RESUMEN
BACKGROUND: Solid-state fermentation is a fungal culture technique used to produce compounds and products of industrial interest. The growth behaviour of filamentous fungi on solid media is challenging to study due to the intermixity of the substrate and the growing organism. Several strategies are available to measure indirectly the fungal biomass during the fermentation such as following the biochemical production of mycelium-specific components or microscopic observation. The microscopic observation of the development of the mycelium, on lignocellulosic substrate, has not been reported. In this study, we set up an experimental protocol based on microscopy and image processing through which we investigated the growth pattern of Phanerochaete chrysosporium on different Miscanthus x giganteus biomass fractions. RESULTS: Object coalescence, the occupied surface area, and radial expansion of the colony were measured in time. The substrate was sterilized by autoclaving, which could be considered a type of pre-treatment. The fastest growth rate was measured on the unfractionated biomass, followed by the soluble fraction of the biomass, then the residual solid fractions. The growth rate on the different fractions of the substrate was additive, suggesting that both the solid and soluble fractions were used by the fungus. Based on the FTIR analysis, there were differences in composition between the solid and soluble fractions of the substrate, but the main components for growth were always present. We propose using this novel method for measuring the very initial fungal growth by following the variation of the number of objects over time. Once growth is established, the growth can be followed by measurement of the occupied surface by the mycelium. CONCLUSION: Our data showed that the growth was affected from the very beginning by the nature of the substrate. The most extensive colonization of the surface was observed with the unfractionated substrate containing both soluble and solid components. The methodology was practical and may be applied to investigate the growth of other fungi, including the influence of environmental parameters on the fungal growth.
Asunto(s)
Phanerochaete/crecimiento & desarrollo , Biomasa , Fermentación , Cinética , Micelio/crecimiento & desarrollo , Micelio/metabolismo , Phanerochaete/química , Phanerochaete/metabolismo , Poaceae/crecimiento & desarrollo , Poaceae/metabolismoRESUMEN
Microorganisms capable of decomposing cellulose, xylan, starch and protein were individually isolated from swine manure compost and soil in this study. The correlations with pH, carbon source concentration, C/N ratio and enzyme activity among these isolated microorganisms were also investigated. Furthermore, the effect of additional inoculation in the compost was studied by measuring variations in the C/N ratio, enzyme activity and compost maturation rate. The inoculated microorganisms used in this study included four bacterial isolates and one commercial microorganism Phanerochaete chrysosporium. The results indicated that the isolated Kitasatospora phosalacinea strain C1, which is a cellulose-degraded microorganism, presented the highest enzyme activity at 31 â and pH 5.5, while the C/N ratio was 0.8%. The isolated xylan-degraded microorganism Paenibacillus glycanilyticus X1 had the highest enzyme activity at 45 â and pH 7.5, while the C/N ratio was 0.5%. The starch-degraded microorganism was identified as Bacillus licheniformis S3, and its highest enzyme activities were estimated to be 31 â and pH 7.5 while the C/N ratio was 0.8%. The highest enzyme activity of the protein-degraded microorganism Brevinacillus agri E4 was obtained at 45 â and pH 8.5, while the C/N ratio was 1.0%. The rate of temperature increase in the compost inoculated with P. chrysosporium was only higher than that of the compost without inoculation, and its compost maturation level was also lower than that of other composts with additional inoculation. The optimal initial C/N ratio of the compost was 27.5 and the final C/N ratio was 18.9. The composting results also indicated that the secondary inoculation would benefit compost maturation, and the lowest final C/N ratio of 17.0 was obtained.
