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Grzegorz Janusz - One of the best experts on this subject based on the ideXlab platform.

  • Lighting Conditions Influence the Dynamics of Protease Synthesis and Proteasomal Activity in the White Rot Fungus Cerrena Unicolor.
    Biomolecules, 2020
    Co-Authors: Anna Pawlik, Marta Ruminowicz-stefaniuk, Andrzej Mazur, Magdalena Jaszek, Anita Swatek, Beata Ciołek, Grzegorz Janusz
    Abstract:

    Recent transcriptomic and biochemical studies have revealed that light influences the global gene expression profile and metabolism of the white-rot fungus Cerrena Unicolor. Here, we aimed to reveal the involvement of proteases and ubiquitin-mediated proteolysis by the 26S proteasome in the response of this fungus to white, red, blue and green lighting conditions and darkness. The changes in the expression profile of C. Unicolor genes putatively engaged in proteolysis were found to be unique and specific to the applied wavelength of light. It was also demonstrated that the activity of proteases in the culture fluid and mycelium measured using natural and synthetic substrates was regulated by light and was substrate-dependent. A clear influence of light on protein turnover and the qualitative and quantitative changes in the hydrolytic degradation of proteins catalyzed by various types of proteases was shown. The analysis of activity associated with the 26S proteasome showed a key role of ATP-dependent proteolysis in the initial stages of adaptation of fungal cells to the stress factors. It was suggested that the light-sensing pathways in C. Unicolor are cross-linked with stress signaling and secretion of proteases presumably serving as regulatory molecules.

  • Combined Effect of Light and Nutrients on the Micromorphology of the White rot Fungus Cerrena Unicolor.
    International journal of molecular sciences, 2020
    Co-Authors: Anna Pawlik, Jerzy Wielbo, Andrzej Mazur, Magdalena Jaszek, Dawid Stefaniuk, Piotr Koper, Kamil Żebracki, Urszula Świderska-burek, Grzegorz Janusz
    Abstract:

    Light influences developmental pathways in fungi. Recent transcriptomic and biochemical analyses have demonstrated that light influences the metabolism of a white-rot basidiomycete Cerrena Unicolor. However, the expression profile of genes involved in the growth and development, or micromorphological observations of the mycelium in response to variable lighting and culturing media, have not performed. We aim to reveal the effect of light and nutrients on C. Unicolor growth and a potential relationship between the culture medium and lighting conditions on fungus micromorphological structures. Confocal laser scanning microscopy and scanning electron microscopy were employed for morphological observations of C. Unicolor mycelium cultivated in red, blue, green, and white light and darkness on mineral and sawdust media. A comprehensive analysis of C. Unicolor differentially expressed genes (DEGs) was employed to find global changes in the expression profiles of genes putatively involved in light-dependent morphogenesis. Both light and nutrients influenced C. Unicolor growth and development. Considerable differences in the micromorphology of the mycelia were found, which were partially reflected in the functional groups of DEGs observed in the fungus transcriptomes. A complex cross-interaction of nutritional and environmental signals on C. Unicolor growth and morphology was suggested. The results are a promising starting point for further investigations of fungus photobiology.

  • Light-regulated synthesis of extra- and intracellular enzymes related to wood degradation by the white rot fungus Cerrena Unicolor during solid-state fermentation on ash sawdust-based medium.
    Acta biochimica Polonica, 2019
    Co-Authors: Anna Pawlik, Justyna Sulej, Magdalena Jaszek, Grzegorz Janusz
    Abstract:

    The light-dependent metabolism of the white rot basidiomycete Cerrena Unicolor FCL139 has already been demonstrated using transcriptomic and Biolog-based approaches. To further analyze the influence of light on C. Unicolor wood degradation, we measured the activity of an array of CAZymes (carbohydrate-active enzymes) and enzymes involved in the redox system of fungal cells associated with lignolysis. Extra- and intracellular enzymatic extracts were obtained from solid-state ash sawdust C. Unicolor cultures cultivated for 14 days under red, blue, green, or white light conditions, or in the dark. Light greatly influenced the synthesis of MnP, total cellulases, endo-1,4-β-glucanase, endo-1,4-β-xylanase, catalase, and superoxide dismutase. The production of MnP and catalase was evidently stimulated by white light. It is also worth noticing that blue light caused a gradual increase in the activity of total cellulases throughout the entire period of C. Unicolor growth. Moreover, endo-1,4-β-glucanase showed the highest activity on day 13 of fungus cultivation and the production of laccase and β-glucosidase appeared to be the least influenced by light. However, the strongest activity of the endo-1,4-β-xylanase was observed in the dark. It seemed that light not only influenced the regulation of the synthesis of the wood-degrading enzymes at different levels, but also acted indirectly by affecting production of enzymes managing harmful lignin by-products causing oxidative stress. The ability of the fungus to decompose woody plant material is clearly influenced by environmental factors.

