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Mode of action and active site of an extracellular peroxidase from Pleurotus ostreatus.

ABSTRACT: The properties of the haem environment of an extracellular peroxidase from Pleurotus ostreatus were studied by electronic absorption spectroscopy. A high-spin ferric form was predominant in the native enzyme and a high-spin ferrous form in the reduced enzyme. Cyanide was readily bound to the haem iron in the native form, thereby changing the enzyme to a low-spin cyano adduct. The electronic absorption spectra of the enzyme were similar to those of lignin peroxidase from Phanerochaete chrysosporium. Compound III of the enzyme was formed after the addition of an excess of H2O2 to the native enzyme, and thereafter spontaneously reverted to the native form. The enzyme oxidized 1-(3,5-dimethoxy-4-hydroxyphenyl)-2-(2-methoxyphenoxy)-1,3-dihydroxyp ropane in the presence of H2O2 to produce 1-(3,5-dimethoxy-4-hydroxyphenyl)-2-(2-methoxyphenoxy)-1-oxo-3-hydroxypr opane , 2,6-dimethoxyhydroquinone, 2-(2-methoxyphenoxy)-3-hydroxypropanal, 2-(2-methoxyphenoxy)-3-hydroxypropanoic acid, 2,6-dimethoxy-1,4-benzoquinone and guaiacol. A similar oxidation pattern was demonstrated with a one-electron oxidant, ammonium cerium(IV)nitrate. Free radicals were detected as intermediates of the enzyme-mediated oxidation of 1-(3,5-dimethoxy-5-hydroxyphenyl)-2-(2-methoxyphenoxy)-1,3-dihydroxyp ropane and acetosyringone. These results can be explained by the mechanisms involving an initial one-electron oxidation of the lignin substructure. This radical may undergo C alpha-C beta cleavage, C alpha-oxidation and alkyl-phenyl cleavage.

SUBMITTER: Han YH

PROVIDER: S-EPMC1217066 | biostudies-other | 1996 Mar

REPOSITORIES: biostudies-other

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Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus.

Project description:Manganese peroxidase (MnP) was produced from white rot edible mushroom Pleurotus ostreatus on the culture filtrate. The enzyme was purified to homogeneity using (NH4)2SO4 precipitation, DEAE-Sepharose and Sephadex G-100 column chromatography. The final enzyme activity achieved 81 U mL(-1), specific activity 78 U mg(-1) with purification fold of 130 and recovery 1.2% of the crude enzyme. SDS-PAGE indicated that the pure enzyme have a molecular mass of approximately 42 kDa. The optimum pH was between 4-5 and the optimum temperature was 25 °C. The pure MnP activity was enhanced by Mn(2+), Cu(2+), Ca(2+) and K(+) and inhibited by Hg(+2) and Cd(+2). H2O2 at 5 mM enhanced MnP activity while at 10 mM inhibited it significantly. The MnP-cDNA encoding gene was sequenced and determined (GenBank accession no. AB698450.1). The MnP-cDNA was found to consist of 497 bp in an Open Reading Frame (ORF) encoding 165 amino acids. MnP from P. ostreatus could detoxify aflatoxin B1 (AFB1) depending on enzyme concentration and incubation period. The highest detoxification power (90%) was observed after 48 h incubation at 1.5 U mL(-1) enzyme activities.

| S-EPMC4059287 | biostudies-literature

Pleurotus ostreatus manganese-dependent peroxidase silencing impairs decolourization of Orange II.