Asunto(s)
Bacterias/crecimiento & desarrollo , Compostaje , Estiércol/microbiología , Oryza/microbiología , Phanerochaete/crecimiento & desarrollo , Animales , PorcinosRESUMEN
Cr(VI) is one of the most common environmental pollutants. The non-biodegradable Cr(VI) in aqueous solution accumulates along the food chain and damages the health of plant, animal, and human. In this study, solid-state fermentation technology was used to treat Caulis lonicerae residue to improve its adsorption capacity for Cr(VI). Caulis lonicerae can be used to extract the active ingredients such as organic acids, flavonoids, and triterpenoid saponins that have various effects including anti-oxidation and immune boosting. However, there is no proper treatment for large amount of residue left after extraction. The Phanerochaete chrysosporium was inoculated into the residue for solid-state fermentation, and the adsorption capacity of C. lonicerae residue before (CLr) and after fermentation (FCLr) regarding Cr(VI) adsorption was compared. In the range of 40-120 mg/L Cr(VI), the adsorption capacity of FCLr was significantly higher than that of CLr. The scanning electron microscopy (SEM) results exhibited a rougher surface of FCLr. The Fourier-transform infrared spectroscopy (FTIR) results revealed that the structure of FCLr was affected by the combination of Cr(VI), and the multiple functional groups interacted with Cr(VI) (such as -OH, C-H, C = O, C-O-C, and C-O). The adsorption capacity could reach 48.91 mg/g and the Cr(VI) removal percentage may reach 98.07% for FCLr increased by 28.24% through fermentation.
Asunto(s)
Cromo/metabolismo , Lonicera/química , Phanerochaete/crecimiento & desarrollo , Contaminantes Químicos del Agua/metabolismoRESUMEN
To assess the relationship between resource use and hyphal growth in a cord-forming basidiomycete, Phanerochaete velutina, soil microcosm experiments were conducted using wood blocks of three different sizes in three different soil quantities, thereby simulating the different amounts of available nutrients. The highest percentage weight loss was observed in the smallest wood blocks after a 27-d incubation period in soil microcosms, although the percentage weight loss over the 2-month pure culture colonization prior to inoculation was not significantly different among various block sizes. The greatest hyphal outgrowth was also observed in the smallest wood blocks and was positively associated with wood decay. The slopes of the regression lines between hyphal coverage and percentage wood mass loss were identical among different wood sizes, but the slopes between hyphal coverage and absolute wood mass loss were steeper in the smaller wood blocks than that in largest one. These results suggest that the level of intensity of mycelial foraging for new resources in the soil depends on the percentage of the amount of wood resource utilized, and not on the absolute amount of carbon obtained from the wood.
Asunto(s)
Hifa/crecimiento & desarrollo , Phanerochaete/crecimiento & desarrollo , MaderaRESUMEN
Skin darkening results as a consequence of the accumulation of skin pigment melanin. To combat this, the amplitude of skin lightening agents are commercially available, most of which inhibit melanin synthesis. Decolorization of melanin is an alternative method of skin lightening. In this study, we show that lignin peroxidase (LiP), an extracellular enzyme purified from Phanerochaete chrysosporium NK-1 isolated from a forest soil can effectively degrade and decolorize melanin in vitro. Decolorization conditions including pH, temperature, incubation time, enzyme concentration, and mediator addition were investigated to optimize the reaction conditions. The results indicate that pH 3, 40 °C, 15 IU/ml, and 10 h incubation were the optimal conditions for the decolorization of the melanin. The use of the mediator, veratryl alcohol was also found effective to enhance the efficacy of the melanin decolonization, with up to 92% decolorization. The scanning electron microscopy results showed void spaces on the treated melanin granules as compared to the untreated sample, indicating the degradation of melanin. Changes in the fingerprint region of the melanin were observed. Between wavenumbers 1500-500 cm-1, for example, the presence of new peaks in the treated melanin at 1513, 1464, and 1139 cm-1 CH2, CH3 bend and C-O-C stretch represented structural changes. A new peak at 2144 cm-1 (alkynyl C≡C stretch) was also detected in the decolorized melanin. The cytotoxicity study has shown that the treated melanin and LiP have low cytotoxic effects; however, the mediator of veratryl alcohol could result in high mortality which suggests that its use should be meticulously tested in formulating health and skincare products. The findings of the study suggest that LiP produced by Phanerochaete chrysosporium has the potential to be used in the medical and cosmetic industries, particularly for the development of biobased cosmetic whitening agents.
