Ps AA9A, a C1-specific AA9 lytic polysaccharide monooxygenase from the white-rot basidiomycete Pycnoporus sanguineus

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BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS

PsAA9A, a C1-specific AA9 lytic polysaccharide monooxygenase from the white-rot basidiomycete Pycnoporus sanguineus Mercedes María Garrido 1,2 & Malena Landoni 3 & Federico Sabbadin 4 & María Pía Valacco 5 & Alicia Couto 3 & Neil Charles Bruce 4 & Sonia Alejandra Wirth 2 & Eleonora Campos 1 Received: 23 July 2020 / Revised: 4 September 2020 / Accepted: 14 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020

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 cellooligosaccharides 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.

Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10911-6) contains supplementary material, which is available to authorized users. * Eleonora Campos [email protected]

1

Instituto de Agrobiotecnología y Biología Molecular (IABIMO), Instituto Nacional de Tecnología Agropecuaria (INTA-CONICET), Los Reseros y Nicolas Repetto s/n (1686), Hurlingham, Buenos Aires, Argentina

2

Laboratorio de Agrobiotecnología, Instituto de Biodiversidad y Biología Experimental y Aplicada (IBBEA) CONICET-UBA, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (C1428EG), Buenos Aires, Argentina

3

Centro de Investigación en Hidratos de Carbono (CIHIDECAR), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (C1428EG), Buenos Aires, Argentina

Alicia Couto [email protected]

4

Neil Charles Bruce [email protected]

Centre for Novel Agricultural Products (CNAP), Department of Biology, University of York, York YO10 5DD, UK

5

Instituto de Química Biológica (IQUIBICEN), Facultad de Ciencias Exactas y Naturales, Universidad de Buen