Biodegradation of low-density polyethylene by mixed fungi composed of Alternaria sp. and Trametes sp. isolated from landfill sites.

With the development of industry and modern manufacturing, nondegradable low-density polyethylene (LDPE) has been widely used, posing a rising environmental hazard to natural ecosystems and public health. In this study, we isolated a series of LDPE-degrading fungi from landfill sites and carried out LDPE degradation experiments by combining highly efficient degrading fungi in pairs. The results showed that the mixed microorganisms composed of Alternaria sp. CPEF-1 and Trametes sp. PE2F-4 (H-3 group) had a greater degradation effect on heat-treated LDPE (T-LDPE). After 30 days of inoculation with combination strain H-3, the weight loss rate of the T-LDPE film was approximately 154% higher than that of the untreated LDPE (U-LDPE) film, and the weight loss rate reached 0.66 ± 0.06%. Environmental scanning electron microscopy (ESEM) and Fourier transform infrared spectroscopy (FTIR) were used to further investigate the biodegradation impacts of T-LDPE, including the changes on the surface and depolymerization of the LDPE films during the fungal degradation process. Our findings revealed that the combined fungal treatment is more effective at degrading T-LDPE than the single strain treatment, and it is expected that properly altering the composition of the microbial community can help lessen the detrimental impact of plastics on the environment.

Investigation of laccase activity in cholinium-based ionic liquids using experimental and molecular dynamics techniques.

Laccases hold great potential for biotechnological applications, particularly in environmental pollutant remediation. Laccase activity is governed by the solvent environment, and ionic liquids (ILs) emerge as a versatile solvent for activation or stabilization of enzymes. Herein, effects of cholinium-based ILs formulated with carboxylic acids, inorganic acid, and amino acids as anionic species, on the catalytic activity of laccase from Trametes versicolor were investigated by experimental and computational approaches. Experimental results showed that laccase activity was enhanced by 21.39 % in 0.5 M cholinium dihydrogen citrate ([Cho][DHC]), in relation to the laccase activity in phosphate buffer medium. However, cholinium aminoate ILs negatively affected laccase activity, as evidenced by the partial deactivation of laccase in both cholinium glycinate and cholinium phenylalaninate, at concentrations of 0.1 M and 0.5 M, respectively. Molecular dynamics studies revealed that the enhancement of laccase activity in [Cho][DHC] might be attributed to the highly stabilized and compact structure of laccase, facilitating a better internal electron transfer during the laccase-substrate interactions. Enhanced catalytic performance of laccase in [Cho][DHC] was postulated to be driven by the high accumulation level of dihydrogen citrate anions around laccase's surface. [Cho][DHC] holds great promise as a cosolvent in laccase-catalyzed biochemical reactions.

Impact of surface-active ionic solutions on the structure and function of laccase from trametes versicolor: Insights from molecular dynamics simulations.

Many protein-ionic liquid investigations have examined laccase interactions. Laccases are a class of poly-copper oxidoreductases that retain significant biotechnological relevance owing to their notable oxidative capabilities and their application in the elimination of synthetic dyes, phenolic compounds, insecticides, and various other substances. This study investigates the impact of surface active ionic liquids (SAILs), namely, decyltrimethylammonium bromide [N10111][Br] and 1-decyl-3-methylimidazolium chloride [C10mim][Cl] as cationic surfactant ionic liquids and cholinium decanoate [Chl][Dec], an anionic surfactant ionic liquid, on the structure and function of laccase from the fungus Trametes versicolor (TvL) by the molecular dynamics (MD) simulation method. In summary, this study showed that laccase solvent-accessible surface area increased in the ionic liquid [Chl][Dec] while it decreased in the other two ionic liquids. Interestingly, [Chl][Dec] ionic liquid components formed hydrogen bonds with laccase, while [N10111][Br] and [C10mim][Cl] components were unable to form hydrogen bonds with laccase. The quantity of hydrogen bonds formed between water molecules and the enzyme was also diminished in the presence of [Chl][Dec] in comparison to the other two ionic liquids. especially at a concentration of 250 mM. In 250 mM concentrations of [N10111][Br] and [C10mim][Cl], clusters of long-chain cations are likely to form near the copper T1 site. However, even at low [Chl][Dec] concentrations, long [Dec]- chains were observed to penetrate the enzyme near the copper T1 site, and at 250 mM [Chl][Dec], a large cluster of anions occupied the opening of the active site. The results of the analysis also show that the interaction between the [Dec]- anion and the enzyme is stronger than the interaction between [N10111]+ and [C10mim]+ with laccase; in addition, the [Dec]- anion, compared to [Br]- and [Cl]- has a much greater tendency to bind with the enzyme residues.