Asunto(s)
Melaninas/química , Peroxidasas/farmacología , Phanerochaete/aislamiento & purificación , Preparaciones para Aclaramiento de la Piel/farmacología , Animales , Artemia/efectos de los fármacos , Artemia/crecimiento & desarrollo , Alcoholes Bencílicos/química , Alcoholes Bencílicos/toxicidad , Cosméticos , Bosques , Proteínas Fúngicas/farmacología , Proteínas Fúngicas/toxicidad , Humanos , Concentración de Iones de Hidrógeno , Microscopía Electrónica de Rastreo , Peroxidasas/toxicidad , Phanerochaete/enzimología , Phanerochaete/crecimiento & desarrollo , Proteolisis , Preparaciones para Aclaramiento de la Piel/toxicidad , Microbiología del Suelo , Factores de TiempoRESUMEN
The Target Of Rapamycin (TOR) signaling pathway is known to regulate growth in response to nutrient availability and stress in eukaryotic cells. In the present study, we have investigated the TOR pathway in the white-rot fungus Phanerochaete chrysosporium. Inhibition of TOR activity by rapamycin affects conidia germination and hyphal growth highlighting the conserved mechanism of susceptibility to rapamycin. Interestingly, the secreted protein content is also affected by the rapamycin treatment. Finally, homologs of the components of TOR pathway can be identified in P. chrysosporium. Altogether, those results indicate that the TOR pathway of P. chrysosporium plays a central role in this fungus.
Asunto(s)
Proteínas Fúngicas/metabolismo , Phanerochaete/crecimiento & desarrollo , Phanerochaete/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Sitios de Unión , Proteínas Fúngicas/antagonistas & inhibidores , Humanos , Enlace de Hidrógeno , Reacción en Cadena de la Polimerasa , Estructura Secundaria de Proteína , Proteoma , Transducción de Señal/efectos de los fármacos , Sirolimus/farmacología , Esporas Fúngicas/metabolismo , Serina-Treonina Quinasas TOR/antagonistas & inhibidores , beta-Glucosidasa/metabolismoRESUMEN
The root-knot nematode Meloidogyne incognita has a wide host range and it is one of the most economically important crop parasites worldwide. Biological control has been a good approach for reducing M. incognita infection, for which many nematophagous fungi are reportedly applicable. However, the controlling effects of Phanerochaete chrysosporium strain B-22 are still unclear. In the present study we characterized the parasitism of this strain on M. incognita eggs, second-stage juveniles (J2), and adult females. The highest corrected mortality was 71.9% at 3 × 108 colony forming units (CFU) mL-1 and the estimated median lethal concentration of the fungus was 0.96 × 108 CFU mL-1. Two days after treatment with Phanerochaete chrysosporium strain B-22 eggshells were dissolved. A strong lethal effect was noted against J2, as the fungal spores developed in their body walls, germinated, and the resulting hyphae crossed the juvenile cuticle to dissolve it, thereby causing shrinkage and deformation of the juvenile body wall. The spores and hyphae also attacked adult females, causing the shrinkage and dissolution of their bodies and leakage of contents after five days. Greenhouse experiments revealed that different concentrations of the fungal spores effectively controlled M. incognita. In the roots, the highest inhibition rate for adult females, juveniles, egg mass, and gall index was 84.61%, 78.91%, 84.25%, and 79.48%, respectively. The highest juvenile inhibition rate was 89.18% in the soil. Phanerochaete chrysosporium strain B-22 also improved tomato plant growth, therefore being safe for tomato plants while effectively parasitizing M. incognita. This strain is thus a promising biocontrol agent against M. incognita.
Asunto(s)
Phanerochaete/crecimiento & desarrollo , Enfermedades de las Plantas/microbiología , Tylenchoidea/microbiología , Animales , Antinematodos/farmacología , Femenino , Interacciones Huésped-Parásitos/efectos de los fármacos , Hifa/efectos de los fármacos , Hifa/crecimiento & desarrollo , Hifa/patogenicidad , Phanerochaete/efectos de los fármacos , Phanerochaete/patogenicidad , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiologíaRESUMEN
Endophytic fungi live inside vegetal tissues without causing damage to the host plant and may provide lead compounds for drug discovery. The co-culture of two or more endophytic fungi can trigger silent gene clusters, which could lead to the isolation of bioactive compounds. In this study, two endophytic strains isolated from Handroanthus impetiginosus leaves, identified as Talaromyces purpurogenus H4 and Phanerochaete sp. H2, were grown in mixed and axenic cultures. The meroterpenoid austin was detected only in the extracts from the mixed culture. Once isolated, austin displayed very interesting trypanocidal activity, with an IC50 value of 36.6 ± 1.2 µg/mL against Trypanosoma cruzi in the epimastigote form. The results obtained highlight the importance of the co-culturing of endophytic fungi to obtain natural bioactive products. The findings also enhance our understanding of the ecological relationships between endophytic fungi.
