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Interaction and Kinetics of Oxidative Biomass Degrading Enzymes Resolved by High-Resolution Techniques

Project Leader
Ludwig Roland, Project Leader
Duration:
01.03.2017-28.02.2022
Programme:
Horizon 2020 - Excellent Science - ERC Consolidator Grant (CoG)
Type of Research
Basic Research

Further information: https://erc.europa.eu/

Staff
Kittl Roman, Project Staff (bis 31.07.2019)
Kracher Daniel, Project Staff
Scheiblbrandner Stefan, Project Staff
Csarman Florian, Project Staff
Ma Su, Project Staff
Wohlschlager Lena, Project Staff
Obermann Tobias, Project Staff (bis 30.06.2018)
Chang Hucheng, Project Staff
Gacias Amengual Neus, Project Staff
Botz Alexander, Project Staff
BOKU Research Units
Institute of Food Technology (LMT)
Funded by
Commission of the European Communities, Rue de la Loi, Brussels, European Union
Abstract
The mission of OXIDISE is to resolve authentic enzymatic activities of lignocellulose degrading oxidoreductases when bound onto their polymeric substrates and to elucidate their interaction. To this purpose, high-resolution techniques will be developed in the project. Fungal oxidoreductases involved in lignocellulose processing attracted great attention in the last years - like the discovery of oxidative cellulose degradation by lytic polysaccharide monooxygenase (LPMO). Over 90% of biomass degrading fungal genomes contain oxidoreductases (LPMO, cellobiose dehydrogenase, laccase, lignin peroxidase, aryl alcohol oxidase,…) involved in the oxidative cleavage of the recalcitrant biopolymers cellulose, hemicellulose or lignin. The elucidation of these enzyme mechanisms, interactions and kinetics is the key to understand fungal physiology and optimise biomass saccharification and biorefineries.
To circumvent typical problems associated with heterogeneous reactions, assaying techniques should have a high spatial and temporal resolution. OXIDISE will develop and apply techniques based on microelectrodes, scanning electron microscopy (SECM), surface-enhanced raman scattering (SERS) and microscopic fluorescence techniques to pursue five objectives: to 1) develop enzyme-modified electrodes for the detection of lignocellulose oxidising enzymes or their products. 2) miniaturise and assemble microelectrode arrays with a high spatial resolution. 3) transfer microelectrode modifications to SECM for increased spatial resolution (~10 nm) and scanning of areas. 4) investigate the interaction of oxidoreductases on polymeric substrates, e.g. the interaction of CDH/LPMO or more complex enzyme systems. 5) transfer the developed techniques to wood samples and growing fungal hyphae and their secretome. OXIDISE takes a new approach to provide crucial insight into the function of important enzymes, which so far has been somewhat neglected, possibly because of the involved experimental challenges.
Keywords
Electrochemistry; Catalysis; Molecular biology; Mycology; Ultrastructure research; Renewable resources; Food biotechnology;
scanning electrochemical microscopy; extracellular fungal enzymes; fluorescence mictroscopy; lignocellulose degrading enzymes; oxidoreductases;
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