Mechanism of non-phenolic substrate oxidation by the fungal laccase Type 1 copper site from Trametes versicolor: the case of benzo[a]pyrene and anthracene.

Laccases (EC 1.10.3.2) are multicopper oxidases with the capability to oxidize diverse phenolic and non-phenolic substrates. While the molecular mechanism of their activity towards phenolic substrates is well-established, their reactivity towards non-phenolic substrates, such as polycyclic aromatic hydrocarbons (PAHs), remains unclear. To elucidate the oxidation mechanism of PAHs, particularly the activation mechanism of the sp2 aromatic C-H bond, we conducted a density functional theory investigation on the oxidation of two PAHs (anthracene and benzo[a]pyrene) using an extensive model of the T1 copper catalytic site of the fungal laccase from Trametes versicolor.

Ligninolytic enzyme potential of Trametes spp. associated with leaf litter in riparian forest of the Amazônia region.

The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.

Synthesis, characterization, and phosphorus adsorption of Mg/Fe-modified biochar from cotton stalk pretreated with Coriolus versicolor.

In recent years, the research potential in utilizing biochars as adsorbents in adsorption processes has grown due to their eco-friendly and economical nature. However, biochar often possesses a negative surface charge that limits its affinity for binding anions. Nitric acid washing and pretreatment with Coriolus versicolor can break down the lignocellulosic structure in cotton stalk waste, facilitating the subsequent impregnation of Mg and Fe metal oxides. These pretreatment steps can lead to the production of diverse and functionalized biochars with higher adsorption capacities. In this study, cotton stalk waste was first washed with diluted nitric acid and then subjected to biological pretreatment by incubation with C. versicolor, followed by impregnation with Mg and Fe to obtain CV-CS/Fe and CV-CS/Mg biochars. The results showed that the applied pretreatments altered the physicochemical properties and significantly increased the phosphorus adsorption capacity. The adsorption capacities of CV-CS/Fe and CV-CS/Mg biochars were found to be 277.88 and 507.01 mg g-1, respectively. The results indicate that the incorporation of multiple metal oxide impregnates enhances P adsorption. Furthermore, in the kinetic study, pseudo-first-order and pseudo-second-order models provided a well fit, determining chemical adsorption as the main adsorption mechanism for phosphorus adsorption. The biochars demonstrated compatibility with Langmuir-Freundlich models. Overall, the findings suggest the possibility of synthesizing biochars with improved adsorptive properties through pretreatment, and these engineered biochars hold promising potential as effective adsorbents in the field. PRACTITIONER POINTS: Eco-friendly, natural, and economical biochar was synthesized. Biochar was produced via Coriolus versicolor pretreatment. High adsorption capacities of CV-PS/Mg biochars were found to be 507.01 mg g-1. Adsorption capacities of biochars can be improved by pretreatment.

Enhanced biodegradation of benzo[a]pyrene with Trametes versicolor stimulated by citric acid.

