Functional characterization of a lytic polysaccharide monooxygenase from the thermophilic fungus Myceliophthora thermophila.
KADOWAKI, Marco A.; VÁRNAI, Anikó; JAMESON, John-Kristian; LEITE, Ana E. T.; COSTA FILHO, Antonio José da; KUMAGAI, Patricia S.; PRADE, Rolf A.; POLIKARPOV, Igor; VINCENT, G. H. Eijsink.
KADOWAKI, Marco A.; VÁRNAI, Anikó; JAMESON, John-Kristian; LEITE, Ana E. T.; COSTA FILHO, Antonio José da; KUMAGAI, Patricia S.; PRADE, Rolf A.; POLIKARPOV, Igor; VINCENT, G. H. Eijsink.
 Abstract: Thermophilic fungi are a promising source of thermostable enzymes able to hydrolytically or oxidatively degrade plant cell wall components. Among these enzymes are lytic polysaccharide monooxygenases (LPMOs), enzymes capable of enhancing biomass hydrolysis through an oxidative mechanism. Myceliophthora thermophila (synonym Sporotrichum thermophile), an Ascomycete fungus, expresses and secretes over a dozen different LPMOs. In this study, we report the overexpression and biochemical study of a previously uncharacterized LPMO (MtLPMO9J) from M. thermophila M77 in Aspergillus nidulans. MtLPMO9J is a single-domain LPMO and has 63% sequence similarity with the catalytic domain of NcLPMO9C from Neurospora crassa. Biochemical characterization of MtLPMO9J revealed that it performs C4-oxidation and is active against cellulose, soluble cello-oligosaccharides and xyloglucan. Moreover, biophysical studies showed that MtLPMO9J is structurally stable at pH above 5 and at temperatures up to 50ÊC. Importantly, LC-MS analysis of the peptides after tryptic digestion of the recombinantly produced protein revealed not only the correct processing of the signal peptide and methylation of the N-terminal histidine, but also partial autoxidation of the catalytic center. This shows that redox conditions need to be controlled, not only during LPMO reactions but also during protein production, to protect LPMOs from oxidative damage. | |
| PLOS One |
| v. 13, n. 8, p. e0202148-1- e0202148-16 - Ano: 2018 |
| Fator de Impacto: 2,766 |
| http://dx.doi.org/10.1371/journal.pone.0202148 |  @article={002899302,author = {KADOWAKI, Marco A.; VÁRNAI, Anikó; JAMESON, John-Kristian; LEITE, Ana E. T.; COSTA FILHO, Antonio José da; KUMAGAI, Patricia S.; PRADE, Rolf A.; POLIKARPOV, Igor; VINCENT, G. H. Eijsink.},title={Functional characterization of a lytic polysaccharide monooxygenase from the thermophilic fungus Myceliophthora thermophila},journal={PLOS One},note={v. 13, n. 8, p. e0202148-1- e0202148-16},year={2018}} |