Dynamic modulation of proximity and activity of Enzymes immobilized on an Elastomeric surface – D-E-stancE

Enzymes play a crucial role in the bioeconomy, particularly when it comes to extracting value from lignocellulosic biomass. Some microorganisms have developed sophisticated strategies to efficiently degrade lignocellulose. These include the cellulosome, which is characterised by a panel of enzymes immobilised on a multi-protein complex linked to the outer cell membrane of the bacterium. In this configuration, the intrinsic flexibility of the complex enhances the synergy between the enzymes, which plays a key role in the efficiency of the cellulosome in degrading plant lignocellulosic biomass. The D-E-stancE project proposes a biomimetic approach to investigate the hypothesis that the distance between the enzymes and its dynamics during hydrolysis are responsible for the efficiency of the cellulosome. To this end, we propose an original approach involving the controlled immobilisation of enzymes on a custom-made elastic surface, which allows us to modulate both the enzymatic activity and the profile of products released, by simply stretching the surface mechanically. The enzymes used in our project are two pairs of cellulosomal enzymes from Ruminiclostridium cellulolyticum that will be co-immobilized in pairs, one active on arabinoxylan (xylanase GH10 and arabinofuranosidase GH43), and the other on cellulose (Cel48F and Cel9G). The project will improve our understanding of enzymatic synergy to improve the use of plant biomass and will also provide new insights into the field of enzyme immobilisation This interdisciplinary project brings together five partners with expertise in polymer chemistry for the production of elastic surfaces, enzymology for the production and characterisation of adapted chimeric enzymes, cellulosomal enzymes, advanced physico-chemical characterisation of grafted surfaces and AFM analysis to characterise the behaviour of enzymes under strain.