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants with carcinogenic, mutagenic and teratogenic effects. The white-rot fungi in the fungal group have significant degradation ability for high molecular weight organic pollutants. However, exogenous fungi are easily antagonized by indigenous microorganisms. Low molecular weight organic acids, a small molecular organic matter secreted by plants, can provide carbon sources for soil microorganisms. Combining organic acids with white rot fungi may improve the nutritional environment of fungi. In this study, immobilized Trametes versicolor was used to degrade benzo[a]pyrene in soil, and its effect on removing benzo[a]pyrene in soil mediated by different low molecular weight organic acids was investigated. The results showed that when the degradation was 35 days, the removal effect of the experimental group with citric acid was the best, reaching 43.7%. The degradation effect of Trametes versicolor on benzo[a]pyrene was further investigated in the liquid medium when citric acid was added, and the effects of citric acid on the biomass, extracellular protein concentration and laccase activity of Trametes versicolor were investigated by controlling different concentrations of citric acid. In general, citric acid can act as a carbon source for Trametes versicolor and promote its extracellular protein secretion and laccase activity, thereby accelerating the mineralization of benzo[a]pyrene by Trametes versicolor. Therefore, citric acid can be used as a biostimulant in the remediation of PAHs contaminated soil with Trametes versicolor.

The role and molecular mechanism of Trametes Robiniophila Murr(Huaier) in tumor therapy.

ETHNOPHARMACOLOGICAL RELEVANCE: Trametes Robiniophila Murr, commonly known as Huaier, has been extensively documented in ethnopharmacology research in China. Huaier has a long history of clinical usage spanning over 1000 years in China. Traditional clinical application records demonstrate the wide utilization of Huaier for treating various cancers and enhancing the autoimmunity of tumor patients. AIM OF THE REVIEW: The present study provides a comprehensive review of the traditional uses, phytochemical constituents, pharmacological activities, anti-tumor mechanism, and potential applications of Huaier, thereby offering valuable insights for the further development and utilization of this natural product. MATERIALS AND METHODS: This study employed the keywords "Trametes Robiniophila Murr" and "Huaier" to retrieve relevant information on Huaier from various databases, including PubMed, Web of Science, Springer, Science Direct, ACS, Wiley, CNKI, Baidu Scholar, Google Scholar, and ancient materia medica. RESULTS: Trametes Robiniophila Murr (Huaier), a traditional Chinese medicine, has demonstrated significant efficacy in the clinical treatment of various tumors. The primary bioactive constituents of Huaier consist of fungal-derived compounds, including polysaccharides, proteins, ketones, alkaloids, and minerals. The research findings demonstrate that Huaier serves as a reliable adjunctive therapeutic agent by effectively inhibiting cancer cell proliferation, inducing apoptosis in cancer cells, suppressing tumor metastasis, regulating tumor stem cells and immune function. Therefore, it exerts a potent anti-tumor effect when used in conjunction with conventional anti-cancer therapies. CONCLUSIONS: The analysis of traditional uses, phytochemical composition, and pharmacological activity reveals that Huaier exhibits significant potential as a medicinal plant with diverse pharmacological effects. Owing to its numerous advantages, Huaier holds immense promise for application in the domains of tumor prevention and treatment, enhancing both survival time and quality of life among cancer patients.

Application of fungal-based microbial fuel cells for biodegradation of pharmaceuticals: Comparative study of individual vs. mixed contaminant solutions.

The present study focuses on the application of fungal-based microbial fuel cells (FMFC) for the degradation of organic pollutants including Acetaminophen (APAP), Para-aminophenol (PAP), Sulfanilamide (SFA), and finally Methylene Blue (MB). The objective is to investigate the patterns of degradation (both individually and as a mixture solution) of the four compounds in response to fungal metabolic processes, with an emphasis on evaluating the possibility of generating energy. Linear Sweep Voltammetry (LSV) has been used for electrochemical analysis of the targeted compounds on a Glassy Carbon Electrode (GCE). A dual chamber MFC has been applied wherein the cathodic compartment, the reduction reaction of oxygen was catalyzed by an elaborated biofilm of Trametes trogii, and the anodic chamber consists of a mixed solution of 200 mg L-1 APAP, PAP, MB, and SFA in 0.1 M PBS and an elaborated biofilm of Trichoderma harzianum. The obtained results showed that all the tested molecules were degraded over time by the Trichoderma harzianum. The biodegradation kinetics of all the tested molecules were found to be in the pseudo-first-order. The results of half-lives and the degradation rate reveal that APAP in its individual form degrades relatively slower (0.0213 h-1) and has a half-life of 33 h compared to its degradation in a mixed solution with a half-life of 20 h. SFA showed the longest half-life in the mixed condition (98 h) which is the opposite of its degradation as individual molecules (20 h) as the fastest molecule compared to other pollutants. The maximum power density of the developed MFC dropped from 0.65 mW m-2 to 0.32 mW m-2 after 45.5 h, showing that the decrease of the residual concentration of molecules in the anodic compartment leads to the decrease of the MFC performance.

