Heterologous Expression of <i>Phanerochaete chrysoporium</i> Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green

Son, Yu-Lim; Kim, Hyoun-Young; Thiyagarajan, Saravanakumar; Xu, Jing; Park, Seung-Moon

doi:10.5941/myco.2012.40.4.258

Cited by 8 publications

(7 citation statements)

References 18 publications

(29 reference statements)

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“…The optimum pH obtained under the tested conditions for both Pci GLOX enzymes was 6, which is in agreement with that previously reported for Ph GLOX ( 26 , 27 ). However, our results differ from those obtained by Son et al ( 25 ), who showed that the optimum pH for the recombinant Ph GLOX was 5. In our study, it was noticed that the enzymes were very sensitive to minor pH changes and they were less active at pH 5, which is a change of only 1 pH unit from the optimal pH.…”

Section: Discussioncontrasting

confidence: 99%

“…The two purified glycosylated proteins were similar in size to Ph GLOX (68 kDa), which was also found to be N-glycosylated, with glycosylation accounting for about 16% of the total protein mass ( 12 , 23 ). Contrary to the optimum temperature for Ph GLOX, which is 30°C ( 25 ), the optimum temperature for both Pci GLOX described here was 50°C, and activity was still detected at 75 to 80°C. The two Pci GLOX enzymes were found to be similarly affected by temperature and were relatively stable at 50°C after 4 h of incubation.…”

Section: Discussioncontrasting

confidence: 90%

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Heterologous Production and Characterization of Two Glyoxal Oxidases from Pycnoporus cinnabarinus

Daou

Piumi

Cullen

et al. 2016

Appl Environ Microbiol

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The genome of the white rot fungus Pycnoporus cinnabarinus includes a large number of genes encoding enzymes implicated in lignin degradation. Among these, three genes are predicted to encode glyoxal oxidase, an enzyme previously isolated from Phanerochaete chrysosporium. The glyoxal oxidase of P. chrysosporium is physiologically coupled to lignin-oxidizing peroxidases via generation of extracellular H2O2 and utilizes an array of aldehydes and α-hydroxycarbonyls as the substrates. Two of the predicted glyoxal oxidases of P. cinnabarinus, GLOX1 (PciGLOX1) and GLOX2 (PciGLOX2), were heterologously produced in Aspergillus niger strain D15#26 (pyrG negative) and purified using immobilized metal ion affinity chromatography, yielding 59 and 5 mg of protein for PciGLOX1 and PciGLOX2, respectively. Both proteins were approximately 60 kDa in size and N-glycosylated. The optimum temperature for the activity of these enzymes was 50°C, and the optimum pH was 6. The enzymes retained most of their activity after incubation at 50°C for 4 h. The highest relative activity and the highest catalytic efficiency of both enzymes occurred with glyoxylic acid as the substrate. The two P. cinnabarinus enzymes generally exhibited similar substrate preferences, but PciGLOX2 showed a broader substrate specificity and was significantly more active on 3-phenylpropionaldehyde. IMPORTANCE This study addresses the poorly understood role of how fungal peroxidases obtain an in situ supply of hydrogen peroxide to enable them to oxidize a variety of organic and inorganic compounds. This cooperative activity is intrinsic in the living organism to control the amount of toxic H2O2 in its environment, thus providing a feed-on-demand scenario, and can be used biotechnologically to supply a cheap source of peroxide for the peroxidase reaction. The secretion of multiple glyoxal oxidases by filamentous fungi as part of a lignocellulolytic mechanism suggests a controlled system, especially as these enzymes utilize fungal metabolites as the substrates. Two glyoxal oxidases have been isolated and characterized to date, and the differentiation of the substrate specificity of the two enzymes produced by Pycnoporus cinnabarinus illustrates the alternative mechanisms existing in a single fungus, together with the utilization of these enzymes to prepare platform chemicals for industry.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussioncontrasting

confidence: 90%

Heterologous Production and Characterization of Two Glyoxal Oxidases from Pycnoporus cinnabarinus

