Agger, Jane W.6; Isaksen, Trine6; Várnai, Anikó6; Vidal Melgosa, Silvia7; Willats, William George Tycho7; Ludwig, Roland4; Horn, Svein J.6; Eijsink, Vincent G.H.6; Westereng, Bjørge8
1 Section for Plant Glycobiology, Department of Plant and Environmental Sciences, Faculty of Science, Københavns Universitet2 Forest, Nature and Biomass, Department of Geosciences and Natural Resource Management, Faculty of Science, Københavns Universitet3 Norwegian University of Life Sciences4 BOKU–University of Natural Resources and Life Sciences5 Molecular Plant Fysiology, Department of Plant Biology, Faculty of Life Sciences, Københavns Universitet6 Norwegian University of Life Sciences7 Section for Plant Glycobiology, Department of Plant and Environmental Sciences, Faculty of Science, Københavns Universitet8 Molecular Plant Fysiology, Department of Plant Biology, Faculty of Life Sciences, Københavns Universitet
The recently discovered lytic polysaccharide monooxygenases (LPMOs) are known to carry out oxidative cleavage of glycoside bonds in chitin and cellulose, thus boosting the activity ofwell-known hydrolytic depolymerizing enzymes. Because biomass-degrading microorganisms tend to produce a plethora of LPMOs, and considering the complexity and copolymeric nature of the plant cell wall, it has been speculated that some LPMOs may act on other substrates, in particular the hemicelluloses that tether to cellulose microfibrils. We demonstrate that an LPMO from Neurospora crassa, NcLPMO9C, indeed degrades various hemicelluloses, in particular xyloglucan. This activity was discovered using a glycan microarray-based screening method for detection of substrate specificities of carbohydrate-active enzymes, and further explored using defined oligomeric hemicelluloses, isolated polymeric hemicelluloses and cell walls. Products generated by NcLPMO9C were analyzed using high performance anion exchange chromatography and multidimensional mass spectrometry. We show that NcLPMO9C generates oxidized products from a variety of substrates and that its product profile differs from those of hydrolytic enzymes acting on the same substrates. The enzyme particularly acts on the glucose backbone of xyloglucan, accepting various substitutions (xylose, galactose) in almost all positions. Because the attachment of xyloglucan to cellulose hampers depolymerization of the latter, it is possible that the beneficial effect of the LPMOs that are present in current commercial cellulase mixtures in part is due to hitherto undetected LPMO activities on recalcitrant hemicellulose structures.
Proceedings of the National Academy of Sciences of the United States of America, 2014, Vol 111, Issue 17, p. 6287-6292