scholarly journals Cerrena unicolor Laccases, Genes Expression and Regulation of Activity

Biomolecules ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 468
Author(s):  
Anna Pawlik ◽  
Beata Ciołek ◽  
Justyna Sulej ◽  
Andrzej Mazur ◽  
Przemysław Grela ◽  
...  
Keyword(s):  
Laccase Activity ◽  
Environmental Changes ◽  
Specific Activity ◽  
Electrochemical Analysis ◽  
White Rot ◽  
White Rot Fungus ◽  
Cerrena Unicolor ◽  
Genes Expression ◽  
Regulation Of Activity ◽  
Laccase Genes

A white rot fungus Cerrena unicolor has been identified as an important source of laccase, unfortunately regulation of this enzyme genes expression is poorly understood. Using 1D and 2D PAGE and LC-MS/MS, laccase isoenzymes were investigated in the liquid filtrate of C. unicolor culture. The level of expression of laccase genes was measured using qPCR. The elevated concentrations of copper and manganese in the medium caused greatest change in genes expression and three laccase transcripts were significantly affected after culture temperature was decreased from 28 to 4 °C or increased to 40 °C. The small differences in the PAGE band intensities of individual laccase proteins were also observed, indicating that given compound affect particular laccase’s transcript. Analyses of laccase-specific activity, at all tested conditions, showed the increased activities as compared to the control, suggesting that enzyme is regulated at the post-translational stage. We observed that the aspartic protease purified from C. unicolor, significantly stimulate laccase activity. Moreover, electrochemical analysis of protease-treated laccase sample had 5 times higher redox peaks. The obtained results indicate that laccases released by C. unicolor are regulated at transcriptional, translational, and at the post-translational steps of gene expression helping fungus adapt to the environmental changes.

scholarly journals Improvement of laccase activity by silencing PacC in Ganoderma lucidum

2021 ◽  
Author(s):  
Mingwen Zhao ◽  
Jing Zhu ◽  
Shuqi Song ◽  
Lindan Lian ◽  
Liang Shi ◽  
...  
Keyword(s):  
Ganoderma Lucidum ◽  
Laccase Activity ◽  
Ph Value ◽  
Fold Increase ◽  
White Rot ◽  
White Rot Fungus ◽  
Copper Binding ◽  
Multicopper Oxidases ◽  
Binding Domains ◽  
Laccase Genes

Abstract Ganoderma lucidum is a representative white-rot fungus that has great potential to degrade lignocellulose biomass. Laccase is recognized as a class of the most important lignin-degrading enzymes in G. lucidum. However, the comprehensive regulatory mechanisms of laccase are still lacking. Based on the genome sequence of G. lucidum, 15 laccase genes were identified and their encoding proteins were analyzed in this study. All of the laccase proteins are predicted to be multicopper oxidases with conserved copper-binding domains. Most laccase proteins were secreted enzymes in addition to Lac14 in which the signal peptide could not be predicted. The activity of all laccases showed the highest level at pH 3.0 or pH 7.0, with total laccase activity of approximately 200 U/mg protein. Silencing PacC resulted in a 5.2 fold increase in laccase activity compared with WT. Five laccase genes (lac1, lac6, lac9, lac10 and lac14) showed an increased transcription levels (approximately 1.5-5.6 fold) in the PacC-silenced strains versus that in WT, while other laccase genes were downregulated or unchanged. The extracellular pH value was about 3.1, which was more acidic in the PacC-silenced strains than in the WT (pH 3.5). Moreover, maintaining the fermentation pH resulted in a downregulation of laccase activity which is induced by silencing PacC Our findings indicate that in addition to its function in acidification of environmental pH, PacC plays an important role in regulating laccase activity in fungi.

scholarly journals Introducing a Thermo-Alkali-Stable, Metallic Ion-Tolerant Laccase Purified From White Rot Fungus Trametes hirsuta

2021 ◽  
Vol 12 ◽  
Author(s):  
Jing Si ◽  
Hongfei Ma ◽  
Yongjia Cao ◽  
Baokai Cui ◽  
Yucheng Dai
Keyword(s):  
Specific Activity ◽  
Endocrine Disrupting Chemicals ◽  
Environmental Pollutants ◽  
Fold Increase ◽  
Industrial Applications ◽  
White Rot ◽  
White Rot Fungus ◽  
Metallic Ions ◽  
Trametes Hirsuta ◽  
Ph Range