Daou

Piumi

Cullen

et al. 2016

Appl Environ Microbiol

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“…The maximum activity of MtGLOx was observed at pH 6.0 and 50 • C ( Figure 3A,B). These optimum conditions for MtGLOx were practically the same as described for the two glyoxal oxidases from P. cinnabarinus [27], but distinct from the P. chrysosporium enzyme (30 • C and pH 5.0) [38]. The residual activity after incubation at various temperatures was assayed at pH 6.0 using methylglyoxal as a substrate.…”

Section: Catalytic Properties Of Mtgloxmentioning

confidence: 92%

“…Moreover, some GLOxes are linked to the cell Wall Stress-responsive Component (WSC) domain that is predicted to be a cell-wall and membrane protein in yeast and fungi associated with cell wall integrity and stress responses [36,37]. Only three catalytic domains of GLOx enzymes from Phanaerochaete chrysosporium, Ustilago maydis, and Pycnoporus cinnabarinus fungi have been studied biochemically to date [26,27,34,38], but none were associated with such WSC domains. In the present study, we report the structural and functional characterization of a multidomain glyoxal oxidase from the thermo tolerant Myceliophthora thermophila M77 and show that 5-hydroxymethylfurfural as a new described substrate for this enzyme, demonstrating its potential in green enzymatic synthesis.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a New Glyoxal Oxidase from the Thermophilic Fungus Myceliophthora thermophila M77: Hydrogen Peroxide Production Retained in 5-Hydroxymethylfurfural Oxidation

et al. 2018

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Myceliophthora thermophyla is a thermophilic industrially relevant fungus that secretes an assortment of hydrolytic and oxidative enzymes for lignocellulose degradation. Among them is glyoxal oxidase (MtGLOx), an extracellular oxidoreductase that oxidizes several aldehydes and α-hydroxy carbonyl substrates coupled to the reduction of O2 to H2O2. This copper metalloprotein belongs to a class of enzymes called radical copper oxidases (CRO) and to the “auxiliary activities” subfamily AA5_1 that is based on the Carbohydrate-Active enZYmes (CAZy) database. Only a few members of this family have been characterized to date. Here, we report the recombinant production, characterization, and structure-function analysis of MtGLOx. Electron Paramagnetic Resonance (EPR) spectroscopy confirmed MtGLOx to be a radical-coupled copper complex and small angle X-ray scattering (SAXS) revealed an extended spatial arrangement of the catalytic and four N-terminal WSC domains. Furthermore, we demonstrate that methylglyoxal and 5-hydroxymethylfurfural (HMF), a fermentation inhibitor, are substrates for the enzyme.

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“…All these Primers were used to amplify cDNA and these were : 1. F: 5'-GCGAATTCATGTTGTCGCTGCTAGCCGTAGT-3 'R: 3'-CGTACGTACTCCAGGGTCGGCGGAGGGT-5' (Son et al, 2012).…”

Section: Pcr Primersmentioning

confidence: 99%

Detection the Effect of Cu++ on Transcription of glox Gene in Myceliophthora verrucosa Using RT-PCR

Khalil¹

2017

RJS

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Reverse transcriptase PCR was used to detect the transcription of the glox gene in a local isolate of M. verrucosa, in two different growth media. The results indicated the presence of gene transcription in this strain with no effect to Cu + on gene expression. This means that this fungus is suitable for use in various fields such as a bioremediation. This method depends on RNA to detect gene expression under various conditions. ‫ــ‬ ‫ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ‬

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Heterologous Expression of Phanerochaete chrysoporium Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green

Cited by 8 publications

References 18 publications

Heterologous Production and Characterization of Two Glyoxal Oxidases from Pycnoporus cinnabarinus

Heterologous Production and Characterization of Two Glyoxal Oxidases from Pycnoporus cinnabarinus

Characterization of a New Glyoxal Oxidase from the Thermophilic Fungus Myceliophthora thermophila M77: Hydrogen Peroxide Production Retained in 5-Hydroxymethylfurfural Oxidation

Detection the Effect of Cu++ on Transcription of glox Gene in Myceliophthora verrucosa Using RT-PCR

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