Fig. 3 | Nature Communications

Fig. 3

From: Single-molecule study of oxidative enzymatic deconstruction of cellulose

Fig. 3

Real-time observations of LPMO activity on cellulose nanocrystals and its effect on CBH I. a Partial detachment of cellulose fibrils from nanocrystals as a result of C1 oxidizing LPMO activity. LPMO molecule adsorbs (blue circle) to an intact crystal and after desorption a fibril loosened from the crystalline material (yellow frame) becomes visible. b By comparing forward (trace; green frames) and backward direction (retrace; blue frames) of AFM line scans, the loosened parts of the fibrils are shown to be moveable by the AFM tip. The fibril part associated with the cellulose nanocrystal, by contrast, was not moveable. Further examples of cellulose nanocrystal fibrillation by LMPO are shown in Supplementary Fig. 3. c LPMO enhances the adsorption of CBH I to cellulose nanocrystals, as shown in AFM images (compare (c) and Fig. 1b) and quantitative single-molecule analysis (d). d The ratio of enzyme molecules/top surface area analyzed for 16 cellulose nanocrystals is shown for the conditions that LPMO acted alone and CBH I was added afterwards. Experiment using CBH I alone is shown as reference. Real-time observations are also shown in Supplementary Movies 5 and 6. Besides showing enhanced enzyme adsorption, the movies reveal an increased portion of the adsorbed enzymes to be mobile on the cellulose surface when LPMO and CBH I were present together. Supplementary Movie 2, which shows CBH I acting alone, is used as reference. e Synergy between CBH I and LPMO during degradation of cellulose nanocrystals is shown. LPMO addition stimulated the CBH I adsorption and resulted in increased release of soluble sugars. Solid lines represent mean values and standard error is indicated by dashed lines. All scale bars are 10 nm

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