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Monoclonal Antibody Services> | The binding of each cellulosomal component to scaffoldin was analyzed by an electrophoretic mobility shift assay of miniscaffoldin (Δ CipA) using native PAGE on 4–20% gradient gels with western blot analysis using an anti-Strep tag monoclonal antibody (THE NWSHPQFEK Tag antibody ; GenScript, USA) as described previously9. | Get A Quote |
The cellulosome is a supramolecular multienzyme complex comprised of a wide variety of polysaccharide-degrading enzymes and scaffold proteins. The cellulosomal enzymes that bind to the scaffold proteins synergistically degrade crystalline cellulose. Here, we report in vitro reconstitution of the Clostridium thermocellum cellulosome from 40 cellulosomal components and the full-length scaffoldin protein that binds to nine enzyme molecules. These components were each synthesized using a wheat germ cell-free protein synthesis system and purified. Cellulosome complexes were reconstituted from 3, 12, 30, and 40 components based on their contents in the native cellulosome. The activity of the enzyme-saturated ... More