Asunto(s)
Endófitos/crecimiento & desarrollo , Tabebuia/microbiología , Talaromyces/crecimiento & desarrollo , Talaromyces/metabolismo , Tripanocidas/metabolismo , Técnicas de Cocultivo , Endófitos/química , Endófitos/genética , Phanerochaete/química , Phanerochaete/genética , Phanerochaete/crecimiento & desarrollo , Phanerochaete/metabolismo , Hojas de la Planta/microbiología , Talaromyces/química , Talaromyces/genética , Terpenos/análisis , Terpenos/metabolismo , Terpenos/farmacología , Tripanocidas/análisis , Tripanocidas/farmacología , Trypanosoma cruzi/efectos de los fármacos , Trypanosoma cruzi/crecimiento & desarrolloRESUMEN
The simultaneous removal of phenol and selenite from synthetic wastewater was investigated by adopting two different co-culturing techniques using the fungus Phanerochaete chrysosporium and the bacterium Delftia lacustris. Separately grown biomass of the fungus and the bacterium (suspended co-culture) was incubated with different concentrations of phenol (0-1,200 mg/L) and selenite (10 mg/L). The selenite ions were biologically reduced to extracellular Se(0) nanoparticles (3.58 nm diameter) with the simultaneous degradation of up to 800 mg/L of phenol. Upon growing the fungus and the bacterium together using an attached growth co-culture, the bacterium grew as a biofilm onto the fungus. The extracellularly produced Se(0) in the attached growth co-culture had a minimum diameter of 58.5 nm. This co-culture was able to degrade completely 50 mg/L phenol, but was completely inhibited at a phenol concentration of 200 mg/L.
Asunto(s)
Delftia/metabolismo , Phanerochaete/metabolismo , Fenol/metabolismo , Ácido Selenioso/metabolismo , Selenio/metabolismo , Biodegradación Ambiental , Biotransformación , Técnicas de Cocultivo , Delftia/crecimiento & desarrollo , Oxidación-Reducción , Phanerochaete/crecimiento & desarrollo , Aguas Residuales/microbiología , Contaminantes del Agua/metabolismoRESUMEN
As a persistent organic pollutant listed in the Stockholm Convention, perfluorooctane sulfonate (PFOS) is extremely refractory to degradation under ambient conditions. Its potential ecotoxicity has aroused great concerns and research interests. However, little is known about the toxicity of PFOS on fungus. In this study, the white rot fungus Phanerochaete chrysosporium (P. chrysosporium) was adopted to assess the toxicity of PFOS in liquid culture. The addition of 100â¯mg/L PFOS potassium salt significantly decreased the fungal biomass by up to 76.4% comparing with un-amended control during the incubation period. The hyphostroma of P. chrysosporium was wizened and its cell membrane was thickened, while its vesicle structure was increased, based on the observation with scanning electron microscope (SEM) and transmission electron microscope (TEM). Nevertheless, the PFOS dosage of below 100â¯mg/L did not show a considerable damage to the growth of P. chrysosporium. The degradation of malachite green (MG) and 2,4-dichlorophenol (2,4-DCP) by P. chrysosporium was negatively affected by PFOS. At the initial dosage of 100â¯mg/L PFOS, the decolorization efficiency of MG and the degradation efficiency of 2,4-DCP decreased by 37% and 20%, respectively. This might be attributed to the inhibition of PFOS on MnP and LiP activities. The activities of MnP and LiP decreased by 20.6% and 43.4%, respectively. At a high dosage PFOS (100â¯mg/L), P. chrysosporium could show a high adsorption of MG but lose its pollutant degradation ability. Transcriptome analysis indicated that PFOS contamination could lead to the change of gene expression in the studied white rot fungus, and the genes regulating membrane structure, cell redox process, and cell transport, synthesis and metabolism were impacted. Membrane damage and oxidative damage were the two main mechanisms of PFOS' toxicity to P. chrysosporium.
