
Surface plasmon resonance
Biomass hydrolysis
Lignin
Glycosylation


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