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1.
Molecules ; 26(15)2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-34361776

RESUMO

In this study, we examined aqueous extracts of the edible mushrooms Pleurotus ostreatus (oyster mushroom) and Lentinula edodes (shiitake mushroom). Proteome analysis was conducted using LC-Triple TOF-MS and showed the expression of 753 proteins by Pleurotus ostreatus, and 432 proteins by Lentinula edodes. Bioactive peptides: Rab GDP dissociation inhibitor, superoxide dismutase, thioredoxin reductase, serine proteinase and lectin, were identified in both mushrooms. The extracts also included promising bioactive compounds including phenolics, flavonoids, vitamins and amino acids. The extracts showed promising antiviral activities, with a selectivity index (SI) of 4.5 for Pleurotus ostreatus against adenovirus (Ad7), and a slight activity for Lentinula edodes against herpes simplex-II (HSV-2). The extracts were not cytotoxic to normal human peripheral blood mononuclear cells (PBMCs). On the contrary, they showed moderate cytotoxicity against various cancer cell lines. Additionally, antioxidant activity was assessed using DPPH radical scavenging, ABTS radical cation scavenging and ORAC assays. The two extracts showed potential antioxidant activities, with the maximum activity seen for Pleurotus ostreatus (IC50 µg/mL) = 39.46 ± 1.27 for DPPH; 11.22 ± 1.81 for ABTS; and 21.40 ± 2.20 for ORAC assays. This study encourages the use of these mushrooms in medicine in the light of their low cytotoxicity on normal PBMCs vis à vis their antiviral, antitumor and antioxidant capabilities.


Assuntos
Antineoplásicos/química , Antioxidantes/química , Antivirais/química , Proteínas Fúngicas/química , Pleurotus/química , Proteoma/química , Cogumelos Shiitake/química , Aminoácidos/química , Aminoácidos/isolamento & purificação , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Antivirais/isolamento & purificação , Antivirais/farmacologia , Benzotiazóis/antagonistas & inibidores , Compostos de Bifenilo/antagonistas & inibidores , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Misturas Complexas/química , Flavonoides/química , Flavonoides/isolamento & purificação , Proteínas Fúngicas/classificação , Proteínas Fúngicas/isolamento & purificação , Humanos , Lectinas/química , Lectinas/isolamento & purificação , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Especificidade de Órgãos , Fenóis/química , Fenóis/isolamento & purificação , Picratos/antagonistas & inibidores , Pleurotus/metabolismo , Cultura Primária de Células , Proteoma/classificação , Proteoma/isolamento & purificação , Serina Proteases/química , Serina Proteases/isolamento & purificação , Cogumelos Shiitake/metabolismo , Ácidos Sulfônicos/antagonistas & inibidores , Superóxido Dismutase/química , Superóxido Dismutase/isolamento & purificação , Tiorredoxina Dissulfeto Redutase/química , Tiorredoxina Dissulfeto Redutase/isolamento & purificação , Vitaminas/química , Vitaminas/isolamento & purificação , Água/química
2.
Nutrients ; 13(8)2021 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-34444724

RESUMO

Astragalus root (Huang Qi) and Shiitake mushrooms (Lentinus edodes) are both considered medicinal foods and are frequently used in traditional Chinese medicine due to their anticancer and immunomodulating properties. Here, the scientific literatures describing evidence for the anticancer and immunogenic properties of Shiitake and Astragalus were reviewed. Based on our experimental data, the potential to develop medicinal food with combined bioactivities was assessed using Shiitake mushrooms grown over Astragalus beds in a proprietary manufacturing process, as a novel cancer prevention approach. Notably, our data suggest that this new manufacturing process can result in transfer and increased bioavailability of Astragalus polysaccharides with therapeutic potential into edible Shiitake. Further research efforts are required to validate the therapeutic potential of this new Hengshan Astragalus Shiitake medicinal food.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Astrágalo (Planta) , Alimento Funcional , Imunomodulação , Neoplasias/terapia , Cogumelos Shiitake , Animais , Antineoplásicos Fitogênicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica , Astrágalo (Planta)/química , Astrágalo (Planta)/crescimento & desenvolvimento , Astrágalo (Planta)/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Humanos , Metabolômica , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Fitoterapia , Cogumelos Shiitake/química , Cogumelos Shiitake/crescimento & desenvolvimento , Cogumelos Shiitake/metabolismo , Microambiente Tumoral
3.
J Sci Food Agric ; 101(13): 5618-5626, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33709431