Asunto(s)
Ácidos Alcanesulfónicos/toxicidad , Contaminantes Ambientales/metabolismo , Contaminantes Ambientales/toxicidad , Fluorocarburos/toxicidad , Phanerochaete/efectos de los fármacos , Transcriptoma/efectos de los fármacos , Adsorción , Biomasa , Clorofenoles/metabolismo , Colorantes/metabolismo , Phanerochaete/genética , Phanerochaete/crecimiento & desarrollo , Phanerochaete/metabolismo , Colorantes de Rosanilina/metabolismoRESUMEN
Hydrogen sulfide (H2S), an important cellular signaling molecule, plays vital roles in mediating responses to biotic/abiotic stresses. Influences of H2S on metal removal, cell viability, and antioxidant response of Phanerochaete chrysosporium upon exposure to heavy metals and silver nanoparticles (AgNPs) in the present study were investigated. An enhancement in Pb(ΙΙ) removal with an increase in concentration of the H2S donor sodium hydrosulfide (NaHS) was observed, and the maximum removal efficiencies increased by 31% and 17% under 100 and 200â¯mg/L Pb(ΙΙ) exposure, respectively, in the presence of 500⯵M NaHS. Application of 500⯵M NaHS increased the cell viability by 15%-39% under Pb(II) stress (10-200â¯mg/L) with relative to the untreated control. Increase in total Ag uptake and cell survival was also elicited by NaHS in a concentration-dependent manner under AgNP stress. Meanwhile, activities of superoxide dismutase and catalase were significantly enhanced with the introduction of NaHS under stresses of Pb(II), Cd(II), Cu(II), Zn(II), Ni(II), and AgNPs. The inhibition in lipid peroxidation and oxidative stress was observed in P. chrysosporium cells exposed to these toxicants following NaHS pretreatment, which could be attributed to the upregulation in antioxidant enzymes. The results obtained suggest that H2S can alleviate heavy metals and AgNP-induced toxicity to P. chrysosporium and improve the removal efficiency of these toxicants from wastewater.
Asunto(s)
Sulfuro de Hidrógeno/química , Nanopartículas del Metal/análisis , Metales Pesados/metabolismo , Phanerochaete/metabolismo , Plata/química , Antioxidantes/metabolismo , Peroxidación de Lípido , Nanopartículas del Metal/química , Nanopartículas del Metal/toxicidad , Metales Pesados/aislamiento & purificación , Metales Pesados/toxicidad , Estrés Oxidativo/efectos de los fármacos , Phanerochaete/efectos de los fármacos , Phanerochaete/crecimiento & desarrolloRESUMEN
White-rot fungi are microorganisms capable of ethanol fermentation; however, the specific conditions activating ethanol fermentation are unclear in contrast to fermentation by yeasts. In this study, we investigated the conditions favoring ethanol fermentation by the white-rot fungus Phanerochaete sordida YK-624, which is able to produce ethanol from woody material. In aerobic stationary cultivation with various concentrations of glucose (0.8-33 g/l), the fungus produced ethanol in media containing an initial glucose concentration of 2.8 g/l or higher. The amount of glucose consumption, mycelial weight, and ethanol production on the second day of culture increased in a concentration-dependent manner at low glucose concentrations; however, these were saturated at high concentrations. Biomass yields (growth/glucose consumption) were decreased until the initial glucose concentration increased to 6.0 g/l, after which the biomass yields showed constant values at higher concentrations (12-33 g/l). On the other hand, ethanol yields increased with decreasing biomass yields. In short shaking cultivation using mycelial suspension, trace amounts of instantaneous aerobic ethanol production were observed with 1.1 and 2.1 g/l glucose, but the relative gene expression levels of key enzymes at the pyruvate branch point showed no significant differences between ethanol production and non-production conditions. From these experimental results, it appears that the white-rot fungus P. sordida YK-624 produces ethanol due to overflow in sugar metabolism under aerobic conditions, although P. sordida YK-624 prioritizes glucose utilization for respiratory growth.
