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Cell‐surface exposure of a hybrid 3‐cohesin scaffoldin allowing the functionalization of Escherichia coli envelope

Abstract : Cellulosomes are large plant cell wall degrading complexes secreted by some anaerobic bacteria. They are typically composed of a major scaffolding protein containing multiple receptors called cohesins, which tightly anchor a small complementary module termed dockerin harbored by the cellulosomal enzymes. In the present study, we have successfully cell surface exposed in Escherichia coli a hybrid scaffoldin, Scaf6, fused to the curli protein CsgA, the latter is known to polymerize at the surface of E. coli to form extracellular fibers under stressful environmental conditions. The C‐terminal part of the chimera encompasses the hybrid scaffoldin composed of three cohesins from different bacterial origins and a carbohydrate‐binding module targeting insoluble cellulose. Using three cellulases hosting the complementary dockerin modules and labeled with different fluorophores, we have shown that the hybrid scaffoldin merged to CsgA is massively exposed at the cell surface of E. coli and that each cohesin module is fully operational. Altogether these data open a new route for a series of biotechnological applications exploiting the cell‐surface exposure of CsgA‐Scaf6 in various industrial sectors such as vaccines, biocatalysts or bioremediation, simply by grafting the small dockerin module to the desired proteins before incubation with the engineered E. coli.
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Submitted on : Friday, November 13, 2020 - 12:13:30 PM
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Vita et al 2020 Biotechnol Bio...
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Nicolas Vita, Romain Borne, Henri‐pierre Fierobe. Cell‐surface exposure of a hybrid 3‐cohesin scaffoldin allowing the functionalization of Escherichia coli envelope. Biotechnology and Bioengineering, Wiley, 2020, 117 (3), pp.626-636. ⟨10.1002/bit.27242⟩. ⟨hal-02487345⟩



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