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1.
Int J Mol Sci ; 25(11)2024 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-38891868

RESUMO

Mycoviruses are usually transmitted horizontally via hyphal anastomosis and vertically through sporulation in natural settings. Oyster mushroom spherical virus (OMSV) is a mycovirus that infects Pleurotus ostreatus, with horizontal transmission via hyphal anastomosis. However, whether OMSV can be vertically transmitted is unclear. This study aimed to investigate the transmission characteristics of OMSV to progeny via basidiospores and horizontally to a new host. A total of 37 single-basidiospore offspring were obtained from OMSV-infected P. ostreatus and Pleurotus pulmonarius for Western blot detection of OMSV. The OMSV-carrying rate among monokaryotic isolates was 19% in P. ostreatus and 44% in P. pulmonarius. Then, OMSV-free and OMSV-infected monokaryotic isolates were selected for hybridization with harvested dikaryotic progeny strains. Western blot analyses of the offspring revealed that the OMSV transmission efficiency was 50% in P. ostreatus and 75% in P. pulmonarius, indicating vertical transmission via sexual basidiospores. Furthermore, we observed the horizontal transfer of OMSV from P. pulmonarius to Pleurotus floridanus. OMSV infection in P. floridanus resulted in significant inhibition of mycelial growth and yield loss. This study was novel in reporting the vertical transmission of OMSV through basidiospores, and its infection and pathogenicity in a new host P. floridanus.


Assuntos
Micovírus , Pleurotus , Esporos Fúngicos , Pleurotus/virologia , Esporos Fúngicos/crescimento & desenvolvimento , Micovírus/fisiologia
2.
Biotechnol Appl Biochem ; 70(1): 210-220, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35398919

RESUMO

Recently, composting cultivation method is widely used in oyster mushroom production. In this study, we focused on the effects of composting processes on nutritional qualities and antioxidant activity of Pleurotus floridanus mushroom fruiting bodies. Three treatments of different composting time (2, 4, and 5 days) were performed with an atmospheric sterilization treatment as the control. The results showed that the pH value, total carbon content, and total nitrogen content of substrate were critical parameters which would significantly affect mushroom qualities and bioactivities. Fruiting bodies of the control demonstrated significantly higher crude protein content, total amino acid content, and essential amino acid content than that of composting treatments. Moreover, fruiting bodies of treatment D4 and D5 manifested significantly higher crude polysaccharide contents. Crude polysaccharide of treatment D4 represented the highest scavenging ability toward both radical 3-ethylbenzthiazoline-6-sulfonic acid (ABTS·+ ) and Hydroxyl radical (OH·). It suggests that composting processes is suitable for oyster mushroom cultivation based on nutritional and antioxidant qualities of fruiting bodies.


Assuntos
Compostagem , Pleurotus , Prunus persica , Antioxidantes/química , Pleurotus/metabolismo
3.
Proc Natl Acad Sci U S A ; 117(11): 6014-6022, 2020 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-32123065

RESUMO

Fungal predatory behavior on nematodes has evolved independently in all major fungal lineages. The basidiomycete oyster mushroom Pleurotus ostreatus is a carnivorous fungus that preys on nematodes to supplement its nitrogen intake under nutrient-limiting conditions. Its hyphae can paralyze nematodes within a few minutes of contact, but the mechanism had remained unclear. We demonstrate that the predator-prey relationship is highly conserved between multiple Pleurotus species and a diversity of nematodes. To further investigate the cellular and molecular mechanisms underlying rapid nematode paralysis, we conducted genetic screens in Caenorhabditis elegans and isolated mutants that became resistant to P. ostreatus We found that paralysis-resistant mutants all harbored loss-of-function mutations in genes required for ciliogenesis, demonstrating that the fungus induced paralysis via the cilia of nematode sensory neurons. Furthermore, we observed that P. ostreatus caused excess calcium influx and hypercontraction of the head and pharyngeal muscle cells, ultimately resulting in rapid necrosis of the entire nervous system and muscle cells throughout the entire organism. This cilia-dependent predatory mechanism is evolutionarily conserved in Pristionchus pacificus, a nematode species estimated to have diverged from C. elegans 280 to 430 million y ago. Thus, P. ostreatus exploits a nematode-killing mechanism that is distinct from widely used anthelmintic drugs such as ivermectin, levamisole, and aldicarb, representing a potential route for targeting parasitic nematodes in plants, animals, and humans.