Asunto(s)
Etanol/metabolismo , Glucosa/metabolismo , Phanerochaete/metabolismo , Aerobiosis , Biomasa , Reactores Biológicos , Metabolismo de los Hidratos de Carbono , Medios de Cultivo/química , Fermentación , Glucosa/química , Micelio/crecimiento & desarrollo , Micelio/metabolismo , Phanerochaete/crecimiento & desarrolloRESUMEN
Ligninolytic enzyme production and lignin degradation are typically the rate-limiting steps in the biofuel industry. To improve the efficiency of simultaneous bio-delignification and enzyme production, Phanerochaete chrysosporium was transformed by shock wave-induced acoustic cavitation to co-overexpress 3 peroxidases and 1 laccase and test it on the degradation of sugarcane bagasse and wheat bran. Lignin depolymerization was enhanced by up to 25% in the presence of recombinant fungi in comparison with the wild-type strain. Sugar release on lignocellulose was 2- to 6-fold higher by recombinant fungi as compared with the control. Wheat bran ostensibly stimulated the production of ligninolytic enzymes. The highest peroxidase activity from the recombinant strains was 2.6-fold higher, whereas the increase in laccase activity was 4-fold higher in comparison to the control. The improvement of lignin degradation was directly proportional to the highest peroxidase and laccase activity. Because various phenolic compounds released during lignocellulose degradation have proven to be toxic to cells and to inhibit enzyme activity, a significant reduction (over 40%) of the total phenolic content in the samples treated with recombinant strains was observed. To our knowledge, this is the first report that engineering P. chrysosporium enhances biodegradation of lignocellulosic biomass.
Asunto(s)
Biomasa , Lacasa/biosíntesis , Lacasa/genética , Peroxidasas/biosíntesis , Peroxidasas/genética , Phanerochaete/genética , Phanerochaete/metabolismo , Biodegradación Ambiental , Biocombustibles , Celulosa/metabolismo , Clonación Molecular , Fibras de la Dieta , Ergosterol , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación Fúngica de la Expresión Génica , Lignina/metabolismo , Ingeniería Metabólica , Phanerochaete/enzimología , Phanerochaete/crecimiento & desarrollo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharum , Transformación GenéticaRESUMEN
Since the determination of the fermentation kinetics is one of the main challenges in solid state fermentation, the quantitative measurement of biomass growth during microbial pretreatment by FTIR spectroscopy in Attenuated Total Reflectance mode was evaluated. Peaks at wave numbers of 1651â¯cm-1 and 1593â¯cm-1 showed to be affected during pretreatment of poplar wood particles by Phanerochaete chrysosporium MUCL 19343. Samples with different microbial biomass fractions were obtained from two different experiments, i.e., shake flask and fixed-bed reactor experiments. The glucosamine concentration was compared to the normalized absorbance ratio of the 1651â¯cm-1 to 1593â¯cm-1 peak, measured by FTIR-ATR, and resulted in a linear relationship. The application of a normalized absorbance ratio in function of time provided a graph that was similar to the microbial growth curve. Application of FTIR in ATR mode to follow-up kinetics during solid state fermentation seems to be a fast and easy alternative to laborious measurement techniques, such as glucosamine determination.
