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Communication Dans Un Congrès Année : 2014

New insights into mobility and affinity of engineered family 4 CBMs in bioinspired lignocellulosic assemblies

Résumé

Lignocellulose-acting enzymes are often appended to one or more carbohydrate binding modules (CBMs) which promote substrate targeting due to their variable substrate specificity and affinity. They thus play a central role in the enzymatic deconstruction of plant cell wall biomass, but their fine characterization is generally performed with pure isolated polymers which do not reflect the complex plant cell walls. In order to gain more information on the behavior and mobility of CBMs in complex 3D substrates, we have devised and used bioinspired model assemblies that contain some of the polymers and covalent interactions found in plant cell walls. These assemblies contain feruloylated arabinoxylans (FAXs) and cellulose nano-crystals (CNCs) at various concentrations in solution (free polymers) or in gels (cross-linked polymers). The three CBMs studied herein belong to family 4 and have been engineered so that they have different affinities and specificities toward FAX and CNC: CBM4-2 binds FAX and CNC, CBM X-2 binds only FAX and CBM G-4 binds neither FAX nor CNC. These CBMs have been produced as monomers and as dimers and then been grafted to a fluorophore so that their mobility in the bioinspired assemblies can be followed by measuring the fluorescence recovery after photobleaching. By varying three parameters related to the assemblies (CNC concentration, solution or gel state) and to the CBMs (oligomerisation state), we have been able to calculate an apparent affinity and to model the mobility of the CBMs. Overall, our results reveal that apparent affinity is drastically modified in a 3D polymer network in comparison to their behavior in solution, and parameters varied influence differently each CBM. This demonstrates that bioinspired assemblies may provide important information relevant for studies of carbohydrate active proteins such as the selection of appropriate CBM constructs to be further exploited in applications targeting complex carbohydrate substrates.
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Dates et versions

hal-01268552 , version 1 (04-02-2016)

Identifiants

  • HAL Id : hal-01268552 , version 1
  • PRODINRA : 262093

Citer

Gabriel Paës, Laura von Schantz, Mats Ohlin. New insights into mobility and affinity of engineered family 4 CBMs in bioinspired lignocellulosic assemblies. 247. ACS National Meeting - Chemistry & Materials for Energy, Mar 2014, Dallas, United States. ⟨hal-01268552⟩
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