This study introduces a valuable laccase, designated ThLacc-S, purified from white rot fungus Trametes hirsuta. ThLacc-S is a monomeric protein in nature with a molecular weight of 57.0 kDa and can efficiently metabolize endocrine disrupting chemicals. The enzyme was successfully purified to homogeneity via three consecutive steps consisting of salt precipitation and column chromatography, resulting in a 20.76-fold increase in purity and 46.79% yield, with specific activity of 22.111 U/mg protein. ThLacc-S was deciphered as a novel member of the laccase family and is a rare metalloenzyme that contains cysteine, serine, histidine, and tyrosine residues in its catalytic site, and follows Michaelis-Menten kinetic behavior with a Km and a kcat/Km of 87.466 μM and 1.479 s–1μM–1, respectively. ThLacc-S exerted excellent thermo-alkali stability, since it was markedly active after a 2-h incubation at temperatures ranging from 20 to 70°C and retained more than 50% of its activity after incubation for 72 h in a broad pH range of 5.0–10.0. Enzymatic activities of ThLacc-S were enhanced and preserved when exposed to metallic ions, surfactants, and organic solvents, rendering this novel enzyme of interest as a green catalyst for versatile biotechnological and industrial applications that require these singularities of laccases, particularly biodegradation and bioremediation of environmental pollutants.

Enhanced Formation of Extracellular Laccase Activity by the White-Rot Fungus Trametes multicolor

2002 ◽  
pp. 229-241 ◽  
Author(s):  
Johann Hess ◽  
Christian Leitner ◽  
Christiane Galhaup ◽  
Klaus D. Kulbe ◽  
Barbara Hinterstoisser ◽  
...  
Keyword(s):  
Laccase Activity ◽  
White Rot ◽  
White Rot Fungus ◽  
Extracellular Laccase ◽  
Trametes Multicolor

scholarly journals Lighting Conditions Influence the Dynamics of Protease Synthesis and Proteasomal Activity in the White Rot Fungus Cerrena unicolor

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1322
Author(s):  
Anna Pawlik ◽  
Magdalena Jaszek ◽  
Anita Swatek ◽  
Marta Ruminowicz-Stefaniuk ◽  
Beata Ciołek ◽  
...  
Keyword(s):  
Expression Profile ◽  
Hydrolytic Degradation ◽  
Culture Fluid ◽  
Global Gene Expression ◽  
26S Proteasome ◽  
Stress Factors ◽  
White Rot ◽  
White Rot Fungus ◽  
Lighting Conditions ◽  
Cerrena Unicolor

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.

Enhancement of lignin degradation and laccase activity in Pleurotus ostreatus by cotton stalk extract

1998 ◽  
Vol 44 (7) ◽  
pp. 676-680 ◽  
Author(s):  
Orly Ardon ◽  
Zohar Kerem ◽  
Yitzhak Hadar
Keyword(s):  
Oxygen Consumption ◽  
Pleurotus Ostreatus ◽  
Lignin Degradation ◽  
Laccase Activity ◽  
White Rot ◽  
White Rot Fungus ◽  
Lignin Biodegradation ◽  
Cotton Stalk ◽  
Fermentation System ◽  
Uv Visible

The white rot fungus Pleurotus ostreatus was grown in a chemically defined solid state fermentation system amended with cotton stalk extract (CSE).Treated cultures exhibited increased laccase activity as well as enhanced lignin mineralization. Mineralization of [14C]lignin initialized 4 days earlier in CSE-supplemented cultures than in control cultures. Total mineralization in the first 16 days was 15% in the CSE-treated cultures, compared with only 7% in the controls. Cotton stalk extract also contained compounds that serve as substrates for laccase purified from P. ostreatus as shown by oxygen consumption, as well as changes in the UV–visible spectrum.Key words: cotton, Pleurotusostreatus, white rot, laccase, lignin biodegradation.

scholarly journals A New Laccase of Lac 2 from the White Rot Fungus Cerrena unicolor 6884 and Lac 2-Mediated Degradation of Aflatoxin B1

Toxins ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 476 ◽  
Author(s):  
Zhimin Zhou ◽  
Renkuan Li ◽  
Tzi Bun Ng ◽  
Yunyun Lai ◽  
Jie Yang ◽  
...  
Keyword(s):  
Aflatoxin B1 ◽  
Environmental Pollutants ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Mass Spectrometry Data ◽  
White Rot ◽  
White Rot Fungus ◽  
Multicopper Oxidases ◽  
Cerrena Unicolor