Two Zn2Cys6-type transcription factors respond to aromatic compounds and regulate the expression of laccases in the white-rot fungus Trametes hirsuta.

White-rot fungi differentially express laccases when they encounter aromatic compounds. However, the underlying mechanisms are still being explored. Here, proteomics analysis revealed that in addition to increased laccase activity, proteins involved in sphingolipid metabolism and toluene degradation as well as some cytochrome P450s (CYP450s) were differentially expressed and significantly enriched during 48 h of o-toluidine exposure, in Trametes hirsuta AH28-2. Two Zn2Cys6-type transcription factors (TFs), TH8421 and TH4300, were upregulated. Bioinformatics docking and isothermal titration calorimetry assays showed that each of them could bind directly to o-toluidine and another aromatic monomer, guaiacol. Binding to aromatic compounds promoted the formation of TH8421/TH4300 heterodimers. TH8421 and TH4300 silencing in T. hirsuta AH28-2 led to decreased transcriptional levels and activities of LacA and LacB upon o-toluidine and guaiacol exposure. EMSA and ChIP-qPCR analysis further showed that TH8421 and TH4300 bound directly with the promoter regions of lacA and lacB containing CGG or CCG motifs. Furthermore, the two TFs were involved in direct and positive regulation of the transcription of some CYP450s. Together, TH8421 and TH4300, two key regulators found in T. hirsuta AH28-2, function as heterodimers to simultaneously trigger the expression of downstream laccases and intracellular enzymes. Monomeric aromatic compounds act as ligands to promote heterodimer formation and enhance the transcriptional activities of the two TFs.IMPORTANCEWhite-rot fungi differentially express laccase isoenzymes when exposed to aromatic compounds. Clarification of the molecular mechanisms underlying differential laccase expression is essential to elucidate how white-rot fungi respond to the environment. Our study shows that two Zn2Cys6-type transcription factors form heterodimers, interact with the promoters of laccase genes, and positively regulate laccase transcription in Trametes hirsuta AH28-2. Aromatic monomer addition induces faster heterodimer formation and rate of activity. These findings not only identify two new transcription factors involved in fungal laccase transcription but also deepen our understanding of the mechanisms underlying the response to aromatics exposure in white-rot fungi.

Biofoams with untapped enzymatic potential produced from beer bagasse by indigenous fungal strains.

White rot fungi are promising organisms for the production of mycelial-based biofoams, providing a sustainable means of valorizing lignocellulosic wastes. This study explores the utilization of two indigenous fungal species, isolated from Argentina and belonging to the genera Trametes, for producing biofoams from brewery waste. The resulting biofoams exhibited an average density of 0.30 g cm-3, a Young's modulus of approximately 1 MPa, and a compressive stress of around 19 MPa. Additionally, the variation of laccase activity throughout the biofoam production process was evaluated. Surprisingly, residual laccase activity was detected in the biofoams following oven drying at temperatures of 60, 80, and 100 °C. This detection highlights the untapped enzymatic potential of the biofoams and positions them as promising green catalysts for various biotechnological applications.

Improved productivity and dye removal performance of Trametes versicolor pellets using rice husk as a co-substrate.

Pellet production represents a critical step for several processes requiring fungal biomass, nevertheless, its optimization is seldom reported. The use of finely ground rice husk as a microcarrier and co-substrate permitted a marked increase (≈ 2.7×) in the productivity of fungal pellet production using Trametes versicolor compared to traditional production methods. The pellets show similar structure and smaller size compared to typical sole-mycelium pellets, as well as comparable laccase activity. The efficiency of the pellets for biodegradation was confirmed by the removal of the crystal violet dye, achieving significantly faster decolorization rates compared to the traditionally produced pellets. The use of these pellets during the continuous treatment of the dye in a stirred tank bioreactor resulted in 97% decolorization operating at a hydraulic residence time of 4.5 d.