RESUMO

BACKGROUND: Metabolite formation is a physiological stress response during the growth and development of shiitake mushrooms (Lentinula edodes). The characteristic flavor metabolites are important quality components in shiitake mushrooms. To investigate the formation mechanisms of characteristic flavor metabolites, transcriptome analyses were performed on shiitake mushrooms harvested at different growth stages. RESULTS: In total, 30 genes related to the synthesis of characteristic volatiles of mushrooms were identified via screening. Through KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis of the selected genes and correlation analyses of gene expressions, the main volatile synthesis pathways were determined as histidine metabolism, glutathione metabolism and biosynthesis of unsaturated fatty acids. Gene cluster and correlation analyses were performed to clarify the combined effects of different genes in the enzymatic reactions. Further, a correlation network of candidate genes was built based on the gene expression levels. CONCLUSION: The activities of flavor synthases and the content of characteristic flavor metabolites were analyzed; the enzyme activity changes and metabolic product distribution sites were clarified. A synthesis and regulation network was constructed for the candidate genes and characteristic volatiles, and information was obtained for 16 hub genes. Moreover, it was essential to identify and characterize the key genes and synthases involved in the synthesis of the characteristic volatiles of mushrooms. This information provides us with a better understanding of the biosynthesis and regulation of the volatiles, which will lay the foundation for improving the quality of shiitake mushrooms. © 2021 Society of Chemical Industry.


Assuntos
Aromatizantes/metabolismo , Proteínas Fúngicas/genética , Cogumelos Shiitake/genética , Vias Biossintéticas , Aromatizantes/química , Proteínas Fúngicas/metabolismo , Redes Reguladoras de Genes , Odorantes/análise , Cogumelos Shiitake/química , Cogumelos Shiitake/metabolismo , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/metabolismo
4.
Food Chem ; 351: 129227, 2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-33647695

RESUMO

Cold chain transportation is an important link in postharvest logistics of agricultural products. In current study, we developed a novel water-based phase change coolant (PCC), which showed longer effectiveness in maintaining low temperature condition compared with ice, and applied in preserving the postharvest mushrooms. The results showed that the novel PCC effectively inhibited water loss, as well as maintained quality attributes including firmness, color, phenolics, flavonoids, and thus prolonged the shelf-life of mushrooms. Low temperature condition created by the novel PCC treatment maintained high level of energy charge by activating the activities of SDH, CCO, H+-ATPase and Ca2+-ATPase, resulting in the delay of postharvest senescence. In addition, sufficient energy supply decreased the consumption of glutamate as carbon skeleton by inhibiting GDH activity, improved glutamate accumulation, and therefore maintained sensory properties as a result. Thus, the novel PCC might be an excellent substitute for ice in cold chain transportation of mushrooms.


Assuntos
Temperatura Baixa , Metabolismo Energético/efeitos dos fármacos , Qualidade dos Alimentos , Ácido Glutâmico/metabolismo , Transição de Fase , Cogumelos Shiitake/química , Cogumelos Shiitake/metabolismo , Cor , Cogumelos Shiitake/efeitos dos fármacos , Água/química , Água/farmacologia
5.
Food Chem ; 351: 129230, 2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-33626468

RESUMO

Water holding capacity (WHC) is an important factor for the evaluation of rehydrated shiitake mushroom as WHC largely affects sensory attributes. Here, WHC of the rehydrated shiitake mushroom as affected by different hot-air drying temperature was investigated from aspects of cell membrane integrity, status of cell wall fibrous material and protein denaturation. Among the three factors, protein denaturation played the most important role to affect the WHC of rehydrated mushroom as protein denaturation was closely related to the maintenance of porous structure after rehydration. Effect of status of the cell wall fibrous material to the WHC changes was relatively weak and was largely correlated to the WHC changes at high temperature (>80 °C). Cell membrane was unlikely to contribute to WHC changes of the rehydrated shiitake mushroom due to the loss of integrity.