Assuntos
Caenorhabditis elegans/efeitos dos fármacos , Micotoxinas/toxicidade , Pleurotus/fisiologia , Animais , Caenorhabditis elegans/citologia , Caenorhabditis elegans/fisiologia , Cálcio/metabolismo , Cílios/fisiologia , Músculo Esquelético/citologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/inervação , Necrose/induzido quimicamente
4.
Chem Biodivers ; 20(9): e202300346, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37503864

RESUMO

Pleurotus ostreatus is an edible fungus with high nutritional value that uses industrial and agricultural lignocellulosic residues as substrates for growth and reproduction. Understanding their growth metabolic dynamics on agro-industrial wastes would help to develop economically viable and eco-friendly biotechnological strategies for food production. Thus, we used UHPLC/MS/MS and GNPS as an innovative approach to investigate the chemical composition of two strains of P. ostreatus, coded as BH (Black Hirataki) and WH (White Hirataki), grown on sisal waste mixture (SW) supplemented with 20 % cocoa almond tegument (CAT) or 20 % of wheat bran (WB). Metabolite dereplication allowed the identification of 53 metabolites, which included glycerophospholipids, fatty acids, monoacylglycerols, steroids, carbohydrates, amino acids, and flavonoids. This is the first report of the identification of these compounds in P. ostreatus, except for the steroid ergosterol. Most of the metabolites described in this work possess potential biological activities, which support the nutraceutical properties of P. ostreatus. Thus, the results of this study provide essential leads to the understanding of white-rot fungi chemical plasticity aiming at developing alternative biotechnologies strategies for waste recycling.


Assuntos
Pleurotus , Prunus dulcis , Pleurotus/química , Pleurotus/metabolismo , Resíduos Industriais , Fibras na Dieta/metabolismo , Espectrometria de Massas em Tandem , Suplementos Nutricionais
5.
Int J Mol Sci ; 24(13)2023 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-37445762

RESUMO

Oyster mushroom spherical virus (OMSV) is a mycovirus with a positive-sense single-stranded RNA genome that infects the edible mushroom Pleurotus ostreatus. OMSV is horizontally transferred from an infected strain to a cured strain via mycelia. The infection results in significant inhibition of mycelial growth, malformation of fruiting bodies, and yield loss in oyster mushrooms. This study successfully transferred OMSV from P. ostreatus to Pleurotus pulmonarius. However, transmission was not successful in other Pleurotus species including P. citrinopileatus, P. eryngii, P. nebrodensis, and P. salmoneostramineus. The successful OMSV infection in P. pulmonarius was further verified with Western blot analysis using a newly prepared polyclonal antiserum against the OMSV coat protein. Furthermore, OMSV infection reduced the mycelial growth rate of P. pulmonarius. The OMSV-infected strain demonstrated abnormal performance including twisted mushrooms or irregular edge of the cap as well as reduced yield of fruiting bodies in P. pulmonarius, compared to the OMSV-free strain. This study is the first report on the infection and pathogenicity of OMSV to the new host P. pulmonarius. The data from this study therefore suggest that OMSV is a potential threat to P. pulmonarius.


Assuntos
Micovírus , Pleurotus , Vírus de RNA , Pleurotus/genética , Vírus de RNA/genética
6.
Molecules ; 28(13)2023 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-37446941