Development of binderless fiberboard from poplar wood residue with Trametes hirsuta.

The utilization of agricultural and forestry residues for the development and preparation of green binderless fiberboard (BF) is an effective way to realize high-value utilization of lignocellulose biomass resources. This study focuses on the fabrication of BF with excellent mechanical and waterproof properties, utilizing poplar wood residue (PWR) as raw material and Trametes hirsuta as a pretreatment method. During the fermentation process, lignin-degrading enzymes and biological factors, such as sugars, were produced by T. hirsuta, which activated lignin by depolymerizing lignin bonds and modifying structural functional groups, and forming new covalent bonds between poplar fibers, ultimately enhancing adhesion. Additionally, the activated lignin molecules and sugar molecules coalesce under high temperatures and pressures, forming a dense carbonization layer that bolsters the mechanical properties of the fiberboard and effectively shields it from rapid water infiltration. The bio-pretreated BF for 10 days shows a MOR and MOE of up to 36.1 Mpa and 3704.3 Mpa, respectively, which is 261% and 247.8% higher than that of the bio-untreated fiberboard, and the water swelling ratio (WSR) rate is only 5.6%. Chemical composition analysis revealed that repolymerization occurred among lignin, cellulose, and hemicellulose, especially the molecular weight of lignin changed significantly, with the Mw of lignin increasing from 312066 g/mol to 892362 g/mol, and then decreasing to 825021 g/mol. Mn increased from 277790 g/mol to 316987.5 g/mol and then decreased to 283299.5 g/mol at 21 days. Compared to other artificial fiberboards prepared through microbial pretreatment, the BF prepared by microorganisms in this study exhibited the highest mechanical properties among the poplar wood biobased panels.

Evaluation of the Hepatoprotective Effect of the Lumpy Bracket Medicinal Mushroom Trametes gibbosa (Agaricomycetes) on CCl4-Induced Liver Injury in Rats.

Mushrooms have been used as medicine by humans for more than 5000 years. They have had a successful role in treating immune deficiencies. Nowadays, some extracts and compounds obtained from medicinal mushrooms have increased a great prospect of treating many disorders by having a great role in modulation of immune system, cancer inhibiting, cardio-vascular health, antiviral, antibacterial, antioxidant and protective effects against hepatitis and diabetes. In this study, we evaluated the antioxidant effect of methanol and hot water extract of the Trametes gibbosa (Pers.) Fr. mushroom and hepatoprotective effect of the extract with the most radical scavenging potency. To assess the antioxidant properties of different extracts of the mushroom, DPPH method was used. For assessing the hepatoprotective properties, a seven-day experiment was designed, and liver toxicity was induced by carbon tetrachloride [intraperitoneal (ip) for 7 consecutive days, 0.5 mL/kg body weight (BW)]. Rats were simultaneously fed with aqueous extract of the mushroom with the dose of 250, 500, and 1000 mg/kg BW and silymarin (100 mg/kg BW) as positive control. At the end of the experiment, blood serums of the rats were collected for quantification of major liver factors (e.g., aspartate aminotransferase, alanine aminotransferase, alanine phosphatase, bilirubin, etc.). Tissue samples were obtained for pathological examination. Based on the results, the aqueous extract showed more potent radical scavenging activity (half-maximal inhibitory concentration = 414.33 µg/mL, compared with 936.92 µg/mL for methanolic extract). Indeed, hepatoprotective properties of the aqueous extract of the mushroom (500 and 1000 mg/kg BW) were comparable with those of silymarin and even showed superior protective effects in histopathological examination. It seems that with further complementary studies, T. gibbosa could be considered a potential candidate for hepatoprotection.

Assessment of textile effluent treatment by immobilized Trametes pubescens MB 89 for plant growth promotion.