Assuntos
Membrana Celular/metabolismo , Parede Celular/metabolismo , Temperatura Alta , Proteínas de Plantas/metabolismo , Cogumelos Shiitake/citologia , Cogumelos Shiitake/metabolismo , Água/metabolismo , Proteínas de Plantas/química , Desnaturação Proteica
6.
Food Chem ; 347: 128998, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33453580

RESUMO

Eight carbon (C8) compounds are the key characteristic flavors of mushrooms. The quantitative analysis of the volatiles in mushrooms is challenging especially with the unattended HS-SPME-GC-MS. An unattended HS-SPME-GC-MS/MS in combination with novel sample preparation of the complete control of volatile emissions was developed for the quantitation of the C8 volatiles in mushrooms. The sample preparation strategy was composed of freeze-drying, rehydration, and the addition of a 15% citric acid solution. With this strategy, the volatile emission from mushroom was fully controlled at a certain time point. This method was found to be highly reliable, sensitive, precise, and accurate. This method was successfully applied to measure the contents of the C8 volatiles in the beech, button, and shiitake mushrooms. 1-Octene-3-ol was the most predominant compound in the mushrooms, representing 62.4, 69.0, and 89.2% of the total C8 volatiles in the beech, button, and shiitake mushrooms, respectively.


Assuntos
Agaricales/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Compostos Orgânicos Voláteis/análise , Agaricales/metabolismo , Aromatizantes/análise , Limite de Detecção , Análise de Componente Principal , Cogumelos Shiitake/química , Cogumelos Shiitake/metabolismo , Microextração em Fase Sólida , Espectrometria de Massas em Tandem , Compostos Orgânicos Voláteis/isolamento & purificação
7.
J Antibiot (Tokyo) ; 73(10): 687-696, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32733077

RESUMO

Mushrooms have been attracting attention as a source of bioactive compounds for the development of dietary supplements and medicines. Many researchers have reported pharmacological effects of edible mushrooms, and have isolated and identified bioactive substances. Lentinula edodes (shiitake) and Flammulina velutipes (enokitake) are the cultivated edible mushrooms that are popular throughout the world. In L. edodes, polyacetylenes and sulfur compounds have been shown to display antimicrobial activity. In F. velutipes, many types of bioactive terpenes have been reported from mycelium culture filtrate or solid culture substrate. This article reviews the bioactive metabolites of low-molecular weight from L. edodes and F. velutipes.


Assuntos
Anti-Infecciosos/isolamento & purificação , Flammulina/química , Cogumelos Shiitake/química , Flammulina/metabolismo , Polímero Poliacetilênico/isolamento & purificação , Metabolismo Secundário , Cogumelos Shiitake/metabolismo , Relação Estrutura-Atividade , Terpenos/isolamento & purificação
8.
Food Chem ; 330: 127214, 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-32521398

RESUMO

A high glycaemic index diet causes a rapid increase in blood sugar level and may lead to chronic metabolic disorders such as obesity and type 2 diabetes. Shiitake is rich in bioactive compounds. Wheat flour noodles were enriched with shiitake (Lentinus edodes) powder (cap, stem, whole) at different levels to investigate the effects of shiitake addition on the nutritional composition, physical and textural properties. In vitro digestion was conducted to determine the glycaemic glucose equivalents and bioaccessibility of antioxidants in digesta. The addition of 15% shiitake stem powder in the noodles resulted in a significant (p < 0.05) decrease in reducing sugars released after in vitro digestion. Digesta also exhibited cellular antioxidant ability on IEC-6 cells after H2O2-induced oxidative stress. These results show the potential beneficial use of shiitake, especially the stem, as a high-value ingredient to improve the nutritional profile and reduce the glycaemic index of foods.