RESUMO

When added to mushroom growing substrates, edible and medicinal herbs affect the mushrooms' nutritional and medicinal value. In this study, polysaccharides (P0OP-I and P15OP-I) were extracted and purified from oyster mushrooms grown on substrates supplemented with 0% and 15% Astragalus roots (P0 and P15), respectively, and their chemical structure and immunobiological activities were compared. P15OP-I and P0OP-I were extracted using ultrasound-assisted hot water and deproteinized with the Sevage method, depigmented with 30% H2O2, desalted with dialysis, and purified using DEAE-52 cellulose and Sephadex G-100 dextran column chromatography. The molecular weight of P0OP-I and P15OP-I was 21,706.96 and 20,172.65 Da, respectively. Both were composed of monosaccharides D-mannose, galacturonic acid, D-glucose, D-galactose, and L-arabinose but in different molar ratios, and both were connected by a pyranoside linkage. P15OP-I consisted of higher contents of mannose, glucose, galactose and arabinose and lower content of galacturonic acid as compared to P0OP-I. Both P0OP-I and P15OP-I induced NO and TNF-α production but did not show cytotoxic effect or induce ROS generation in RAW264.7 cells. P15OP-I showed a stronger ability to promote NO and TNF-α production relative to P0OP-I. In vitro experiments showed that the immunomodulatory activity of P0OP-I and P15OP-I in RAW264.7 macrophages were mediated by the JNK/MAPK, Erk/MAPK, and NF-κB signaling pathways. The results would be helpful for elucidation of the health promoting mechanism of Astragalus oyster mushrooms as a source of neutraceuticals.


Assuntos
Astrágalo , Pleurotus , Pleurotus/química , Fator de Necrose Tumoral alfa , Peróxido de Hidrogênio , Diálise Renal , Polissacarídeos/farmacologia , Polissacarídeos/química , Astrágalo/química
7.
Molecules ; 28(19)2023 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-37836819

RESUMO

The growing interest in functional food makes looking for new possibilities of enriching products with health-promoting ingredients necessary. One raw material with a very high potential for the food industry is the oyster mushroom (Pleurotus ostreatus), which has a strong antioxidant, antiviral, and anticancer effect. Carp meat (Cyprinus carpio) also has beneficial properties. It is rich in easily digestible protein, vitamins, minerals, and omega-3 fatty acids. This study aimed to evaluate the effect of oyster mushroom addition on the quality of carp burgers, with particular emphasis on the antioxidant effect. The scientific literature produced so far has not focused on the synergy between oyster mushrooms and carp meat. The addition of oyster mushrooms contributed to the increase in antioxidant properties and sensory attractiveness of burgers. The fat content in the finished product was reduced, and the degree of their oxidation was also reduced. The obtained results will contribute to the creation of innovative food products that meet the expectations of consumers looking for healthy food.


Assuntos
Carpas , Pleurotus , Animais , Antioxidantes/farmacologia , Carne , Valor Nutritivo
8.
Curr Issues Mol Biol ; 44(11): 5778-5787, 2022 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-36421676

RESUMO

Mycoviruses are viruses that specifically infect and replicate in fungi. Several mycoviruses have been previously reported in Pleurotus ostreatus, including the oyster mushroom spherical virus (OMSV), oyster mushroom isometric virus (OMIV), Pleurotus ostreatus spherical virus (POSV), and Pleurotus ostreatus virus 1 (PoV1). This study was designed to develop a multiplex RT-PCR for simultaneous detection and differentiation of the four P. ostreatus mycoviruses. Four pairs of primers were designed from conserved regions based on the reported sequences and the multiplex RT-PCR products were 672 bp for OMSV, 540 bp for OMIV, 310 bp for POSV, and 200 bp for PoV1. The optimal annealing temperature of the multiplex RT-PCR was 62 °C and the detection limits of the plasmids were 100 fg for OMSV and OMIV and 1 pg for POSV and PoV1. This technique was successfully applied for the detection of OMSV, OMIV, and POSV from different P. ostreatus strains and the plasmid containing the PoV1 sequence. This methodology can serve as a powerful diagnostic tool for the survey of the incidence and epidemiology of the four P. ostreatus mycoviruses, further contributing to the prevention and treatment of mycoviral diseases in P. ostreatus.