Industrialization and the ever-increasing world population have diminished high-quality water resources for sustainable agriculture. It is imperative to effectively treat industrial effluent to render the treated water available for crop cultivation. This study aimed to assess the effectiveness of textile effluent treated with Trametes pubescens MB 89 in supporting maize cultivation. The fungal treatment reduced the amounts of Co, Pb and As in the textile effluent. The biological oxygen demand, total dissolved solids and total suspended solids were within the permissible limits in the treated effluent. The data indicated that the irrigation of maize with fungal-treated textile effluent improved the growth parameters of the plant including root, shoot length, leaf area and chlorophyll content. Moreover, better antioxidant activity, total phenol content and protein content in roots, stems and leaves of maize plants were obtained. Photosynthetic parameters (potential quantum yield, electron transport rate and fluorescence yield of non-photochemical losses other than heat) were also improved in the plants irrigated with treated effluent as compared to the control groups. In conclusion, the treatment of textile effluent with the immobilized T. pubescens presents a sustainable solution to minimize chemical pollution and effectively utilize water resources.

Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

BACKGROUND: Gemcitabine is the first-line chemotherapy drug that can easily cause chemotherapy resistance. Huaier is a traditional Chinese medicine and shows an antitumor effect in pancreatic cancer, but whether it can enhance the gemcitabine chemotherapeutic response and the potential mechanism remain unknown. PURPOSE: This study was performed to explore the effect of Huaier in promoting the tumor-killing effect of gemcitabine and elucidate the possible mechanism in pancreatic cancer. METHODS: Cell Counting Kit-8 assays and colony formation assays were used to detect proliferation after different treatments. Protein coimmunoprecipitation was applied to demonstrate protein interactions. Nuclear protein extraction and immunofluorescence were used to confirm the intracellular localization of the proteins. Western blotting was performed to detect cell proliferation-related protein expression or cancer stem cell-associated protein expression. Sphere formation assays and flow cytometry were used to assess the stemness of pancreatic cancer cells. The in vivo xenograft model was used to confirm the inhibitory effect under physiological conditions, and immunohistochemistry was used to detect protein expression. RESULTS: Huaier suppressed the proliferation and stem cell-like properties of pancreatic cancer cells. We found that Huaier suppressed the expression of forkhead box protein M1 (FoxM1). In addition, Huaier inhibited FoxM1 function by blocking its nuclear translocation. Treatment with Huaier reversed the stemness induced by gemcitabine in a FoxM1-dependent manner. Furthermore, we verified the above results by an in vivo study, which reached the same conclusion as those in vitro. CONCLUSION: Overall, this study illustrates that Huaier augments the tumor-killing effect of gemcitabine through suppressing the stemness induced by gemcitabine in a FoxM1-dependent way. These results indicate that Huaier can be applied to overcome gemcitabine resistance.

Morphological, sterical, and localized thermodynamics in the adsorption of CO2 by activated biocarbon from the white rot fungi Trametes gibbosa.

The capture of CO2 by biochar has recently become one of the cornerstones of circular economy models for a sustainable society. In this work, we synthesized an activated biocarbon using Trametes gibbosa (BioACTG) in a one-step synthesis. We investigated CO2 adsorption mechanisms under five different temperatures using a statistical physics approach. The data was better represented by the multilayer model with two distinguished energies, providing more accurate values for the estimated parameters. According to the number of carbon dioxide molecules per site (n) and the densities of the receptor sites (Dzif), the tendency to form a second layer increased as the temperature increased. The adsorption of CO2 on BioACTG was exothermic (the values of Qasat = 15.5 mmol/g at 273 K decrease to 10.5 mmol/g at 353 K), and the temperature influenced CO2 as well as the morphological features of the process. A computational approach was used to investigate the electronic properties of the adsorbate, showing that its lowest unoccupied orbital (LUMO) heavily contributed to the high efficiency of the process which was ruled by pore diffusion mechanisms driven by energetic fluctuations. Other molecules present in CO2-rich mixtures were also investigated, showing that their concentration limited their competitiveness with CO2.

Using low-shear aerated and agitated bioreactor for producing two specific laccases by trametes versicolor cultures induced by 2,5-xylidine: Process development and economic analysis.