Assuntos
Antioxidantes/metabolismo , Cogumelos Shiitake/metabolismo , Triticum/metabolismo , Linhagem Celular , Digestão , Farinha , Mucosa Gástrica , Índice Glicêmico , Humanos , Estômago
9.
PLoS One ; 15(3): e0230680, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32218597

RESUMO

Lentinula edodes is one of the most popular edible mushrooms worldwide and contains important medicinal components such as lentinan, ergosterol, and eritadenine. Mushroom metabolism is regulated by the mycelia and fruit body using light; however, in mushrooms, the underlying molecular mechanisms controlling this process as well as light-induced gene expression remain unclear. Therefore, in this study, we compared morphological changes and gene expression in the fruit bodies of L. edodes cultivated under blue light and continuous darkness. Our results showed that blue light primarily induced pileus growth (diameter and thickness) compared to dark cultivation. Alternatively, stipe length development was promoted by dark cultivation. We also performed RNAseq on L. edodes under the blue light/dark cultivation conditions. A total of 12,051 genes were used for aligning the Illumina raw reads and 762 genes that showed fold change cut-offs of >|2| and significance p-values of <0.05 were selected under blue light condition. Among the genes which showed two-fold changed genes, 221 were upregulated and 541 were downregulated. In order to identify blue light induced candidate genes, differentially expressed genes (DEGs) were selected according to 4-fold changes and validated by RT-PCR. We identified 8 upregulated genes under blue light condition, such as DDR48-heat shock protein, Fasciclin-domain-containing protein and carbohydrate esterase family 4 protein, FAD NAD-binding domain-containing protein that are involved in morphological development of primordium and embryonic muscle development, cell adhesion and affect the structure of cellulosic and non-cellulosic cell walls of fruit body development, and photoreceptor of blue light signaling for fruit body and pigment development, respectively. This study provides valuable insights into the molecular mechanisms underlying the role of blue light in mushroom growth and development and can thus contribute to breeding programs to improve mushroom cultivation.


Assuntos
Regulação para Baixo/efeitos da radiação , Luz , Cogumelos Shiitake/genética , Regulação para Cima/efeitos da radiação , Proteínas Fúngicas/genética , RNA Fúngico/química , RNA Fúngico/isolamento & purificação , RNA Fúngico/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de RNA , Cogumelos Shiitake/metabolismo
10.
Fungal Biol ; 124(2): 135-143, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32008754

RESUMO

Lentinula edodes is a widely-produced mushroom in China that forms a brown film via pigment accumulation on mature mycelial surfaces to ensure high-quantity and high-quality fruiting body formation. Here, ultraviolet-visible, infrared spectra, and elemental analyses predicted that the pigment in the brown film was melanin. Electron microscopy revealed the size, morphological characteristics, accumulation, and morphogenesis of electron-dense material, which were similar to those of melanin, as well as subcellular structural changes during brown film formation. The electron-dense material appeared as granules, vesicles, and polymers. The accumulation of electron-dense materials on the cell wall was followed plasmolysis, plasma membrane disruption, electron-dense material accumulation in the interstitial space, and gradual accumulation on the outer cell wall. Dolipore septa degradation and morphogenetic cell death occurred during browning. In the final stage of browning, the dolipore septum disappeared and the cell was nearly empty. This study provides a cytological foundation for evaluating the regulation of brown film formation in L. edodes.


Assuntos
Melaninas/metabolismo , Cogumelos Shiitake , Carpóforos/metabolismo , Proteínas Fúngicas , Microscopia Eletrônica de Transmissão , Micélio/metabolismo , Micélio/ultraestrutura , Pigmentos Biológicos , Cogumelos Shiitake/crescimento & desenvolvimento , Cogumelos Shiitake/metabolismo , Análise Espectral
11.
Carbohydr Polym ; 229: 115521, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31826486

