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New enZymes for BIOMass carbohydrate deconstruction

Periodic Report Summary 1 - ZYBIOM (New enZymes for BIOMass carbohydrate deconstruction)

Sustainable production of fuels and chemicals from biomass (e.g. lignocellulose) is essential considering the societal and political demands for alternatives to petrochemical commodities. One of the major bottlenecks to achieve economically viable processes is efficient enzymatic deconstruction of plant biomass, since the structure and composition of lignocellulose make degradation difficult The most promising strategy to circumvent this recalcitrance is the identification of new enzymes with activities complementary to existing enzyme cocktails. In this context, the ZyBiom project aims at (i) implementing a new approach to screen for lignocellulose-targetting activities by combining classical assays and fibre surface analysis (ToF-SIMS) and (ii) identifying new enzymes of interest from fungal diversity and exploration of metagenomes originating from mammal digestive systems.

In the first period, the following results have been achieved :
• The methodology combining ToF-SIMS analysis and high throughput DNS assay has been implemented at UofT, training of Marie Couturier on ToF-SIMS handling has been performed ;
• Improvement of ToF-SIMS application to biomass characterization has started with experiments aiming at discriminating different polysaccharide peak signatures and at identifying signals originating not only from polysaccharide deconstruction but also modifications;
• Screening of metagenome-originating enzymes has been performed and led to the identification of new beta-glucosidases ;
• Study of microcosms originating from canadian mammal digestive systems has started and led to the identification of families of enzymes (CAZymes) enriched in specific conditions
• Recovery of microcosm secreted enzymes has been established and exploration of their activities has started
• Exploration of fungal secretomes has been performed through transcriptomic and proteomic analyses as well as functional characterization of enzyme cocktails. This work led to better understanding of fungal strategies to efficiently degrade softwood and to the identification of potentially key enzymes for lignin and polysaccharide conversion.

In the meantime, transfer and dissemination of knowledge have been achieved by :
• Training of students at UofT on handling and programming the Liquid Handler ;
• Mentoring students (one PhD student and one master student) currently involved at UofT in the characterization of microcosm activities on biomass ;
• Publication of results in journals with high impact factors, as well as preparation of publication for the newest results ;
• Presentations of results at international conferences and national or regional workshops via oral or poster presentations.

The expected results obtained from both fungal and microcosm approaches of ZyBiom project will allow not only the identification of specific enzymes of interest for lignin and polysaccharide deconstruction, but will also bring new fundamental information to the scientific community about microbial strategies for biomass deconstruction.