9.
Fish Shellfish Immunol ; 120: 155-165, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34822996

RESUMO

This study aimed to evaluate the effects of a synbiotic composite an extract from a by-product of king oyster mushroom, Pleurotus eryngii (KOME), and probiotic Lactobacillus plantarum 7-40 on the growth performance and health status of white shrimp, Litopenaeus vannamei. The KOME was able to stimulate the growth of probiotic, but not the growth of Vibrio pathogens, including V. alginolyticus, V. parahaemolyticus, and V. harveyi. Four diets were formulated, including a control diet supplemented without prebiotic and probiotic, a basal diet supplemented with KOME (5 g kg-1) (ME), a basal diet supplemented with probiotic (1 × 108 CFU kg-1) (LP), and a basal diet supplemented with KOME (5 g kg-1) and probiotic (1 × 108 CFU kg-1) (SYN). Shrimp fed the ME, LP, and SYN diets had significantly higher survival than that of shrimp fed with the control diet for 8 weeks. Shrimp in the SYN group also had a significantly higher weight gain and total final weight in comparison with the control and other treatments. In the intestinal tract, lactic acid bacteria count was significantly higher in the SYN group, whereas the Vibrio-like bacteria count was significantly higher in the ME group than in the control group. For the health status assessment, the disease resistance of shrimp against V. alginolyticus was improved in all treatments compared to the shrimp in control. Shrimps in the SYN group had significantly lower cumulative mortality due to the significant increase in immune responses, including phenoloxidase, respiratory burst, and lysozyme activity, and the gene expression of pexn and pen4 in the haemocytes, and lgbp, sp, propoii, pexn, pen3a, pen4, and gpx in the haepatopancreas of shrimp as compared to the control. Therefore, it is suggested that a combination of KOME and probiotics can be used as a synbiotic to improve the growth performance and reduce the risk of infectious diseases caused by Vibrio and at the same time significantly contribute to the circular economy.


Assuntos
Lactobacillus plantarum , Penaeidae , Pleurotus , Probióticos , Simbióticos , Animais , Dieta/veterinária , Imunidade Inata , Penaeidae/crescimento & desenvolvimento , Pleurotus/química , Prebióticos , Vibrio
10.
J Appl Microbiol ; 132(3): 2131-2143, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34637602

RESUMO

AIMS: Green-mould contamination is identified as one of the challenges faced by mushroom cultivation industry globally which believed to be caused by Trichoderma spp. METHODS AND RESULTS: To explore the dynamics of microbial population in mushroom substrate during commercial mushroom cultivation and how microbiota might play a role in green-mould contamination, we applied both culturing and targeted metagenomics approaches to identify microbiota in noncomposted sawdust substrates at different cultivation stages. The microbiological analysis showed that the green-mould contaminated substrates harboured higher total mesophilic bacteria count. The green-moulds isolated from the contaminated mushroom substrates were identified as Trichoderma pleurotum (n = 15; 93.8%) and Graphium penicillioides (n = 1; 6.3%). To our surprise, the targeted metagenomic analysis revealed that Graphium comprised 56.3% while Trichoderma consisted of only 36.1% of the total fungi population, suggesting that green-mould contamination might not be caused by Trichoderma alone, but also Graphium that grows very slowly in the laboratory. CONCLUSION: It is worthwhile to note that G. penicillioides was also isolated in the early stages of mushroom cultivation, but not T. pleurotum. The results indicated that the structure and composition of the bacterial population in the mushroom substrate varied and the bacterial population shifted along the cultivation process. SIGNIFICANCE AND IMPACT OF STUDY: This study revealed a possibility of G. penicillioides as an overlooked fungi causing green-mould contamination.


Assuntos
Agaricales , Microbiota , Pleurotus , Bactérias/genética
11.
Lett Appl Microbiol ; 74(1): 44-52, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34614248

RESUMO

We investigated whether highly available organic residues in Brazil can be used as substrates for the production of the oyster mushroom Pleurotus ostreatus, instead of the conventional cultivation using the eucalyptus sawdust substrate. We assessed the mushroom yield on 13 substrates, of which 12 were formulated with different concentrations of organic residues and one with pure eucalyptus sawdust, and verified whether the raw material used in the substrate formula and the concentration of such alternative residues influenced their biological efficiencies. Substrates containing eucalyptus bark resulted in higher mushroom yield than those containing eucalyptus sawdust, which generally resulted in similar mushroom yield to the remaining formulas. Moreover, the raw material and the concentration of each residue affected the biological efficiency of the substrates. We show that the conventional substrate for P. ostreatus can be replaced by substrates easily accessible to producers without loss in productivity. Furthermore, that the concentration of these mixtures affects the mushroom productivity and should be considered when formulating the growth medium.


