Carbohydrate-binding property of a cell wall integrity and stress response component (WSC) domain of an alcohol oxidase from the rice blast pathogen Pyricularia oryzae

Oide, Shinichi; Tanaka, Yoshihiko; Watanabe, Akira; Inui, Masayuki

doi:10.1016/j.enzmictec.2019.02.009

Cited by 38 publications

(48 citation statements)

References 38 publications

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“…Taken together, these results indicate that AlcOx activation is not dependent on highly efficient peroxidase activity. We also heterologously produced the Perox-AlcOx pair from the rice blast pathogen M. oryzae, of which MorAlcOx was recently confirmed to be a primary alcohol oxidase 33 .…”

Section: Tandem Perox-alcox Oxidize Plant Long Chain Alcoholsmentioning

confidence: 99%

Unravelling the role of alcohol copper radical oxidases in fungal plant pathogens

Bissaro

Kodama

Hage

et al. 2021

Preprint

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Copper radical oxidases (CRO) form a class of enzymes with a longstanding history encompassing diverse substrate specificities. While the biological function of most CROs remains unknown, we observed that CROs active on aliphatic alcohols are found only in fungal plant pathogens. Here, we unveil the role of these CROs and the identity of their natural redox partner, a haem-iron peroxidase. Combining multiscale approaches, we report that Colletotrichum and Magnaporthe appressoria (specialized cells that puncture the plant cuticles) co-secrete this pair of metalloenzymes early during penetration. We show in vivo that mutant appressoria lacking either or both enzymes have impaired penetration ability and pathogenicity. We reveal in vitro a finely-tuned enzyme interplay is responsible for the oxidation of plant cuticular long-chain alcohols into aldehyde products, suggested to act as key molecular signals in the fungal infection machinery. Our results open new avenues to design oxidase-specific inhibitors as anti-penetrants for crop protection.

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Section: Tandem Perox-alcox Oxidize Plant Long Chain Alcoholsmentioning

confidence: 99%

Unravelling the role of alcohol copper radical oxidases in fungal plant pathogens

Bissaro

Kodama

Hage

et al. 2021

Preprint

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“…Each node is colored according to its modularity. Catalytic modules are shown in white, carbohydrate binding modules are in green5 , PAN_1 domains are blue6 , WSC are brown7 and GPI anchor are yellow (B). CgrAlcOx = Colletotrichum graminicola alcohol oxidase, CglAlcOx = Colletotrichum gloeosporioides alcohol oxidase, CgrRafOx = Colletotrichum graminicola raffinose oxidase, PruAA5_2A = Penicillium rubens Wisconsin 54-1255 AA5_2 oxidase, FgrGalOx = Fusarium graminearum galactose oxidase, ChiAlcOx = Colletotrichum higginsianum alcohol oxidase and PorAlcOx = Pyricularia oryzae alcohol oxidase.…”

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confidence: 99%

Discovery of a Fungal Copper Radical Oxidase with High Catalytic Efficiency toward 5-Hydroxymethylfurfural and Benzyl Alcohols for Bioprocessing

