Xylan-cellulose thin film platform for assessing xylanase activity
2022
Schaubeder, Jana B. | Ravn, Jonas L. | Orzan, Eliott J.Q. | Manfrão-Netto, João H.C. | Geijer, Cecilia | Nypelö, Tiina | Spirk, Stefan
Enzymatic degradation of plant polysaccharide networks is a complex process that involves disrupting an intimate assembly of cellulose and hemicelluloses in fibrous matrices. To mimic this assembly and to elucidate the efficiency of enzymatic degradation in a rapid way, models with physicochemical equivalence to natural systems are needed. Here, we employ xylan-coated cellulose thin films to monitor the hydrolyzing activity of an endo-1,4-β-xylanase. In situ surface plasmon resonance spectroscopy (SPRS) revealed a decrease in xylan areal mass ranging from 0.01 ± 0.02 to 0.52 ± 0.04 mg·m⁻². The extent of digestion correlates to increasing xylanase concentration. In addition, ex situ determination of released monosaccharides revealed that incubation time was also a significant factor in degradation (P > 0.01). For both experiments, atomic force microscopy confirmed the removal of xylans from the cellulose thin films. We provide a new model platform that offers nanoscale sensitivity for investigating biopolymer interactions and their susceptibility to enzymatic hydrolysis.
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