Assuntos
Agaricales , Pleurotus , Agricultura , Brasil , Florestas
12.
Anal Biochem ; 635: 114450, 2021 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-34767809

RESUMO

Lectin is a carbohydrate-binding protein, which exhibits a plethora of biological properties such as antimicrobial, antifungal, and anticancer activities. In the present study, lectin, with an antibacterial and antioxidant potential, was purified from the oyster mushroom Pleurotus flabellatus. The P. flabellatus Lectin (PFL-L) was purified by using a DEAE - cellulose anion exchange chromatography followed by gel-filtration chromatography. The PFL-L was characterized by CD, HPLC, and MALDI-TOF/MS. The purity of PFL-L increased to 62.40% with the recovery of hemagglutinating activity (HA) by 12.12%. On SDS - PAGE, the PFL-L gave a single band of 18 kDa. PFL-L, consisting of d-galactose, exhibits a strong hemagglutinating activity. It was stable at pH (6.0-7.5) and temperature (10-20 °C) in addition to having extensive hemagglutinating activity. PFL-L enhanced the HA with the use of different metal ions namely Mg2+, Ca2+, and Fe2+. The study of bacterial growth inhibition led to the inference that the PFL-L was more potent against gram-negative bacteria. PFL-L showed the highest radical scavenging activity for the DPPH assay at 100 µg/mL (89.9 ± 2.53%). The highest antioxidant activities with IC50 values (for DPPH assay) of 53.96 µg/mL were determined for PFL-L and the present study shows that lectin from P. flabellatus manifested distinctive character and potentially exploitable activities.


Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Lectinas/farmacologia , Pleurotus/química , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Compostos de Bifenilo/antagonistas & inibidores , Lectinas/química , Lectinas/isolamento & purificação , Testes de Sensibilidade Microbiana , Picratos/antagonistas & inibidores , Ovinos
13.
Amino Acids ; 53(7): 1105-1122, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34106335

RESUMO

Free amino acids (FAAs), the major constituents of the natural moisturizing factor (NMF), are very important for maintaining the moisture balance of human skin and their deficiency results in dry skin conditions. There is a great interest in the identification and use of nature-based sources of these molecules for such cosmeceutical applications. The objective of the present study was, therefore, to investigate the FAA contents of selected Ethiopian plant and fungi species; and select the best sources so as to use them for the stated purpose. About 59 different plant species and oyster mushroom were included in the study and the concentrations of 27 FAAs were analyzed. Each sample was collected, lyophilized, extracted using aqueous solvent, derivatized with Fluorenylmethoxycarbonyl chloride (Fmoc-Cl) prior to solid-phase extraction and quantified using Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC-ESI-MS/MS) system. All the 27 FAAs were detected in most of the samples. The dominant FAAs that are part of the NMF were found at sufficiently high concentration in the mushroom and some of the plants. This indicates that FAAs that could be included in the preparations for the management of dry skin condition can be obtained from a single natural resource and the use of these resources for the specified purpose have both economic and therapeutic advantage in addition to fulfilling customer needs.


Assuntos
Aminoácidos/metabolismo , Cosmecêuticos/metabolismo , Fungos/metabolismo , Plantas/metabolismo , Pele/metabolismo , Aminoácidos/análise , Cromatografia Líquida , Cosmecêuticos/análise , Humanos , Espectrometria de Massas em Tandem
14.
J Invertebr Pathol ; 186: 107474, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-32971130