Asunto(s)
Phanerochaete/crecimiento & desarrollo , Populus/microbiología , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Técnicas de Cultivo Celular por Lotes , Reactores Biológicos , Pared Celular/efectos de los fármacos , Quitina/análisis , Quitina/metabolismo , Glucosamina/análisis , Glucosamina/metabolismo , Cinética , Lignina/análisis , Lignina/metabolismo , Pentanonas/farmacología , Phanerochaete/efectos de los fármacos , Ácidos Sulfúricos/farmacologíaRESUMEN
The present work investigated the effect of lead (Pb) on the growth, metal accumulation, oxidative stress, and antioxidant response in Phanerochaete chrysosporium, which is a well-known hyperaccumulating species for heavy metal with appreciable bioaccumulation capacity. Results revealed that P. chrysosporium exhibited a good ability in Pb accumulation and tolerance over a concentration range of 50-100 mg L-1 Pb. The removal rate of Pb decreased with the increasing levels of Pb and reached a maximum of 91.3% at 50 mg L-1. Both extracellular adsorption and intracellular bioaccumulation contributed to the removal of Pb, with the maximum of 123.8 mg g-1 and 162.5 mg g-1 dry weight, respectively. Pb may exert its toxicity to P. chrysosporium by impairing oxidative metabolism, as evidenced by the enhanced accumulation of hydrogen peroxide (H2O2) and lipid peroxidation product malonaldehyde (MDA). P. chrysosporium evolved an antioxidant system by elevating the activity of superoxide dismutase (SOD) and the level of reduced glutathione (GSH) in response to Pb stress, whereas decreasing the activities of catalase (CAT) and peroxidase (POD). Moreover, Pearson correlation analysis demonstrated a good correlation between oxidative stress biomarkers and enzymatic antioxidants. The preset work suggested that P. chrysosporium exhibited an outstanding accumulation of Pb and tolerance of Pb-induced oxidative stress by the effective antioxidant defense mechanism.
Asunto(s)
Antioxidantes/metabolismo , Tolerancia a Medicamentos , Plomo/metabolismo , Estrés Oxidativo/efectos de los fármacos , Phanerochaete/efectos de los fármacos , Peróxido de Hidrógeno/metabolismo , Plomo/farmacología , Peroxidación de Lípido , Malondialdehído/metabolismo , Phanerochaete/enzimología , Phanerochaete/crecimiento & desarrollo , Phanerochaete/metabolismoRESUMEN
Most existing methods for screening the activity of lytic polysaccharide mono-oxygenases (LPMOs) on polysaccharides are based on the detection of soluble oxidized sugars. This approach might underestimate the total performance of LPMOs since oxidation events that do not lead to oligosaccharide release are not detected. Using PcLPMO9D as a model enzyme, a microplate-based method has been developed to detect C1-oxidizing LPMO activity by covalently linking a water-soluble fluorophore to oxidized positions within the cellulose fiber. This fluorescence method was validated using X-ray photoelectron spectroscopy and then combined with high-performance anion-exchange chromatography to track total PcLPMO9D activity.
Asunto(s)
Fluorescencia , Microtecnología/métodos , Oxigenasas de Función Mixta/metabolismo , Phanerochaete/enzimología , Polisacáridos/química , Celulosa/química , Quitina/química , Oxidación-Reducción , Phanerochaete/crecimiento & desarrollo , Espectroscopía de Fotoelectrones , Especificidad por SustratoRESUMEN
Seven white rot fungal species were tested for growth as mycelia using swine wastewater (SW), an agro-waste with tremendous environmental footprint, as the sole nutrient source. The SW contained high concentrations of carbon and nitrogen components, which could support nutritional requirements for mycelial growth. Out of the seven species, Pleurotus ostreatus and Hericium erinaceus were successfully cultivated on the SW medium using solid-state fermentation. Response surface methodology was employed to determine the combination of pH, temperature (T), and substrate concentration (C) that maximizes mycelial growth rate (Kr) for the two species. The optimum condition was estimated as pH = 5.8, T = 28.8 °C, and C = 11.2 g chemical oxygen demand (COD)/L for P. ostreatus to yield Kr of 11.0 mm/day, whereas the greatest Kr (3.1 mm/day) was anticipated at pH = 4.6, T = 25.5 °C, and C = 11.9 g COD/L for H. erinaceus. These Kr values were comparable to growth rates obtained using other substrates in the literature. These results demonstrate that SW can be used as an effective substrate for mycelial cultivation of the two white rot fungal species, suggesting an alternative method to manage SW with the production of potentially valuable biomass.
