Heterologous expression and characterization of a MoAA16 polysaccharide monooxygenase from the rice blast fungus Magnaporthe oryzae

Abstract

Background: Cellulose is an organic carbon source that can be used as a sustainable alternative for energy, materials, and chemicals. However, the substantial challenge of converting it into soluble sugars remains a major obstacle in its use as a biofuel and chemical feedstock. A new class of enzymes knowns as copper-dependent polysaccharide monooxygenases (PMOs) or lytic polysaccharide monooxygenases (LPMOs) can break down polysaccharides such as cellulose, chitin, and starch through oxidation. This process enhances the efficiency of cellulose degradation by cellulase. Results: The genome of the fungus Magnaporthe oryzae, the causal agent of rice blast disease, contains the MGG_00245 gene, which encodes a putative PMO referred to as MoAA16. MoAA16 has been found to be highly expressed in planta during the early stages of fungal infection. The gene was optimized for heterologous expression in Pichia pastoris, and its oxidative cleavage activity on cellulose was characterized by analyzing soluble oligosaccharide products using highperformance anion exchange chromatography (HPAEC-PAD). The reaction catalyzed by MoAA16 requires 2 electrons from an electron donor, such as ascorbic acid, and aerobic conditions. It primarily produces Glc1 to Glc4 oligosaccharides, as well as oxidized cellobionic and cellotrionic acids. MoAA16 has been observed to enhance cellulase hydrolysis on phosphoric acid swollen cellulose (PASC) substrate, resulting in the production of more monosaccharide products. Conclusions: Our findings reveal the successful heterologous expression of MoAA16 in P. pastoris and its cellulose-active PMO properties. These results highlight the potential of MoAA16 as a promising candidate for applications in biofuel production and chemical synthesis. How to cite: Nguyen HM, Le LQ, Sella L, et al. Heterologous expression and characterization of a MoAA16 polysaccharide monooxygenase from the rice blast fungus Magnaporthe oryzae. Electron J Biotechnol 2023. https://doi.org/10.1016/j.ejbt.2023.06.002.

Publication DOI: https://doi.org/10.1016/j.ejbt.2023.06.002
Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
College of Health & Life Sciences
Aston University (General)
Funding Information: This work is part of the Scientific and Technological Cooperation Agreement between the Ministry of Science and Technology of Vietnam and the Italian Ministry of Foreign Affairs and International Cooperation (grant No. NĐT.36.ITA/18). We also acknowledge
Additional Information: Funding Information: This work is part of the Scientific and Technological Cooperation Agreement between the Ministry of Science and Technology of Vietnam and the Italian Ministry of Foreign Affairs and International Cooperation (grant No. NĐT.36.ITA/18). We also acknowledge the UK Biotechnology and Biological Sciences Research Council through the Global Challenges Research Fund Project, CAPRI-BIO (BB/P022685/1). Publisher Copyright: 2023 The Authors. Pontificia Universidad Católica de Valparaíso. Production and hosting by Elsevier B.V This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Uncontrolled Keywords: Cellulose,Enzymatic activity,Fungal infection,Heterologous expression,Magnaporthe oryzae,Monosaccharide,Oxidative cleavage,Pichia pastoris,Polysaccharide monooxygenase,Protein expression,Rice blast fungus,Biotechnology,Applied Microbiology and Biotechnology
Publication ISSN: 0717-3458
Last Modified: 02 Dec 2024 08:59
Date Deposited: 11 Sep 2023 13:17
Full Text Link:
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
https://www.sci ... 0210?via%3Dihub (Publisher URL)
PURE Output Type: Article
Published Date: 2023-11
Published Online Date: 2023-08-28
Accepted Date: 2023-06-28
Authors: Nguyen, Hung M.
Le, Loan Q.
Sella, Luca
Broadbent, Luke M.
Bill, Roslyn M. (ORCID Profile 0000-0003-1331-0852)
Vu, Van V.

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