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. 2016 Jan 14;6:19392. doi: 10.1038/srep19392

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(a) Binding of CBM_C5614-1 variants to soluble polysaccharides. Proteins and BSA were separated using non-denaturing polyacrylamide gels containing 0.1% (wt/vol) soluble polysaccharides. A gel without polysaccharides served as a control (CK). M: BSA non-binding control; 1: CBM_C5614-1; 2: F372A; 3: W380A; 4: Y404A; 5: F408A; 6: Y411A; 7: F416A; 8: Y418A; 9: W423A; 10: F462A; 11: Y467A. (b) Binding of CBM_C5614-1 variants to insoluble polysaccharides. 30 μg of purified protein was incubated with 200 μl 4% (wt/vol) insoluble polysaccharide including Avicel (A), ASC (B), insoluble birch wood xylan (C), mannan (D), lichenan (E) and raw starch from cassava (F). The same amount of protein used in the binding assay but without polysaccharide was included as a control (CK). After centrifugation, unbound protein in the supernatant (lane 1) and bound proteins in the precipitate (lane 2) were analyzed by SDS-PAGE.