RESUMO

The aegerolysin proteins ostreolysin A6, pleurotolysin A2 and erylysin A are produced by mushrooms of the genus Pleurotus. These aegerolysins can interact specifically with sphingolipid-enriched membranes. In particular, they strongly bind insect cells and to artificial lipid membranes that contain physiologically relevant concentrations of the main invertebrate-specific sphingolipid, ceramide phosphoethanolamine. Moreover, the aegerolysins permeabilise these membranes when combined with their protein partner pleurotolysin B, which contains a membrane-attack-complex/perforin domain. These aegerolysin/ pleurotolysin B complexes show strong and selective toxicity towards western corn rootworm larvae and adults and Colorado potato beetle larvae. Their insecticidal activities arise through aegerolysin binding to ceramide phosphoethanolamine in the insect midgut. This mode of membrane binding is different from those described for similar aegerolysin-based complexes of bacterial origin (e.g., Cry34Ab1/Cry35Ab1), or other Bacillus thuringiensis proteinaceous crystal toxins, which associate with protein receptors. The ability of Pleurotus aegerolysins to specifically interact with sphingolipid-enriched domains in mammalian cells can be further exploited to visualize lipid rafts in living cells, and to treat certain types of tumours and metabolic disorders. Finally, these proteins can strongly enhance fruiting initiation of P. ostreatus even when applied externally. In this review, we summarise the current knowledge of the potential biotechnological and biomedical applications of the Pleurotus aegerolysins, either alone or when complexed with pleurotolysin B, with special emphasis on their bioinsecticidal effects.


Assuntos
Besouros/efeitos dos fármacos , Proteínas Fúngicas/farmacologia , Proteínas Hemolisinas/farmacologia , Inseticidas/farmacologia , Controle Biológico de Vetores , Pleurotus/química , Animais , Agentes de Controle Biológico , Besouros/crescimento & desenvolvimento , Proteínas de Drosophila , Proteínas Fúngicas/química , Proteínas Hemolisinas/química , Inseticidas/química , Fatores de Transcrição
15.
J Environ Sci Health B ; 56(1): 54-63, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33156729

RESUMO

Twenty-two strains of Trichoderma spp. (T. harzianum species complex [THSC], Trichoderma aggressivum f. europaeum, Trichoderma pleuroti, and Trichoderma pleuroticola) causing green mold disease on edible mushrooms (button mushroom, shiitake and oyster mushroom), collected during 2004-2018 from four countries (Serbia, North Macedonia, Croatia, and Hungary) were examined. Based on their ITS (internal transcribed spacer) sequences, strains from shiitake mushroom in Serbia were identified as members of the THSC, while in samples obtained from Serbian and North-Macedonian oyster mushroom farms THSC, T. pleuroti and T. pleuroticola were detected, which represent the first findings in the region. In fungicide susceptibility tests, all examined Trichoderma strains were found to be highly sensitive to prochloraz (ED50<0.4 µg mL-1) and considerably susceptible to metrafenone (ED50 < 4 µg mL-1). The most sensitive taxon to both fungicides was THSC from oyster mushroom. The toxicity of metrafenone was satisfying and strains from oyster mushroom showed the highest sensitivity (ED50 < 1.43 µg mL-1), while strains originating from button mushroom and shiitake displayed similar susceptibilities (ED50 < 3.64 µg mL-1). After additional in vivo trials, metrafenone might also be recommended for the control of green mold disease in mushroom farms.


Assuntos
Benzofenonas/farmacologia , Fungicidas Industriais/farmacologia , Imidazóis/farmacologia , Trichoderma/efeitos dos fármacos , Agaricus/efeitos dos fármacos , Agaricus/crescimento & desenvolvimento , Europa Oriental , Testes de Sensibilidade Microbiana , Trichoderma/classificação
16.
J Environ Manage ; 272: 111087, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32669250

RESUMO

Efficient removal techniques are urgently needed to remove organic micropollutants (OMPs) from wastewater, in order to protect water resources. In this study, laccase activity of mushroom substrate colonized by Pleurotus ostreatus was evaluated as a novel wastewater treatment method for removal of OMPs, including diclofenac, bicalutamide, lamotrigine, and metformin at environmentally relevant concentrations. Laccase activity of the colonized mushroom substrate was found to be highest, 0.8 enzyme activity (U)/g mushroom substrate wet weight, immediately before initiation of fruiting body formation. The selected OMPs were treated for 5 min with suspensions of mushroom substrate with laccase activity of approximately 50 U/L. Removal of all OMPs was significant, with the highest removal for diclofenac of 90% compared with a control with uncolonized mushroom substrate. To our knowledge, direct use of colonized mushroom substrate in removing diclofenac from water has not been reported previously. Removal efficiency of bicalutamide, lamotrigine, and metformin was 43%, 73%, and 59%, respectively. This demonstrates potential for using mushroom substrate colonized by P. ostreatus for removal of OMPs from wastewater.