Laccase isoforms from basidiomycetes exhibit a superior redox potential compared to commercially available laccases obtained from ascomycete fungi, rendering them more reactive toward mono-substituted phenols and polyphenolic compounds. However, basidiomycetes present limitations for large-scale culture in liquid media, restraining the current availability of laccases from this fungal class. To advance laccase production from basidiomycetes, a newly designed 14-L low-shear aerated and agitated bioreactor provided enzyme titers up to 23.5 IU/mL from Trametes versicolor cultures. Produced enzymes underwent ultrafiltration and LC/MS-MS characterization, revealing the predominant production of only two out of the ten laccases predicted in the T. versicolor genome. Process simulation and economic analysis using SuperPro designer® suggested that T. versicolor laccase could be produced at US$ 3.60/kIU in a 200-L/batch enterprise with attractive economic parameters and a payback period of 1.7 years. The study indicates that new bioreactors with plain design help to produce low-cost enzymes from basidiomycetes.

Structure elucidation and antiviral activity of a cold water-extracted mannogalactofucan Ts1-1A from Trametes sanguinea against human cytomegalovirus in vitro.

In this study, we purified a partially acetylated heteropolysaccharide (Ts1-1A) from the fruit bodies of Trametes sanguinea Lloyd through cold water extraction and serial chromatographic separation. The purified polysaccharide Ts1-1A (12.8 kDa) was characterized as a branched mannogalactofucan with a backbone of alternately connected 1,3-linked α-Fucp and 1,6-linked α-Galp, which was partially substituted by non-reducing end units of ß-Manp at O-2 and O-3 positions of 1,6-linked α-Galp. Ts1-1A showed pronounced anti-human cytomegalovirus activity at the concentration of 200 and 500 µg/mL in systematical assessments including morphological changes, western blotting, qPCR, indirect immunofluorescence and tissue culture infective dose assays. Moreover, Ts1-1A exerted its antiviral activity at two distinct stages of viral proliferation manifesting as significantly inhibiting viral protein (IE1/2 and p52) expression and reducing viral gene (UL123, UL44 and UL32) replication in the HCMV-infected WI-38 cells. At viral attachment stage, Ts1-1A interacted with HCMV and prevented HCMV from attaching to its host cells. While at early phase of viral replication stage, Ts1-1A suppressed HCMV replication by downregulating NQO1 and HO-1 proteins related to oxidative stress as an antioxidant. To sum up, Ts1-1A is a promising anti-HCMV agent which could be developed for HCMV infection prevention and therapy.

Comparative Analysis of Viromes Identified in Multiple Macrofungi.

Macrofungi play important roles in the soil elemental cycle of terrestrial ecosystems. Fungal viruses are common in filamentous fungi, and some of them can affect the growth and development of hosts. However, the composition and evolution of macrofungal viruses are understudied. In this study, ninety strains of Trametes versicolor, Coprinellus micaceus, Amanita strobiliformis, and Trametes hirsuta were collected in China. Four mixed pools were generated by combining equal quantities of total RNA from each strain, according to the fungal species, and then subjected to RNA sequencing. The sequences were assembled, annotated, and then used for phylogenetic analysis. Twenty novel viruses or viral fragments were characterized from the four species of macrofungi. Based on the phylogenetic analysis, most of the viral contigs were classified into ten viral families or orders: Barnaviridae, Benyviridae, Botourmiaviridae, Deltaflexiviridae, Fusariviridae, Hypoviridae, Totiviridae, Mitoviridae, Mymonaviridae, and Bunyavirales. Of these, ambi-like viruses with circular genomes were widely distributed among the studied species. Furthermore, the number and overall abundance of viruses in these four species of macrofungi (Basidiomycota) were found to be much lower than those in broad-host phytopathogenic fungi (Ascomycota: Sclerotinia sclerotiorum, and Botrytis cinerea). By employing metatranscriptomic analysis in this study, for the first time, we demonstrated the presence of multiple mycoviruses in Amanita strobiliformis, Coprinellus micaceus, Trametes hirsute, and Trametes versicolor, significantly contributing to research on mycoviruses in macrofungi.