RESUMO

A polysaccharide-enriched extract obtained from Lentinula edodes was submitted to several purification steps to separate three different D-glucans with ß-(1→6), ß-(1→3),(1→6) and α-(1→3) linkages, being characterized through GC-MS, FT-IR, NMR, SEC and colorimetric/fluorimetric determinations. Moreover, in vitro hypocholesterolemic, antitumoral, anti-inflammatory and antioxidant activities were also tested. Isolated glucans exerted HMGCR inhibitory activity, but only ß-(1→6) and ß-(1→3),(1→6) fractions showed DPPH scavenging capacity. Glucans were also able to lower IL-1ß and IL-6 secretion by LPS-activated THP-1/M cells and showed cytotoxic effect on a breast cancer cell line that was not observed on normal breast cells. These in vitro results pointed important directions for further in vivo studies, showing different effects of each chemical structure of the isolated glucans from shiitake mushrooms.


Assuntos
Glucanos/química , Cogumelos Shiitake/metabolismo , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Antioxidantes/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Glucanos/isolamento & purificação , Glucanos/metabolismo , Humanos , Hidroximetilglutaril-CoA Redutases/química , Hidroximetilglutaril-CoA Redutases/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos/toxicidade , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo
12.
Food Chem ; 309: 125738, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31706679

RESUMO

In the present study, the impact of ultraviolet (UV)-C treatment and ultrafine grinding on the conversion of ergosterol to vitamin D2, physiochemical properties, and antioxidant properties of shiitake and Jew's ear was assessed. After exposure to UV-C, vitamin D2 contents of both the mushroom samples has increased significantly (p < 0.05). Whereas, ultrafine grinding along with UV-C treatment has a synergistic effect on bioconversion of ergosterol to vitamin D2 and this effect is more prominent in low dose UV-C irradiation groups (2 kJ/m2). Ultrafine grinding significantly (p < 0.05) improved the water holding capacity (WHC), water solubility index (WSI) and polysaccharide dissolution rate (PDR). However, UV-C treatment led to insignificant changes in the physiochemical properties of mushroom samples. A significant improvement was also observed in the antioxidant profiles especially tannin contents of mushrooms followed by the ultrafine grinding and UV-C treatment.


Assuntos
Agaricales/metabolismo , Agaricales/efeitos da radiação , Antioxidantes/metabolismo , Ergocalciferóis/metabolismo , Ergosterol/metabolismo , Cogumelos Shiitake/metabolismo , Cogumelos Shiitake/efeitos da radiação , Agaricales/química , Antioxidantes/química , Biotransformação , Ergocalciferóis/química , Ergosterol/química , Cogumelos Shiitake/química , Raios Ultravioleta
13.
J Biosci Bioeng ; 129(4): 514-518, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31708427

RESUMO

Okara, despite being a soybean processing by-product, still holds many nutrients. Thus, considerable attention has been recently paid to its reuse. In this study, solid-state fermentation was performed using Ganoderma lucidum and Lentinus edodes. Antioxidant activity and bioactive compound levels in G. lucidum-fermented okara (GLFO) and L. edodes-fermented okara (LEFO) were assayed. Antiosteoporosis bioactivity was evaluated using an animal model. The results demonstrated that solid-state fermentation significantly improved the antioxidant activity and bioactive compound levels. Furthermore, GLFO and LEFO increased trabecular bone volume, although only the GLFO-treated group exhibited significantly improved trabecular separation compared with the bilateral ovariectomy-treated control group. GLFO-related outcomes were superior to those of LEFO. The results demonstrate that okara products are effective for treating postmenopausal osteoporosis in humans.