Asunto(s)
Técnicas de Cultivo Celular por Lotes/métodos , Micelio/crecimiento & desarrollo , Phanerochaete/crecimiento & desarrollo , Phanerochaete/aislamiento & purificación , Porcinos/microbiología , Aguas Residuales/microbiología , Animales , Biodegradación Ambiental , Reactores Biológicos/microbiologíaRESUMEN
With the wide production and applications of graphene and its derivatives, their toxicity to the environment has received much attention nowadays. In this study, we investigated the toxicity of graphene oxide (GO) to white rot fungus (Phanerochaete chrysosporium). GO was prepared by modified Hummers method and well characterized before use. P. chrysosporium was exposed to GO at the concentrations of 0-4 mg/mL for 7 d. The fresh and dry weights, pH values of culture media, structures, ultrastructures, IR spectra and activities of the decomposition of pollutants were measured to reveal the hazards of GO to P. chrysosporium. Our results indicated that low concentrations of GO stimulated the growth of P. chrysosporium. The exposure to GO induced more acidic pH values of the culture media after 7 d. GO induced the disruption of the fiber structure of P. chrysosporium, while at 4 mg/mL some very long and thick fibers were formed. Such changes were reflected in the scanning electron microscopy investigations, where the disruption of fibers was observed. In the ultrastructural investigations, the shape of P. chrysosporium cells changed and more vesicles were found upon the exposure to GO. The infrared spectroscopy analyses suggested that the chemical compositions of mycelia were not changed qualitatively. Beyond the toxicity, GO did not alter the activities of P. chrysosporium at low concentrations, but led to the complete loss of activity at high concentrations. The implication to the ecological safety of graphene is discussed.
Asunto(s)
Contaminantes Ambientales/toxicidad , Grafito/toxicidad , Nanoestructuras/toxicidad , Phanerochaete/efectos de los fármacos , Medios de Cultivo/química , Relación Dosis-Respuesta a Droga , Microscopía Electrónica de Rastreo , Micelio/efectos de los fármacos , Micelio/ultraestructura , Óxidos/toxicidad , Phanerochaete/crecimiento & desarrollo , Phanerochaete/ultraestructura , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Due to the particular activation and inhibition behavior of silver nanoparticles (AgNPs) on microbes at various concentrations, it's crucial to exploit the special concentration effect in environment. Here, we studied the viability variation of Phanerochaete chrysosporium (P. chrysosporium) under exposure to citrate-coated AgNPs (Citrate-AgNPs) in the presence of different sulfide sources (an inorganic sulfide, NaHS and an organic sulfide, thioacetamide (TAA)). The results indicated that both NaHS and TAA can promote activation of P. chrysosporium by Citrate-AgNPs at a higher concentration, which was initial at toxic level. Treatment with various concentrations of Citrate-AgNPs (0-9 mg/L) demonstrated a maximum activation concentration (MAC) at 3 mg/L. With the increase in sulfide concentration, MAC transferred to higher concentration significantly, indicating the obvious "toxicity to activation" transformation at a higher concentration. Ag(+) testing exhibited that variations in sulfide-induced Ag(+) concentration (3-7 µg/L Ag(+)) accounted for the "toxicity to activation" transformation. In addition, the similar results were observed on antibacterial application using Escherichia coli as the model species. Based on the research results, the application of this transformation in improving antibacterial activity was proposed. Therefore, the antibacterial activity of AgNPs can be controlled, even at concentration, via adjusting for the sulfide concentration.
Asunto(s)
Antibacterianos/farmacología , Nanopartículas del Metal/química , Phanerochaete/efectos de los fármacos , Plata/farmacología , Sulfuros/farmacología , Tioacetamida/farmacología , Ácido Cítrico/farmacología , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Sinergismo Farmacológico , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Phanerochaete/crecimiento & desarrolloRESUMEN
This study examines the growth, metabolism of Phanerochaete chrysosporium (P. chrysosporium) and route of lignin degradation in response to cadmium (Cd) stress in solid-state fermentation of rice straw. Less living fungi biomass was found under Cd exposure, suggesting that Cd had strong toxicity to P. chrysosporium. The maximum values of lignin peroxidase and manganese peroxidase were 0.34 and 5.21 U g(-1) at the Cd concentration of 32 mg kg(-1), respectively, lower than that in control, which indicated Cd stress would inhibit ligninolytic enzymes. The production of reactive oxygen species (ROS) including hydroxyl radicals (OH), superoxide anion radical (O2(-)) and hydrogen peroxide (H2O2) increased after Cd exposure. Higher concentration of oxalate was detected at high Cd concentrations. Cd stress also had influence on the rates of lignocelluloses degradation and the route of lignin degradation. Partial Cd could be removed by P. chrysosporium.