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.

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 Effect of different wavelengths of light on laccase, cellobiose dehydrogenase, and proteases produced by Cerrena unicolor, Pycnoporus sanguineus and Phlebia lindtneri.

2016 ◽  
Vol 63 (2) ◽  
Author(s):  
Grzegorz Janusz ◽  
Justyna Sulej ◽  
Magdalena Jaszek ◽  
Monika Osińska-Jaroszuk
Keyword(s):  
Blue Light ◽  
White Light ◽  
Green Light ◽  
White Rot Fungi ◽  
Cellobiose Dehydrogenase ◽  
White Rot ◽  
Pycnoporus Sanguineus ◽  
Lighting Conditions ◽  
Cerrena Unicolor ◽  
The Media

Three species of white rot fungi: Cerrena unicolor, Phlebia lindtneri and Pycnoporus sanguineus were cultured in two different media under five different lighting conditions: dark, white, red, blue, and green light. Laccase, cellobiose dehydrogenase, and protease activities were examined in the samples. Blue light efficiently boosted laccase synthesis in C. unicolor and P. sanguineus, whereas the highest activities (20 654 nkat/l) of P. lindtneri laccase were observed when this fungus was maintained in green light. On the contrary, the green light allowed obtaining the highest activities of cellobiose dehydrogenase of C. unicolor and P. lindtneri, while CDH of P. sanguineus seems to be dependent on white light. It is clearly visible that differences in protease activities are noticeable not only between the lights variants but also among the media used. However, high proteases activities are correlated with light variants inducing laccase in Lindeberg and Holm medium. Contrary to the cellulose-based medium, where they are weak in light variants that lead to high CDH activities.

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.

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.

scholarly journals 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

2019 ◽  
Author(s):  
Anna Pawlik ◽  
Magdalena Jaszek ◽  
Justyna Sulej ◽  
Grzegorz Janusz
Keyword(s):  
Solid State ◽  
White Light ◽  
Redox System ◽  
White Rot ◽  
White Rot Fungus ◽  
Wood Degradation ◽  
Dark Light ◽  
Cerrena Unicolor ◽  
By Products ◽  
Different Levels

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.

scholarly journals Genomic Studies of White-Rot Fungus Cerrena unicolor SP02 Provide Insights into Food Safety Value-Added Utilization of Non-Food Lignocellulosic Biomass

2021 ◽  
Vol 7 (10) ◽  
pp. 835
Author(s):  
Zichen Zhang ◽  
Aabid Manzoor Shah ◽  
Hassan Mohamed ◽  
Yao Zhang ◽  
Nino Tsiklauri ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Molecular Mechanisms ◽  
Genetic Material ◽  
Value Added ◽  
White Rot ◽  
White Rot Fungus ◽  
Industrial Biotechnology ◽  
Lignocellulolytic Enzymes ◽  
Cerrena Unicolor ◽  
Wide Range

Cerrena unicolor is an ecologically and biotechnologically important wood-degrading basidiomycete with high lignocellulose degrading ability. Biological and genetic investigations are limited in the Cerrena genus and, thus, hinder genetic modification and commercial use. The aim of the present study was to provide a global understanding through genomic and experimental research about lignocellulosic biomass utilization by Cerrena unicolor. In this study, we reported the genome sequence of C. unicolor SP02 by using the Illumina and PacBio 20 platforms to obtain trustworthy assembly and annotation. This is the combinational 2nd and 3rd genome sequencing and assembly of C. unicolor species. The generated genome was 42.79 Mb in size with an N50 contig size of 2.48 Mb, a G + C content of 47.43%, and encoding of 12,277 predicted genes. The genes encoding various lignocellulolytic enzymes including laccase, lignin peroxidase, manganese peroxidase, cytochromes P450, cellulase, xylanase, α-amylase, and pectinase involved in the degradation of lignin, cellulose, xylan, starch, pectin, and chitin that showed the C. unicolor SP02 potentially have a wide range of applications in lignocellulosic biomass conversion. Genome-scale metabolic analysis opened up a valuable resource for a better understanding of carbohydrate-active enzymes (CAZymes) and oxidoreductases that provide insights into the genetic basis and molecular mechanisms for lignocellulosic degradation. The C. unicolor SP02 model can be used for the development of efficient microbial cell factories in lignocellulosic industries. The understanding of the genetic material of C. unicolor SP02 coding for the lignocellulolytic enzymes will significantly benefit us in genetic manipulation, site-directed mutagenesis, and industrial biotechnology.

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