Project description:Decolourization of azo dyes by Pleurotus ostreatus, a white-rot fungus capable of lignin depolymerization and mineralization, is related to the ligninolytic activity of enzymes produced by this fungus. The capacity of P. ostreatus to decolourize the azo dye Orange II (OII) was dependent and positively co-linear to Mn(2+) concentration in the medium, and thus attributed to Mn(2+)-dependent peroxidase (MnP) activity. Based on the ongoing P. ostreatus genome deciphering project we identified at least nine genes encoding for MnP gene family members (mnp 1-9), of which only four (mnp 1-4) were previously known. Relative real-time PCR quantification analysis confirmed that all the nine genes are transcribed, and that Mn(2+) amendment results in a drastic increase in the transcript levels of the predominantly expressed MnP genes (mnp 3 and mnp 9), while decreasing versatile peroxidase gene transcription (mnp 4). A reverse genetics strategy based on silencing the P. ostreatus mnp 3 gene by RNAi was implemented. Knock-down of mnp 3 resulted in the reduction of fungal OII decolourization capacity, which was co-linear with marked silencing of the Mn(2+)-dependent peroxidase genes mnp 3 and mnp 9. This is the first direct genetic proof of an association between MnP gene expression levels and azo dye decolourization capacity in P. ostreatus, which may have significant implication on understanding the mechanisms governing lignin biodegradation. Moreover, this study has proven the applicability of RNAi as a tool for gene function studies in Pleurotus research.

| S-EPMC3815951 | biostudies-literature

Lignocellulose affects Mn2+ regulation of peroxidase transcript levels in solid-state cultures of Pleurotus ostreatus.

Project description:The effect of Mn2+ amendment on peroxidase gene expression was studied during Pleurotus ostreatus growth on cotton stalks. Four peroxidase-encoding genes were expressed differentially and in a manner different from that observed in defined media. Mn2+ affects mnp3 expression even 2 h after its addition to the cultures, suggesting a direct effect of the metal ion on expression.

| S-EPMC123952 | biostudies-literature

Direct oxidation of polymeric substrates by multifunctional manganese peroxidase isoenzyme from Pleurotus ostreatus without redox mediators.

Project description:VPs (versatile peroxidases) sharing the functions of LiP (lignin peroxidase) and MnP (manganese peroxidase) have been described in basidiomycetous fungi Pleurotus and Bjerkandera. Despite the importance of this enzyme in polymer degradation, its reactivity with polymeric substrates remains poorly understood. In the present study, we first report that, unlike LiP, VP from Pleurotus ostreatus directly oxidized two polymeric substrates, bovine pancreatic RNase and Poly R-478, through a long-range electron pathway without redox mediators. P. ostreatus produces several MnP isoenzymes, including the multifunctional enzyme MnP2 (VP) and a typical MnP isoenzyme MnP3. MnP2 (VP) depolymerized a polymeric azo dye, Poly R-478, to complete its catalytic cycle. Reduction of the oxidized intermediates of MnP2 (VP) to its resting state was also observed for RNase. RNase inhibited the oxidation of VA (veratryl alcohol) in a competitive manner. Blocking of the exposed tryptophan by N-bromosuccinimide inhibited the oxidation of RNase and VA by MnP2 (VP), but its Mn2+-oxidizing activity was retained, suggesting that Trp-170 exposed on an enzyme surface is a substrate-binding site both for VA and the polymeric substrates. The direct oxidation of RNase and Poly R by MnP2 (VP) is in sharp contrast with redox mediator-dependent oxidation of these polymers by LiP from Phanerochaete chrysosporium. Molecular modelling of MnP2 (VP) revealed that the differences in the dependence on redox mediators in polymer oxidation by MnP2 (VP) and LiP were explained by the anionic microenvironment surrounding the exposed tryptophan.

| S-EPMC1134804 | biostudies-other

Somatic transposition and meiotically driven elimination of an active helitron family in Pleurotus ostreatus.

Project description:Helitrons constitute a superfamily of DNA transposons that were discovered in silico and are widespread in most eukaryotic genomes. They are postulated to mobilize through a "rolling-circle" mechanism, but the experimental evidence of their transposition has been described only recently. Here, we present the inheritance patterns of HELPO1 and HELPO2 helitron families in meiotically derived progeny of the basidiomycete Pleurotus ostreatus. We found distorted segregation patterns of HELPO2 helitrons that led to a strong under-representation of these elements in the progeny. Further investigation of HELPO2 flanking sites showed that gene conversion may contribute to the elimination of such repetitive elements in meiosis, favouring the presence of HELPO2 vacant loci. In addition, the analysis of HELPO2 content in a reconstructed pedigree of subclones maintained under different culture conditions revealed an event of helitron somatic transposition. Additional analyses of genome and transcriptome data indicated that P. ostreatus carries active RNAi machinery that could be involved in the control of transposable element proliferation. Our results provide the first evidence of helitron mobilization in the fungal kingdom and highlight the interaction between genome defence mechanisms and invasive DNA.