Aflatoxin B1 (AFB1) is a known toxic human carcinogen and can be detoxified by laccases, which are multicopper oxidases that convert several environmental pollutants and toxins. In this study, a new laccase that could catalyze AFB1 degradation was purified and identified from the white-rot fungus Cerrena unicolor 6884. The laccase was purified using (NH4)2SO4 precipitation and anion exchange chromatography, and then identified as Lac 2 through zymogram and UHPLC-MS/MS based on the Illumina transcriptome analysis of C. unicolor 6884. Six putative laccase protein sequences were obtained via functional annotation. The lac 2 cDNA encoding a full-length protein of 512 amino acids was cloned and sequenced to expand the fungus laccase gene library for AFB1 detoxification. AFB1 degradation by Lac 2 was conducted in vitro at pH 7.0 and 45 °C for 24 h. The half-life of AFB1 degradation catalyzed by Lac 2 was 5.16 h. Acetosyringone (AS), Syrinagaldehyde (SA) and [2,2′ -azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)] (ABTS) at 1 mM concentration seemed to be similar mediators for strongly enhancing AFB1 degradation by Lac 2. The product of AFB1 degradation catalyzed by Lac 2 was traced and identified to be Aflatoxin Q1 (AFQ1) based on mass spectrometry data. These findings are promising for a possible application of Lac 2 as a new aflatoxin oxidase in degrading AFB1 present in food and feeds.

scholarly journals Combined Effect of Light and Nutrients on the Micromorphology of the White rot Fungus Cerrena unicolor

2020 ◽  
Vol 21 (5) ◽  
pp. 1678 ◽  
Author(s):  
Anna Pawlik ◽  
Magdalena Jaszek ◽  
Dawid Stefaniuk ◽  
Urszula Świderska-Burek ◽  
Andrzej Mazur ◽  
...  
Keyword(s):  
Growth And Development ◽  
Expression Profiles ◽  
Laser Scanning Microscopy ◽  
White Rot ◽  
White Rot Fungus ◽  
Confocal Laser ◽  
Global Changes ◽  
Cerrena Unicolor ◽  
Starting Point ◽  
Effect Of Light

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.

Induction of laccase activity in the white rot fungus Pleurotus ostreatus using water polluted with wheat straw extracts

2013 ◽  
Vol 133 ◽  
pp. 142-149 ◽  
Author(s):  
Alejandra Parenti ◽  
Elaia Muguerza ◽  
Amaia Redin Iroz ◽  
Alejandra Omarini ◽  
Enma Conde ◽  
...  
Keyword(s):  
Wheat Straw ◽  
Pleurotus Ostreatus ◽  
Laccase Activity ◽  
White Rot ◽  
White Rot Fungus

scholarly journals Effective and complex stimulation of the biodegradation system of fungus Cerrena unicolor by rapeseed meal fermentation

2016 ◽  
Vol 63 (3) ◽  
Author(s):  
Magdalena Jaszek ◽  
Justyna Miłek ◽  
Jerzy Żuchowski ◽  
Dawid Stefaniuk ◽  
Monika Prendecka
Keyword(s):  
Phenolic Compounds ◽  
Rapeseed Meal ◽  
White Rot ◽  
White Rot Fungus ◽  
Submerged Cultures ◽  
Biotechnological Production ◽  
Cerrena Unicolor ◽  
Zymographic Analysis ◽  
Stimulation Of ◽  
Antioxidative Ability

The effect of supplementation of medium with rapeseed meal (RM) on production of biotechnologically important enzymes was investigated in submerged cultures of the white rot fungus Cerrena unicolor. The addition of RM (3.5% w/v) distinctly stimulated the activities of laccase, chitinase, and β-glucosidase. As compared to the control, the activities of chitinase, β-glucosidase, and laccase in the RM supplemented cultures were up to 4.1, 8.4, and 3.9 times higher, respectively. The results of the spectrophotometric and spectrofluorometric measurements were additionally confirmed by zymographic analysis of the samples. The level of sugars and phenolic compounds as well as the antioxidative ability of fungal preparations were also determined. The results obtained indicate that the submerged liquid fermentation of rapeseed meal can be proposed as an inexpensive and very effective method for biotechnological production of chitinase, β-glucosidase, and laccase by C. unicolor.

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