  • The wood decay fungus Cerrena Unicolor adjusts its metabolism to grow on various types of wood and light conditions.
    PLOS ONE, 2019
    Co-Authors: Anna Pawlik, Marta Ruminowicz-stefaniuk, Jerzy Wielbo, Magdalena Frąc, Andrzej Mazur, Grzegorz Janusz
    Abstract:

    Cerrena Unicolor is a wood-degrading basidiomycete with ecological and biotechnological importance. Comprehensive Biolog-based analysis was performed to assess the metabolic capabilities and sensitivity to chemicals of C. Unicolor FCL139 growing in various sawdust substrates and light conditions. The metabolic preferences of the fungus towards utilization of specific substrates were shown to be correlated with the sawdust medium applied for fungus growth and the light conditions. The highest catabolic activity of C. Unicolor was observed after fungus precultivation on birch and ash sawdust media. The fungus growing in the dark showed the highest metabolic activity which was indicated by capacity to utilize a broad spectrum of compounds and the decomposition of 74/95 of the carbon sources. In all the culture light conditions, p-hydroxyphenylacetic acid was the most readily metabolized compound. The greatest tolerance to chemicals was also observed during C. Unicolor growth in darkness. The fungus was the most sensitive to nitrogen compounds and antibiotics, but more resistant to chelators. Comparative analysis of C. Unicolor and selected wood-decay fungi from different taxonomic and ecological groups revealed average catabolic activity of the fungus. However, C. Unicolor showed outstanding capabilities to catabolize salicin and arbutin. The obtained picture of C. Unicolor metabolism showed that the fungus abilities to decompose woody plant material are influenced by various environmental factors.

  • RNA Sequencing Reveals Differential Gene Expression of Cerrena Unicolor in Response to Variable Lighting Conditions.
    International journal of molecular sciences, 2019
    Co-Authors: Anna Pawlik, Jerzy Wielbo, Andrzej Mazur, Piotr Koper, Kamil Żebracki, Agnieszka Kubik-komar, Grzegorz Janusz
    Abstract:

    To elucidate the light-dependent gene expression in Cerrena Unicolor FCL139, the transcriptomes of the fungus growing in white, blue, green, and red lighting conditions and darkness were analysed. Among 10,413 all-unigenes detected in C. Unicolor, 7762 were found to be expressed in all tested conditions. Transcripts encoding putative fungal photoreceptors in the C. Unicolor transcriptome were identified. The number of transcripts uniquely produced by fungus ranged from 20 during its growth in darkness to 112 in the green lighting conditions. We identified numerous genes whose expression differed substantially between the darkness (control) and each of the light variants tested, with the greatest number of differentially expressed genes (DEGs) (454 up- and 457 down-regulated) observed for the white lighting conditions. The DEGs comprised those involved in primary carbohydrate metabolism, amino acid metabolism, autophagy, nucleotide repair systems, signalling pathways, and carotenoid metabolism as defined using Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The analysis of the expression profile of genes coding for lignocellulose-degrading enzymes suggests that the wood-degradation properties of C. Unicolor may be independent of the lighting conditions and may result from the overall stimulation of fungal metabolism by daylight.

Anna Jarosz-wilkołazka - One of the best experts on this subject based on the ideXlab platform.

  • New bioactive fungal molecules with high antioxidant and antimicrobial capacity isolated from Cerrena Unicolor idiophasic cultures.
    BioMed research international, 2013
    Co-Authors: Magdalena Jaszek, Justyna Sulej, Grzegorz Janusz, Anna Matuszewska, Monika Osinska-jaroszuk, Dawid Stefaniuk, Jolanta Polak, Marta Ruminowicz, Krzysztof Grzywnowicz, Anna Jarosz-wilkołazka
    Abstract:

    Three bioactive fractions, extracellular laccase (ex-LAC), crude endopolysaccharides (c-EPL), and a low molecular subfraction of secondary metabolites (ex-LMS), were isolated from the idiophasic cultures of the white rot fungus Cerrena Unicolor. For the first time, we determined the antioxidant properties of these samples by chemiluminometric measurement (a) and assessment of the scavenging effect on ABTS (b) and the DPPH reduction rate (c). The highest reducing capability was found for the ex-LMS fraction: 39–90% for (a), 20–90% for (b), and 10–59% for (c) at the concentration of 6.25–800 µg/mL. The scavenging abilities of the C. Unicolor c-EPL were between 36 and 70% for (a), 2 and 60% for (b), and 28 and 32% for (c) at the concentration of 6.25–800 µg/mL. A very high prooxidative potential was observed for the ex-LAC probes. The preliminary toxicity tests were done using the Microtox system and revealed the following percentage of the toxic effect against Vibrio fischeri: 85.37% for c-EPL, 50.67% for ex-LAC, and 99.8% for ex-LMS, respectively. The ex-LAC sample showed the antibacterial activity against Escherichia coli, c-EPL against Staphylococcus aureus, and ex-LMS against both bacterial strains, respectively, but the stronger inhibitory effect was exerted on S. aureus.

  • Synthesis and structural characterization of a novel phenoxazinone dye by use of a fungal laccase
    Journal of Molecular Catalysis B-enzymatic, 2010
    Co-Authors: Stefania Forte, Jolanta Polak, Anna Jarosz-wilkołazka, Daniela Valensin, Maurizio Taddei, Riccardo Basosi, Sophie Vanhulle, Rebecca Pogni
    Abstract:

    Laccase, isolated from Cerrena Unicolor, is able to transform 3-amino-4-hydroxybenzensulfonic acid into a water soluble phenoxazine dye with an extinction coefficient (epsilon) of 8600 M-1 cm(-1). The dye has been characterized using a variety of different analytic and spectroscopic techniques like UV-vis spectroscopy, HPLC (High Performance Liquid Chromatography), ESI/MS (Electrospray Ionization Mass Spectrometry) and the following NMR experiments: H-1, C-13, TOCSY (Total Correlation Spectroscopy), HSQC (Heteronuclear Single Quantum Coherence). HMBC (Heteronuclear Multiple Bond Coherence) showing the structure of 2-amino-3-oxo-3H-phenoxazine-8-sulfonic acid. The advantages of the presented biocatalytic system, in alignment with chemical system to obtain Curie_22, are eco-sustainability and one step performance. (C) 2010 Elsevier B.V. All rights reserved.

  • Catalytic activity of Cerrena Unicolor laccase in aqueous solutions of water-miscible organic solvents—Experimental and numerical description
    Journal of Molecular Catalysis B: Enzymatic, 2007
    Co-Authors: Janina Rodakiewicz-nowak, Anna Jarosz-wilkołazka
    Abstract:

    Abstract The stability and catalytic activity during oxidation of syringaldazine (SGZ) by laccase from Cerrena Unicolor (CUL) was studied in aqueous solutions of ethanol, acetone and dimethyl sulfoxide (DMSO). Additional, the oxidation of other two substrates, catechol (CAT) and 2,6-dimethoxyphenol (DMOP), was studied in ethanol and in DMSO solutions, respectively. It was observed that in 3 M solutions of the studied solvents CUL was practically stable for 30 min of the experiments, which show its good applicability in solutions of water-miscible organic solvents. Moreover, it was observed that the effect of the tested organic solvents on the maximum rate of oxidation values ( V max ) of different substrates was not dependent on the substrate. Within the studied range of concentrations (up to 8 M) ethanol did not affect V max values and the presence of DMSO reduced it at roughly the same rate for the studied substrates. In the case of SGZ as a substrate, the effect of the solvent on V max values increased in the order ethanol  K M in the order DMSO

  • Species-specific Cd-stress Response in the White Rot Basidiomycetes Abortiporus biennis and Cerrena Unicolor
    Biometals, 2006
    Co-Authors: Anna Jarosz-wilkołazka, Marcin Grąz, Bianca Braha, Sieglinde Menge, Dietmar Schlosser, Gerd-joachim Krauss
    Abstract:

