Interaction and Kinetics of Oxidative Biomass Degrading Enzymes Resolved by High-Resolution Techniques
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.
lignocellulose degrading enzymes oxidoreductases extracellular fungal enzymes fluorescence mictroscopy scanning electrochemical microscopy
Publikationen
Project staff
Roland Ludwig
Assoc. Prof. Dipl.-Ing. Dr. Roland Ludwig
roland.ludwig@boku.ac.at
Tel: +43 1 47654-75216
Project Leader
01.03.2017 - 28.02.2022
Florian Csarman
Dipl.-Ing. Dr. Florian Csarman
florian.csarman@boku.ac.at
Tel: +43 1 47654-75213, 75214
Project Staff
01.03.2017 - 28.02.2022
Neus Gacias Amengual
Neus Gacias Amengual MSc.
neus.gacias@boku.ac.at
Project Staff
17.09.2018 - 28.02.2022
Kwankao Karnpakdee
Kwankao Karnpakdee MSc.
kwankao.karnpakdee@students.boku.ac.at
Tel: +43 1 47654-75252
Project Staff
01.05.2017 - 28.02.2022
Stefan Scheiblbrandner
Dipl.-Ing. Stefan Scheiblbrandner Ph.D.
Project Staff
01.03.2017 - 28.02.2022
Lena Wohlschlager
Dipl.-Ing. Dr. Lena Wohlschlager
lena.wohlschlager@boku.ac.at
Tel: +43 1 47654-89400
Project Staff
01.03.2017 - 30.09.2021