et al. 2020

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Alternatives to petroleum-based chemicals are highly sought-after for ongoing efforts to reduce the damaging effects of human activity on the environment. Copper radical oxidases from Auxiliary Activity Family 5/Subfamily 2 (AA5_2) are attractive biocatalysts because they oxidize primary alcohols in a chemoselective manner without complex organic cofactors. However, despite numerous studies on canonical galactose oxidases (GalOx, EC 1.1.3.9) and engineered variants, and the recent discovery of a Colletotrichum graminicola copper radical alcohol oxidase (AlcOx, EC 1.1.3.13), the catalytic potentials of very few AA5_2 members have been characterized. Guided by the sequence similarity network and phylogenetic analyses, we targeted a distinct paralog from the fungus C. graminicola as a representative member of a large uncharacterized subgroup of AA5_2. Through recombinant production and detailed kinetic analysis, we demonstrated that this enzyme is weakly active toward carbohydrates but efficiently catalyzes the oxidation of aryl alcohols to the corresponding aldehydes. As such, this represents the initial characterization of a demonstrable aryl alcohol oxidase (AAO, EC 1.1.3.7) in AA5, an activity which is classically associated with flavin-dependent glucose-methanol-choline (GMC) oxidoreductases of Auxiliary Activity Family 3 (AA3). X-ray crystallography revealed a distinct multidomain architecture comprising an N-terminal PAN domain abutting a canonical AA5 seven-bladed propeller catalytic domain. Of direct relevance to biomass processing, the wild-type enzyme exhibits the highest activity on the primary alcohol of 5-hydroxymethylfurfural (HMF), a product of significant interest in the lignocellulosic biorefinery concept. Thus, the chemoselective oxidation of HMF to 2,5-diformylfuran (DFF) by C. graminicola aryl alcohol oxidase (CgrAAO) from AA5 provides a fundamental building block for chemistry via biotechnology.

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“…In contrast, HyBPOX2s from the same genomes contain besides the N-terminal peroxidase domain (of the same length and also classified in Pfam00141) a single middle domain with a closely related CBM34 sugar-binding motif. Further, they possess also two or even three C-terminal water soluble carbohydrate binding (WSC) domains ( Figure 2 c) known as cell-wall stress sensors [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

Hybrid Heme Peroxidases from Rice Blast Fungus Magnaporthe oryzae Involved in Defence against Oxidative Stress

et al. 2020

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Hybrid B heme peroxidases are recently discovered unique oxidoreductases present solely in the fungal kingdom. We have investigated two typical representatives from Magnaporthe oryzae—one of the most dangerous phytopathogens known as a causal agent of the rice blast disease. First, we focused on native expression of two detected hyBpox paralogs by the means of reverse-transcription quantitative real-time PCR. Our results indicate a 7-fold induction of the MohyBpox1 transcript in a medium with H2O2 and a 3-fold induction in a medium with peroxyacetic acid. For the MohyBpox2 paralog the induction patterns were up to 12-fold and 6.7-fold, respectively. We have successfully expressed the shorter gene, MohyBpox1, heterologously in Pichia pastoris for detailed characterization. Observed biochemical and biophysical properties of the highly purified protein reveal that a typical HyBPOX is significantly different from previously investigated APx-CcP hybrids. This newly discovered secretory peroxidase reveals a Soret maximum at 407 nm, Q bands at 532 and 568 nm, CT band at 625 nm and a purity number of 1.48. Electron paramagnetic resonance (EPR) analysis suggests a mixture of high and low spin species in the ferric state dependent on calcium contents. Steady-state kinetic data reveal the highest peroxidase activity with ABTS, 5-aminosalycilate and efficient oxidation of tyrosine. MoHyBPOX1 as a fusion protein consists of two domains. The longer conserved N-terminal peroxidase domain is connected with a shorter C-terminal domain containing a carbohydrate binding motif of type CBM21. We demonstrate the capacity of MoHyBPOX1 to bind soluble starch efficiently. Potential involvement of hybrid peroxidases in the pathogenicity of M. oryzae is discussed.

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Carbohydrate-binding property of a cell wall integrity and stress response component (WSC) domain of an alcohol oxidase from the rice blast pathogen Pyricularia oryzae

Cited by 38 publications

References 38 publications

Unravelling the role of alcohol copper radical oxidases in fungal plant pathogens

Unravelling the role of alcohol copper radical oxidases in fungal plant pathogens

Discovery of a Fungal Copper Radical Oxidase with High Catalytic Efficiency toward 5-Hydroxymethylfurfural and Benzyl Alcohols for Bioprocessing

Hybrid Heme Peroxidases from Rice Blast Fungus Magnaporthe oryzae Involved in Defence against Oxidative Stress

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