Assuntos
Agaricales , Pleurotus , Lignina , Água
17.
BMC Biotechnol ; 19(1): 43, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31262286

RESUMO

BACKGROUND: Proteases are hydrolytic enzymes that catalyze peptide linkage cleavage reactions at the level of proteins and peptides with different degrees of specificity. This group draws the attention of industry. More than one protease in three is a serine protease. Classically, they are active at neutral to alkaline pH. The serine proteases are researched for industrial uses, especially detergents. They are the most commercially available enzyme group in the world market. Overall, fungi produced extracellular proteases, easily separated from mycelium by filtration. RESULTS: A new basidiomycete fungus CTM10057, a hyperproducer of a novel protease (10,500 U/mL), was identified as Pleurotus sajor-caju (oyster mushroom). The enzyme, called SPPS, was purified to homogeneity by heat-treatment (80 °C for 20 min) followed by ammonium sulfate precipitation (35-55%)-dialysis, then UNO Q-6 FPLC ion-exchange chromatography and finally HPLC-ZORBAX PSM 300 HPSEC gel filtration chromatography, and submitted to biochemical characterization assays. The molecular mass was estimated to be 65 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Native-PAGE, casein-zymography, and size exclusion by HPLC. A high homology with mushroom proteases was displayed by the first 26 amino-acid residues of the NH2-terminal aminoacid sequence. Phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP) strongly inhibit SPPS, revealing that it is a member of the serine-proteases family. The pH and temperature optima were 9.5 and 70 °C, respectively. Interestingly, SPPS possesses the most elevated hydrolysis level and catalytic efficiency in comparison with SPTC, Flavourzyme® 500 L, and Thermolysin type X proteases. More remarkably, a high tolerance towards organic solvent tolerance was exhibited by SPPS, together with considerable detergent stability compared to the commercial proteases Thermolysin type X and Flavourzyme® 500 L, respectively. CONCLUSIONS: This proves the excellent proprieties characterizing SPPS, making it a potential candidate for industrial applications especially detergent formulations.


Assuntos
Proteínas Fúngicas/metabolismo , Temperatura Alta , Pleurotus/enzimologia , Serina Proteases/metabolismo , Detergentes/química , Estabilidade Enzimática , Proteínas Fúngicas/química , Proteínas Fúngicas/isolamento & purificação , Concentração de Íons de Hidrogênio , Hidrólise , Microbiologia Industrial/métodos , Cinética , Peso Molecular , Serina Proteases/química , Serina Proteases/isolamento & purificação , Especificidade por Substrato
18.
Appl Microbiol Biotechnol ; 103(14): 5831-5841, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115628