    The effect of cadmium (Cd) on fungal growth, Cd bioaccumulation and biosorption, and on the formation of potential heavy metal response indicators such as thiols, oxalate, and laccase was investigated in the white rot fungi Cerrena Unicolor and Abortiporus biennis . Only the highest Cd concentration employed (200  μ M) inhibited growth of C. Unicolor , whereas already lower Cd concentrations caused decreasing mycelia dry weights in A. biennis . Cd biosorption onto the mycelial surface was the predominant Cd sequestration mechanism in C. Unicolor . Surface-bound and bioaccumulated Cd concentrations were essentially in the same range in A. biennis , leading to considerably higher intracellular Cd concentrations in A. biennis than in C. Unicolor . Oxalate and laccase were produced by both of the fungal strains and their extracellular levels were elevated upon Cd exposure. Oxalate concentrations and laccase titres were considerably higher in C. Unicolor than in A. biennis . Both fungi responded to increasing Cd concentrations by increasing intracellular amounts of thiol compounds (cysteine, γ -glutamylcysteine, glutathione in both its reduced and oxidized form) but Cd application increased the amounts of thiols to a higher extend in A. biennis . Taken together, these species-specific responses towards Cd suggest that C. Unicolor possesses a more efficient system than A. biennis to keep intracellular Cd concentrations low.

  • Characterization of graphite electrodes modified with laccases from Trametes hirsuta and Cerrena Unicolor and their use for flow injection amperometric determination of some phenolic compounds
    International Journal of Environmental Analytical Chemistry, 2005
    Co-Authors: Behzad Haghighi, Lo Gorton, Anna Jarosz-wilkołazka, Tautgirdas Ruzgas, Andrzej Leonowicz
    Abstract:

    Fungal laccases from Trametes hirsuta and Cerrena Unicolor were immobilized on spectroscopic graphite electrodes using physical absorption. The laccase-modified graphite electrodes were mounted as the working electrodes into a wall jet flow through an electrochemical cell and then used for flow injection amperometric determination of different phenolic compounds of relevance both for environmental and clinical analysis. At optimum conditions for determination of catechol serving as a model compound, the output response signals of the laccase-modified electrodes were recorded as the result of injections of 50 μL of solutions with different phenolic substrates into the carrier citrate buffer solution with a working potential of −50 mV vs. Ag|AgCl. Statistical aspects were applied and the Michaelis–Menten constants evaluated were correlated to the chemical structure of the investigated phenolic compounds.

Magdalena Jaszek - One of the best experts on this subject based on the ideXlab platform.

  • Chemopreventive activity of bioactive fungal fractions isolated from milk-supplemented cultures of Cerrena Unicolor and Pycnoporus sanguineus on colon cancer cells
    3 Biotech, 2021
    Co-Authors: Mateusz Pięt, Magdalena Jaszek, Dawid Stefaniuk, A. Zając, R. Paduch, M. Frant, A. Matuszewska, K. Grzywnowicz
    Abstract:

    The biochemical properties and anti-tumorigenic activity of Cerrena Unicolor (CU) and Pycnoporus sanguineus (PS) towards colon cancer cells and the effect of supplementation of the fungal culture medium with cow milk on these activities were examined. CU1-II and PS4-II exhibited anticancer properties through various mechanisms. The extracts at the 200 µg/mL concentration significantly decreased the viability of HT-29 and SW948 cells. They also exhibited pro-apoptotic properties towards the cancer cell lines (HT-29, LS 180, and SW948). Furthermore, culturing the studied fungi on milk-supplemented media may improve the pro-health properties of both milk and mushrooms. The extracts had a higher concentration of proteins, lower levels of free amino acids, and higher content of phenolic compounds than milk. They also exerted a free radical scavenging effect, which may be connected with the high activity of catalase and superoxide dismutase. The tested extracts exhibited anticancer activity: C. Unicolor grown on the medium without milk and P. sanguineus grown on the medium with milk. The CU1-II and PS4-II extracts exhibited the strongest anticancer properties; however, PS4-II exerted a milder effect on normal CCD 841 CoTr cells than CU1-II. CU3-II exerted the mildest effect among all extracts on both normal and cancer cells.

  • Lighting Conditions Influence the Dynamics of Protease Synthesis and Proteasomal Activity in the White Rot Fungus Cerrena Unicolor.
    Biomolecules, 2020
    Co-Authors: Anna Pawlik, Marta Ruminowicz-stefaniuk, Andrzej Mazur, Magdalena Jaszek, Anita Swatek, Beata Ciołek, Grzegorz Janusz
    Abstract:

    Recent transcriptomic and biochemical studies have revealed that light influences the global gene expression profile and metabolism of the white-rot fungus Cerrena Unicolor. Here, we aimed to reveal the involvement of proteases and ubiquitin-mediated proteolysis by the 26S proteasome in the response of this fungus to white, red, blue and green lighting conditions and darkness. The changes in the expression profile of C. Unicolor genes putatively engaged in proteolysis were found to be unique and specific to the applied wavelength of light. It was also demonstrated that the activity of proteases in the culture fluid and mycelium measured using natural and synthetic substrates was regulated by light and was substrate-dependent. A clear influence of light on protein turnover and the qualitative and quantitative changes in the hydrolytic degradation of proteins catalyzed by various types of proteases was shown. The analysis of activity associated with the 26S proteasome showed a key role of ATP-dependent proteolysis in the initial stages of adaptation of fungal cells to the stress factors. It was suggested that the light-sensing pathways in C. Unicolor are cross-linked with stress signaling and secretion of proteases presumably serving as regulatory molecules.

  • Combined Effect of Light and Nutrients on the Micromorphology of the White rot Fungus Cerrena Unicolor.
    International journal of molecular sciences, 2020
    Co-Authors: Anna Pawlik, Jerzy Wielbo, Andrzej Mazur, Magdalena Jaszek, Dawid Stefaniuk, Piotr Koper, Kamil Żebracki, Urszula Świderska-burek, Grzegorz Janusz
    Abstract:

    Light influences developmental pathways in fungi. Recent transcriptomic and biochemical analyses have demonstrated that light influences the metabolism of a white-rot basidiomycete Cerrena Unicolor. However, the expression profile of genes involved in the growth and development, or micromorphological observations of the mycelium in response to variable lighting and culturing media, have not performed. We aim to reveal the effect of light and nutrients on C. Unicolor growth and a potential relationship between the culture medium and lighting conditions on fungus micromorphological structures. Confocal laser scanning microscopy and scanning electron microscopy were employed for morphological observations of C. Unicolor mycelium cultivated in red, blue, green, and white light and darkness on mineral and sawdust media. A comprehensive analysis of C. Unicolor differentially expressed genes (DEGs) was employed to find global changes in the expression profiles of genes putatively involved in light-dependent morphogenesis. Both light and nutrients influenced C. Unicolor growth and development. Considerable differences in the micromorphology of the mycelia were found, which were partially reflected in the functional groups of DEGs observed in the fungus transcriptomes. A complex cross-interaction of nutritional and environmental signals on C. Unicolor growth and morphology was suggested. The results are a promising starting point for further investigations of fungus photobiology.

  • Light-regulated synthesis of extra- and intracellular enzymes related to wood degradation by the white rot fungus Cerrena Unicolor during solid-state fermentation on ash sawdust-based medium.
    Acta biochimica Polonica, 2019
    Co-Authors: Anna Pawlik, Justyna Sulej, Magdalena Jaszek, Grzegorz Janusz
    Abstract:

    The light-dependent metabolism of the white rot basidiomycete Cerrena Unicolor FCL139 has already been demonstrated using transcriptomic and Biolog-based approaches. To further analyze the influence of light on C. Unicolor wood degradation, we measured the activity of an array of CAZymes (carbohydrate-active enzymes) and enzymes involved in the redox system of fungal cells associated with lignolysis. Extra- and intracellular enzymatic extracts were obtained from solid-state ash sawdust C. Unicolor cultures cultivated for 14 days under red, blue, green, or white light conditions, or in the dark. Light greatly influenced the synthesis of MnP, total cellulases, endo-1,4-β-glucanase, endo-1,4-β-xylanase, catalase, and superoxide dismutase. The production of MnP and catalase was evidently stimulated by white light. It is also worth noticing that blue light caused a gradual increase in the activity of total cellulases throughout the entire period of C. Unicolor growth. Moreover, endo-1,4-β-glucanase showed the highest activity on day 13 of fungus cultivation and the production of laccase and β-glucosidase appeared to be the least influenced by light. However, the strongest activity of the endo-1,4-β-xylanase was observed in the dark. It seemed that light not only influenced the regulation of the synthesis of the wood-degrading enzymes at different levels, but also acted indirectly by affecting production of enzymes managing harmful lignin by-products causing oxidative stress. The ability of the fungus to decompose woody plant material is clearly influenced by environmental factors.