Assuntos
Conservadores da Densidade Óssea , Fermentação , Extratos Vegetais , Reishi/química , Cogumelos Shiitake/química , Soja/química , Animais , Biomassa , Reatores Biológicos , Conservadores da Densidade Óssea/isolamento & purificação , Conservadores da Densidade Óssea/metabolismo , Conservadores da Densidade Óssea/farmacologia , Conservadores da Densidade Óssea/uso terapêutico , Suplementos Nutricionais , Avaliação Pré-Clínica de Medicamentos , Feminino , Camundongos , Camundongos Endogâmicos ICR , Osteoporose/tratamento farmacológico , Ovariectomia , Extratos Vegetais/química , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Reishi/metabolismo , Cogumelos Shiitake/metabolismo , Soja/metabolismo
14.
Int J Med Mushrooms ; 22(8): 781-791, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33389872

RESUMO

The shiitake culinary-medicinal mushroom Lentinus edodes (=Lentinula edodes) has been used as a nutritional complement. This study evaluated, through reprotox tests, the consumption of L. edodes in pregnant rats, considering it as a functional food. Pregnant rats were daily exposed to L. edodes before implantation (LeB)-from 1 to 19 days of gestation, and after implantation (LeA)-from 9 to 19 days of gestation, compared with controls. On the 20th day of gestation, cesarean sections were performed. Blood was collected and hematological parameters (hemoglobin, hematocrit, white and red blood cells and platelets) were analyzed. Moreover, albumin, calcium, creatine kinase, alkaline phosphatase, transferases, creatinine, urea, triglycerides, cholesterol, lipase, glucose, and insulin were assessed in serum. Organs were collected and weighed, and the fetuses were analyzed morphologically by body measurements. The consumption of L. edodes reduced triglycerides levels and there were no changes in maternal weight, biochemical and hematological parameters, organ weight, and reproductive capacity. There were no morphological changes in the fetuses' body measurements, suggesting possible safety in ingestion of mushroom. Reprotox tests used to evaluate the daily consumption of L. edodes points out potentiality as a functional food, suggesting safety during the gestational period.


Assuntos
Desenvolvimento Fetal , Gravidez/metabolismo , Cogumelos Shiitake/metabolismo , Animais , Colesterol/metabolismo , Feminino , Feto/metabolismo , Alimento Funcional/análise , Humanos , Masculino , Fenômenos Fisiológicos da Nutrição Materna , Ratos , Ratos Wistar , Triglicerídeos/metabolismo
15.
Food Res Int ; 125: 108517, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31554126

RESUMO

Volatile compounds of Lentinula edodes grown in different culture substrate (CS) formulations were analyzed to reveal (i) the relationship between volatile compound production and CS formulations, (ii) the contribution of volatile compounds to L. edodes flavor, (iii) the activities of LOX and γ-GGT enzymes, (iv) γ-GGT gene expression, and (v) the correlation between enzyme activity and volatile compound production. Our results showed that 82 kinds of volatile compounds were analyzed; 25 volatile compounds were considered key flavor components, and sulfur containing compounds, eight-carbon compounds, and aldehyde compounds also had great contributions to mushroom flavor. Bagasse could be used as a partial substitute for sawdust as a carbon source. LOX and γ-GGT activities showed a weak correlation with the volatile end products. The results indicated that the mechanisms by which CS formulations influence volatile compounds production were complex.


Assuntos
Agricultura/métodos , Cogumelos Shiitake , Compostos Orgânicos Voláteis/análise , Cogumelos Shiitake/química , Cogumelos Shiitake/genética , Cogumelos Shiitake/metabolismo
16.
J Agric Food Chem ; 67(41): 11373-11379, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31539240

RESUMO

Cadmium bioremediation with metal-binding proteins is primarily conducted using metallothioneins (MTs). However, in the present study, we investigated a non-MT cadmium-binding protein from Lentinula edodes (LECBP) as a remediation tool for cadmium biosorption in Escherichia coli. The results indicated that the expression of LECBP significantly enhanced the cadmium biosorption capacity of transgenic E. coli. The secondary structure and conformation of LECBP were changed after binding with cadmium as evidenced by circular dichroism and fluorescence spectroscopy. The results of Fourier transform infrared spectroscopy indicated that carboxyl oxygen and amino nitrogen atoms were involved in the interaction between LECBP and cadmium. The results further demonstrated that glutamic acid and histidine residues are the potential binding sites. Our results have thus provided new insights into cadmium bioremediation in an aquatic environment.