RESUMO

In coffee-producing countries, waste products from coffee production are useful substrates for cultivation of Pleurotus ostreatus. This species is relatively easy to grow, coffee waste substrates are readily available and the mushroom fruiting bodies are a valuable source of nutrition and income. In developed countries, cultivation of P. ostreatus on spent coffee grounds (SCG) from coffee consumption is a novel way to recycle this urban waste product. Here, we studied the effect of SCG and caffeine on growth of a commercial strain of P. ostreatus in liquid and solid cultures, and on a commercial scale. The presence of caffeine inhibited mycelial growth on agar and in liquid culture in the laboratory. Increased levels of SCG in an SCG/sawdust substrate also delayed mycelial growth and delayed or prevented fruiting during commercial cultivation. Despite growth inhibition, partial degradation of caffeine to xanthine by P. ostreatus mycelium was observed in all SCG-containing substrate mixtures. Degradation of caffeine proceeded mainly via sequential N-demethylation to theophylline (1,3-dimethylxanthine) and 3-methylxanthine, although both paraxanthine and theobromine also accumulated in the substrate. Caffeine and its demethylated metabolites were also detected in fruiting bodies, but it was not clear whether caffeine metabolism occurred in the fruiting bodies themselves or whether caffeine metabolites were translocated there from the mycelium. Based on the caffeine concentrations measured in fruiting bodies after growth with SCG, it would be necessary to consume ~ 250 kg of fresh oyster mushrooms to obtain the amount of caffeine equivalent to one cup of espresso coffee, suggesting that the health impact of caffeine in these mushrooms is low. However, the ability of P. ostreatus to degrade caffeine indicates that this and other species in this genus may have potential applications in detoxification of coffee production wastes.


Assuntos
Cafeína/metabolismo , Pleurotus/crescimento & desenvolvimento , Pleurotus/metabolismo , Resíduos/análise , Café/química , Meios de Cultura/química , Carpóforos/crescimento & desenvolvimento , Carpóforos/metabolismo , Microbiologia Industrial , Resíduos Industriais/análise , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Xantina/metabolismo
19.
Mikrochim Acta ; 186(8): 510, 2019 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-31280386

RESUMO

The design and preparation of magnetic cucurbit[6]uril hypercrosslinked with polymers are described. The materials have a large specific surface, abundant mesopores and cavities, and display superparamagnetism. They were applied to the enrichment of ubiquitinated peptides from standard protein digests. Following desorption with 0.15% TFA, the peptides were quantified by MALDI-TOF MS. The method has a detection limit of 2 fmol·µL-1 and a mass ratio selectivity of 1:5000 as shown for ubiquitin and bovine serum albumin. The materials enable selective capture of ubiquitinated peptides from genuine samples comprising of oyster mushroom and human serum. This demonstrates their potential for the analysis of low-level ubiquitin in complex samples. Graphical abstract Schematic presentation for the synthesis of magnetic cucurbit[6]urils hypercrosslinked polymers (MagCB[6]-HCPs).

20.
J Sci Food Agric ; 99(7): 3616-3627, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30628086

RESUMO

BACKGROUND: Pleurotus ostreatus mushroom (POM) is an edible mushroom with rich nutritional components and vital pharmacological properties. The present study comprised 100 cross-bred piglets, weaned at 28 days old, who were randomly assigned to four POM diets with five replicates per diet and five piglets per pen. RESULTS: POM supplementation (P < 0.05) decreased the incidence of diarrhea, and also increased the average daily feed intake and average daily gain of pigs. Fecal acetate, butyrate and propionate increased with the addition of POM. Interleukin-2, immunoglobulin G, immunoglobulin M, tumor necrosis factor-α and immunoglobulin A increased (P < 0.05) with the addition of POM. The 16S rDNA sequencing results showed that the Bacteroidetes and Firmicutes were the dominant microbial strains in the fecal samples, irrespective of POM supplementation. Shannon diversity, whole tree phylogenetic diversity, observed species and Chao1 analysis exhibited significant variation in species richness across the treatments. Principal coordinates analysis showed a significant (P < 0.1) increase in the microbial communities amongst all of the treatment groups. CONCLUSION: The results of the present study suggest that the supplementation of POM in the diet of piglets might increase feed consumption, gut microbial composition and diversity, as well as short-chain fatty acids synthesis, consequently preventing the occurrence of diarrhea and increasing the growth of piglets. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Ração Animal/análise , Diarreia/veterinária , Microbioma Gastrointestinal , Pleurotus/metabolismo , Doenças dos Suínos/imunologia , Suínos/crescimento & desenvolvimento , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Diarreia/imunologia , Diarreia/microbiologia , Diarreia/prevenção & controle , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Fezes , Feminino , Imunidade , Intestinos/imunologia , Intestinos/microbiologia , Masculino , Microbiota , Suínos/imunologia , Suínos/metabolismo , Suínos/microbiologia , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle
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