  • Antitumor potential of new low molecular weight antioxidative preparations from the white rot fungus Cerrena Unicolor against human colon cancer cells
    Scientific reports, 2019
    Co-Authors: Anna Matuszewska, Magdalena Jaszek, Dawid Stefaniuk, Marcin Grąz, Mateusz Pięt, Adrian Zając, Łukasz Matuszewski, Iga Cios, Roman Paduch, Renata Bancerz
    Abstract:

    The aim of this study was to investigate the anticancer and antioxidant activities of low molecular weight subfractions isolated from secondary metabolites produced by the wood degrading fungus Cerrena Unicolor. Human colon cancer cells (stage I) HT-29 and human normal colon epithelial cells CCD 841 CoTr were used in the research. The present study demonstrated that the low molecular weight subfractions exhibited inhibitory activity towards human colon cancer cells HT-29 at a concentration range of 25–200 μg/mL. All 6 subfractions inhibited proliferation of cells down to 47.5–9.2% at the highest concentrations in a dose-dependent manner. The most desired activity was exhibited by subfractions S, 3, 4, and 5, as the proliferation of HT-29 cells was inhibited to the greatest extent (16.5, 47.5, 42.7, and 26.1% of the control, respectively), while the effect on CCD 841 CoTr cells was the mildest (inhibition to 54.4, 71.4, 79.4, and 53.4%, compared to the control, respectively). The microscopic observation revealed that all extracts induced programmed cell death, i.e. apoptosis (up to 44.4% (subfraction 6) towards HT-29 and less than 20% (most fractions) towards CCD 841 CoTr), with no or a significantly low level of necrosis in both cell lines at the same time.

Jerzy Rogalski - One of the best experts on this subject based on the ideXlab platform.

  • characterization of cellobiose dehydrogenase from a biotechnologically important Cerrena Unicolor strain
    Applied Biochemistry and Biotechnology, 2015
    Co-Authors: Justyna Sulej, Monika Osinskajaroszuk, Iwona Komaniecka, Adam Choma, Patrycja Rachubik, Grzegorz Janusz, Andrzej Mazur, Jerzy Rogalski
    Abstract:

    Cellobiose dehydrogenase (CDH), a secreted flavocytochrome produced by a number of wood-degrading fungi, was detected in the culture supernatant of a biotechnologically important strain of Cerrena Unicolor grown in a modified cellulose-based liquid medium. The enzyme was purified as two active fractions: CuCDH-FAD (flavin domain) (1.51-fold) with recovery of 8.35 % and CuCDH (flavo-heme enzyme) (21.21-fold) with recovery of 73.41 %. As CDH from other wood-rotting fungi, the intact form of cellobiose dehydrogenase of C. Unicolor is a monomeric protein containing one flavin and one heme b with molecular mass 97 kDa and pI = 4.55. The enzyme is glycosylated (8.2 %) mainly with mannose and glucosamine residues. Moreover, the cellobiose dehydrogenase gene cdh1 and its corresponding cDNA from the fungus C. Unicolor were isolated, cloned, and characterized. The 2316-bp full-length cDNA of cdh1 encoded a mature CDH protein containing 771 amino acids preceded by a signal peptide consisting of 18 amino acids. Moreover, both active fractions were characterized in terms of kinetics, temperature and pH optima, and antioxidant properties.

  • Characterization of Cellobiose Dehydrogenase from a Biotechnologically Important Cerrena Unicolor
    2015
    Co-Authors: Adam Choma, Jerzy Rogalski
    Abstract:

    # The Author(s) 2015. This article is published with open access at Springerlink.com Abstract Cellobiose dehydrogenase (CDH), a secreted flavocytochrome produced by a num-ber of wood-degrading fungi, was detected in the culture supernatant of a biotechnologically important strain of Cerrena Unicolor grown in a modified cellulose-based liquid medium. The enzyme was purified as two active fractions: CuCDH-FAD (flavin domain) (1.51-fold) with recovery of 8.35 % and CuCDH (flavo-heme enzyme) (21.21-fold) with recovery of 73.41 %. As CDH from other wood-rotting fungi, the intact form of cellobiose dehydrogenase of C. Unicolor is a monomeric protein containing one flavin and one heme b with molecular mass 97 kDa and pI=4.55. The enzyme is glycosylated (8.2 %) mainly with mannose and glucos-amine residues. Moreover, the cellobiose dehydrogenase gene cdh1 and its corresponding cDNA from the fungus C. Unicolor were isolated, cloned, and characterized. The 2316-bp full-length cDNA of cdh1 encoded a mature CDH protein containing 771 amino acids preceded by a signal peptide consisting of 18 amino acids. Moreover, both active fractions were characterized in terms of kinetics, temperature and pH optima, and antioxidant properties