| S-EPMC5397611 | biostudies-literature

Impact of transposable elements in Pleurotus ostreatus transcriptome profiles

Project description:Pleurotus ostreatus, also known as the oyster mushroom, is an active lignin decomposer in the forests. The genomes of the monokaryotic strains PC15 and PC9 have been used to characterize the content and distribution of transposable elements. This study analyzes the impact of transposable element insertions on the global transcriptome of P. ostreatus. The transcriptome of PC15 and PC9 has been analyzed in exponential growth during submerged fermentation in malt-yeast extract-sucrose medium RNAseq of two P. ostreatus strains: PC15 and PC9

2016-05-19 | E-GEOD-81586 | biostudies-arrayexpress

Degradation of Green Polyethylene by Pleurotus ostreatus.

Project description:We studied the biodegradation of green polyethylene (GP) by Pleurotus ostreatus. The GP was developed from renewable raw materials to help to reduce the emissions of greenhouse gases. However, little information regarding the biodegradation of GP discarded in the environment is available. P. ostreatus is a lignocellulolytic fungus that has been used in bioremediation processes for agroindustrial residues, pollutants, and recalcitrant compounds. Recently, we showed the potential of this fungus to degrade oxo-biodegradable polyethylene. GP plastic bags were exposed to sunlight for up to 120 days to induce the initial photodegradation of the polymers. After this period, no cracks, pits, or new functional groups in the structure of GP were observed. Fragments of these bags were used as the substrate for the growth of P. ostreatus. After 30 d of incubation, physical and chemical alterations in the structure of GP were observed. We conclude that the exposure of GP to sunlight and its subsequent incubation in the presence of P. ostreatus can decrease the half-life of GP and facilitate the mineralization of these polymers.

| S-EPMC4468114 | biostudies-literature

Potential of Pleurotus ostreatus mycelium for selenium absorption.

Project description:The aim of this study was to evaluate the effect of high selenium (Se) concentrations on morphophysiological and ultrastructural properties of Pleurotus ostreatus. Mycelium growth was good in media enriched with 5.0, 10.0, and 20.0 mg L(-1) of Se, concentration of 500.0 mg L(-1) strongly inhibited growth, and 1000.0 mg L(-1) was the minimum inhibitory concentration. Contrary to thin-walled, hyaline, branched, and anastomized hyphae with clamp-connections in the control, at Se concentrations of 100.0 and 500.0 mg L(-1), they were noticeably short, frequently septed and branched, with a more intensive extracellular matrix, and without clamp-connections. At high Se concentrations, hyphae with intact membrane, without cellular contents, with a high level of vacuolization, and with numerous proteinaceous bodies were observed. Biomass yield ranged between 11.8 g L(-1), in the control, and 6.8 g L(-1), at an Se concentration of 100.0 mg L(-1), while no production was detected at a concentration of 500.0 mg L(-1). Se content in the mycelia reached a peak (938.9 μg g(-1)) after cultivation in the medium enriched with Se at the concentration of 20.0 mg L(-1), while the highest absorption level (53.25%) was found in the medium enriched with 5.0 mg L(-1) Se.

| S-EPMC4066853 | biostudies-other

Impact of transposable elements in Pleurotus ostreatus transcriptome profiles

2016-05-19 | GSE81586 | GEO

Transposon-associated epigenetic silencing during Pleurotus ostreatus life cycle

Project description:In this study, we analyzed the genome-wide epigenetic and transcriptional patterns of the white-rot basidiomycete Pleurotus ostreatus throughout its life cycle. Our results performed by using high-throughput sequencing analyses revealed that strain-specific DNA methylation profiles are primarily involved in the repression of transposon activity, and suggest that 21 nt small RNAs play a key role in transposon silencing.Furthermore, we provide evidence that transposon-associated DNA methylation, but not sRNA production, is directly involved in the silencing of genes surrounded by transposons. Finally, we identified key genes activated in the fruiting process through the comparative analysis of transcriptomes.

2018-05-15 | GSE112588 | GEO

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