Investigating lytic polysaccharide monooxygenase-assisted wood cell wall degradation with microsensors
Hucheng Chang, Neus Gacias Amengual, Alexander Botz, Lorenz Schwaiger, Daniel Kracher, Stefan Scheiblbrandner, Florian Csarman, Roland Ludwig
Abstract
Abstract Lytic polysaccharide monooxygenase (LPMO) supports biomass hydrolysis by increasing saccharification efficiency and rate. Recent studies demonstrate that H 2 O 2 rather than O 2 is the cosubstrate of the LPMO-catalyzed depolymerization of polysaccharides. Some studies have questioned the physiological relevance of the H 2 O 2 -based mechanism for plant cell wall degradation. This study reports the localized and time-resolved determination of LPMO activity on poplar wood cell walls by measuring the H 2 O 2 concentration in their vicinity with a piezo-controlled H 2 O 2 microsensor. The investigated Neurospora crassa LPMO binds to the inner cell wall layer and consumes enzymatically generated H 2 O 2 . The results point towards a high catalytic efficiency of LPMO at a low H 2 O 2 concentration that auxiliary oxidoreductases in fungal secretomes can easily generate. Measurements with a glucose microbiosensor additionally demonstrate that LPMO promotes cellobiohydrolase activity on wood cell walls and plays a synergistic role in the fungal extracellular catabolism and in industrial biomass degradation.