  • Chemical Modifications of Laccase from White-Rot Basidiomycete Cerrena Unicolor
    Applied biochemistry and biotechnology, 2012
    Co-Authors: Katarzyna H. Kucharzyk, Grzegorz Janusz, I. Karczmarczyk, Jerzy Rogalski
    Abstract:

    Laccases belong to the group of phenol oxidizes and constitute one of the most promising classes of enzymes for future use in various fields. For industrial and biotechnological purposes, laccases were among the first enzymes providing larger-scale applications such as removal of polyphenols or conversion of toxic compounds. The wood-degrading basidiomycete Cerrena Unicolor C-139, reported in this study, is one of the high-laccase producers. In order to facilitate novel and more efficient biocatalytic process applications, there is a need for laccases with improved biochemical properties, such as thermostability or stability in broad ranges of pH. In this work, modifications of laccase isoforms by hydrophobization, hydrophilization, and polymerization were performed. The hydrophobized and hydrophilized enzyme showed enhanced surface activity and higher ranges of pH and temperatures in comparison to its native form. However, performed modifications did not appear to noticeably alter enzyme’s native structure possibly due to the formation of coating by particles of saccharides around the molecule. Additionally, surface charge of modified laccase shifted towards the negative charge for the hydrophobized laccase forms. In all tested modifications, the size exclusion method led to average 80 % inhibition removal for hydrophilized samples after an hour of incubation with fluoride ions. Samples that were hydrophilized with lactose and cellobiose showed an additional 90 % reversibility of inhibition by fluoride ions after an hour of concluding the reaction and 40 % after 24 h. The hydrophobized laccase showed higher level of the reversibility after 1 h (above 80 %) and 24 h (above 70 %) incubation with fluoride ions. The addition of ascorbate to laccase solution before a fluoride spike resulted in more efficient reversibility of fluoride inhibitory effect in comparison to the treatments with reagents used in the reversed sequence.

  • PURIFICATION OF EXTRACELLULAR LACCASE FROM Cerrena Unicolor
    Preparative biochemistry & biotechnology, 2010
    Co-Authors: Jerzy Rogalski, Grzegorz Janusz
    Abstract:

    Cerrena Unicolor was found to produce large amounts of extracellular laccase when grown aerobically on the optimized Lindenberg and Holm medium in fermenter culture with an automatic pH control. The laccase from this source was purified to homogeneity by a rapid procedure, using ion-exchange chromatography, affinity chromatography, and chromatofocusing. The enzymes isoforms were recovered with a 65- to 92-fold increase in specific activity and a yield for Ia1 = 6.7%; Ia2 = 27.5%; Ib = 9.7%; and IIa1 = 21%. The molecular mass of the purified enzymes proved to be 45, 47, 54, and 62 kD, respectively, as determined by size-exclusion high-performance liquid chromatography (HPLC). The isoelectric points were in the range of 4.7 to 4.2, and the carbohydrate content in the purified enzymes was between 1.6 and 3.5%.

  • Pyrene sulfonate functionalised single-walled carbon nanotubes for mediatorless dioxygen bioelectrocatalysis
    Electrochemistry Communications, 2009
    Co-Authors: Martin Jönsson-niedziolka, Jerzy Rogalski, Katarzyna Szot, Marcin Opallo
    Abstract:

    Abstract Using one step procedure we have prepared electrodes using single-walled carbon nanotubes non-covalently functionalised with 1,3,6,8-pyrenetetrasulfonate or 1-pyrenesulfonate immobilised together with the enzyme laccase (from Cerrena Unicolor ) in a silicate sol–gel matrix. Both electrodes exhibit mediatorless electrocatalytic activity towards dioxygen reduction with a current density of more than 100 μA cm −2 . It is more than two times higher than obtained with analogous electrode modified with raw single-walled carbon nanotubes which is also mechanically unstable.

Anli Geng - One of the best experts on this subject based on the ideXlab platform.

  • Expression of a novel manganese peroxidase from Cerrena Unicolor BBP6 in Pichia pastoris and its application in dye decolorization and PAH degradation
    Biochemical Engineering Journal, 2020
    Co-Authors: Hao Zhang, Xiaoyu Zhang, Anli Geng
    Abstract:

    Abstract Manganese peroxidase (MnP), which is an extracellular glycosylated heme protein produced by various basidiomycetous fungi, shows attractive potential in various biotechnological applications. A novel MnP gene, mnp3, from white-rot fungus Cerrena Unicolor BBP6 was cloned and expressed in Pichia pastoris with a MnP activity of 154.5 U/l. The recombinant enzyme, rMnP3-BBP6, had strong decolorizing activity on a variety of dyes and it was efficient in denim bleaching. In addition, it was able to degrade fluorene and phenanthrene effectively. The results of this research demonstrated that rMnP3-BBP6 had promising biotechnological application potential in textile industries and polycyclic aromatic hydrocarbon bioremediation.

