Abstract
Woody biomass represents an important source of carbon on earth, and its global recycling is highly dependent on Agaricomycetes fungi. White-rot Basidiomycetes are a very important group in this regard, as they possess a large and diverse enzymatic repertoire for biomass decomposition. Among these enzymes, the recently discovered lytic polysaccharide monooxygenases (LPMOs) have revolutionized biomass processing with their novel oxidative mechanism of action. The strikingly high representation of LPMOs in fungal genomes raises the question of their functional versatility. In this work, we studied an AA9 LPMO from the white-rot basidiomycete Pycnoporus sanguineus, PsAA9A. Successfully produced as a recombinant secreted protein in Pichia pastoris, PsAA9A was found to be a C1-specific LPMO active on cellulosic substrates, generating native and oxidized cello-oligosaccharides in the presence of an external electron donor. PsAA9A boosted cellulolytic activity of glysoside hydrolases from families GH1, GH5, and GH6.This study serves as a starting point towards understanding the functional versatility and biotechnological potential of this enzymatic family, highly represented in wood decay fungi, in Pycnoporus genus.
Key points
• PsAA9A is the first AA9 from P. sanguineus to be characterized.
• PsAA9A has activity on cellulose, producing C1-oxidized cello-oligosaccharides.
• Boosting activity with GH1, GH5, and GH6 was proven.
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Data availability
Data of P. sanguineus PsAA9A synthetic coding sequence was deposited at DDBJ/EMBL/GenBank under the accession number MT076044.
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Acknowledgments
Authors thank Dr. Leonardo Gomez and Rachel Hallam for invaluable technical assistance.
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This work was supported by the Grants PICT2016-4695 (from the National Agency for Science and Technology Promotion from Argentina, ANPCyT) and Grant 20020130100476BA (from the University of Buenos Aires). The Ultraflex II (Bruker, Billerica, USA) TOF/TOF mass spectrometer was supported by ANPCyT Grant PME 125 (CEQUIBIEM). ML, AC, MPV, SAW and EC are Research Career Scientists of the National Research Council of Argentina (CONICET). NCB and EC acknowledge funding from the N8 AgriFood catalyst programme.
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EC, NCB, and SW conceived and designed research. MG, ML, and FS conducted experiments. PV and AC contributed analytical tools and analyzed data. MG wrote the manuscript. All authors read and approved the manuscript.
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Garrido, M.M., Landoni, M., Sabbadin, F. et al. PsAA9A, a C1-specific AA9 lytic polysaccharide monooxygenase from the white-rot basidiomycete Pycnoporus sanguineus. Appl Microbiol Biotechnol 104, 9631–9643 (2020). https://doi.org/10.1007/s00253-020-10911-6
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DOI: https://doi.org/10.1007/s00253-020-10911-6