  • Directed Evolution of a Homodimeric Laccase from Cerrena Unicolor BBP6 by Random Mutagenesis and In Vivo Assembly
    International journal of molecular sciences, 2018
    Co-Authors: Ji Zhang, Xiaoyu Zhang, Anli Geng
    Abstract:

    Laccases have great potential for industrial applications due to their green catalytic properties and broad substrate specificities, and various studies have attempted to improve the catalytic performance of these enzymes. Here, to the best of our knowledge, we firstly report the directed evolution of a homodimeric laccase from Cerrena Unicolor BBP6 fused with α-factor prepro-leader that was engineered through random mutagenesis followed by in vivo assembly in Saccharomyces cerevisiae. Three evolved fusion variants selected from ~3500 clones presented 31- to 37-fold increases in total laccase activity, with better thermostability and broader pH profiles. The evolved α-factor prepro-leader enhanced laccase expression levels by up to 2.4-fold. Protein model analysis of these variants reveals that the beneficial mutations have influences on protein pKa shift, subunit interaction, substrate entrance, and C-terminal function.

  • A novel homodimer laccase from Cerrena Unicolor BBP6: Purification, characterization, and potential in dye decolorization and denim bleaching.
    PloS one, 2018
    Co-Authors: Ji Zhang, Xiaoyu Zhang, Hao Zhang, Lei Sun, Shufang Wang, Anli Geng
    Abstract:

    The white-rot fungus Cerrena Unicolor BBP6 produced up to 243.4 U mL-1 laccase. A novel laccase isoform LacA was purified; LacA is a homodimer with an apparent molecular mass of 55 kDa and an isoelectric point of 4.7. Its optimal pH was 2.5, 4.0, and 5.5 when 2, 2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), guaiacol, and 2, 6-dimethoxyphenol (2, 6-DMP) were used as the substrates, respectively. The optimal temperature was 60°C for ABTS and 80°C for both guaiacol and 2, 6-DMP. LacA retained 82-92% activity when pH was greater than 4 and 42%-92% activity at or below 50°C. LacA was completely inhibited by 0.1 mM L-cysteine, 1 mM Dithiothreitol, and 10 mM metal ions, Ca2+, Mg2+ and Co2+. LacA had good affinity for ABTS, with a Km of 49.1 μM and a kcat of 3078.9 s-1. It decolorized synthetic dyes at 32.3-87.1%. In the presence of 1-hydroxybenzotriazole (HBT), LacA decolorized recalcitrant dyes such as Safranine (97.1%), Methylene Blue (98.9%), Azure Blue (96.6%) and simulated textile effluent (84.6%). With supplemented manganese peroxidase (MnP), Mn2+ and HBT, the purified LacA and BBP6 fermentation broth showed great potential in denim bleaching, with an up to 5-fold increase in reflectance values.

  • Purification and characterization of a novel manganese peroxidase from white-rot fungus Cerrena Unicolor BBP6 and its application in dye decolorization and denim bleaching
    Process Biochemistry, 2018
    Co-Authors: Hao Zhang, Ji Zhang, Xiaoyu Zhang, Anli Geng
    Abstract:

    Abstract Manganese peroxidase, with molecular mass of 45 kDa, was purified from the white-rot fungus Cerrena Unicolor BBP6 and named MnP-BBP6. The optimum temperature and pH of MnP-BBP6 activity were 60 °C and 4.5, respectively. MnP-BBP6 showed high stability toward many metal ions. It could effectively decolorize many types of dyes including Congo red (53.9% in 12 h), methyl orange (77.6% in 12 h), Remazol brilliant blue R (81.0% in 5 h), bromophenol blue (62.2% in 12 h) and crystal violet (80.9% in 12 h). With gallic acid as the redox mediator, azure blue was decolorized by 63.1% in 24 h. In addition to dye decolorization, purified MnP-BBP6 also presented significant effect on denim bleaching, with up to 3-fold increase in reflectance without mediators. With such strong decolorizing activity, MnP-BBP6 was demonstrated to be a potential peroxidase in the textile industry, and its decolorization potential was greatly enhanced with the addition of redox mediators.