Assuntos
Cádmio/metabolismo , Metalotioneína/metabolismo , Proteínas de Plantas/metabolismo , Cogumelos Shiitake/genética , Sítios de Ligação , Biodegradação Ambiental , Cádmio/química , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Metalotioneína/química , Metalotioneína/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Cogumelos Shiitake/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
17.
J Biotechnol ; 302: 42-47, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31247237

RESUMO

Vitreoscilla hemoglobin (VHb), encoded by the Vitreoscilla hemoglobin gene (vgb), is highly effective at binding oxygen and delivering it to both prokaryotes and eukaryotes under hypoxic conditions. In this study, we introduced the vgb gene into shiitake mushrooms, and the mycelia of the transformatants grew faster. In particular, they spread into the solid substrate located in the lower part of the test tubes and bags where the oxygen was hypoxic and produced more ß-glucan and plant biomass degrading enzymes compared to the original strain. The maximum growth rate of the transformants was 8.5%-15.9% higher than that of the original strain on sawdust-based cultures in plastic bags. The laccase and amylase activities were 17.7%-40.3% and 16.7%-37.9% higher than that of the original strain, respectively. In addition, the ß-glucan contents of the transformant mycelia from the submerged fermentation were 12.9%-24.0% higher than that of the original strain. These results reveal that the expression of VHb in mushroom fungi promots the mycelial growth in solid-state cultures under the hypoxic condition as well as enhances ß-glucan and plant biomass degrading enzymes production.


Assuntos
Biomassa , Cogumelos Shiitake/metabolismo , Vitreoscilla/metabolismo , Amilases/metabolismo , Fermentação/fisiologia , Lacase/metabolismo
18.
Int J Mol Sci ; 20(9)2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31083449

RESUMO

To clarify the physiological mechanism of the Lentinula edodes (L. edodes) response to high-temperature stress, two strains of L. edodes with different tolerances were tested at different durations of high temperature, and the results showed that there were significant changes in their phenotypes and physiology. To further explore the response mechanism, we established a targeted GC-MS-based metabolomics workflow comprising a standardized experimental setup for growth, treatment and sampling of L. edodes mycelia, and subsequent GC-MS analysis followed by data processing and evaluation of quality control (QC) measures using tailored statistical and bioinformatic tools. This study identified changes in the L. edodes mycelial metabolome following different time treatments at high temperature based on nontargeted metabolites with GC-MS and further adopted targeted metabolomics to verify the results of the analysis. After multiple statistical analyses were carried out using SIMCA software, 74 and 108 differential metabolites were obtained, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the metabolic pathways with significant changes included those related to the following: amino acid metabolism, the glycolysis pathway, the tricarboxylic acid (TCA) cycle, and sugar metabolism. Most amino acids and carbohydrates enriched in these metabolic pathways were upregulated in strain 18, downregulated in strain 18N44, or the synthesis in strain 18 was higher than that in strain 18N44. This result was consistent with the physiological phenotypic characteristics of the two strains under high-temperature stress and revealed the reason why strain 18N44 was more heat-sensitive. At the same time, under high temperature, the decrease of intermediate products in glycolysis and the TCA cycle resulted in carbon starvation and insufficient energy metabolism, thus inhibiting the growth of L. edodes. In addition, the results also showed that the metabolites produced by different L. edodes strains under high-temperature stress were basically the same. However, different strains had species specificity, so the changes in the content of metabolites involved in the response to high-temperature stress were different. This provides a theoretical basis for further understanding the mechanism of the L. edodes response to high temperature and can be used to establish an evaluation system of high-temperature-resistant strains and lay the foundation for molecular breeding of new L. edodes strains resistant to high temperature.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Temperatura Alta , Metaboloma , Metabolômica , Micélio/metabolismo , Cogumelos Shiitake/metabolismo , Estresse Fisiológico , Micélio/crescimento & desenvolvimento , Reprodutibilidade dos Testes , Cogumelos Shiitake/crescimento & desenvolvimento
19.
J Agric Food Chem ; 67(4): 1261-1268, 2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30623660

RESUMO

Many organisms possess the ability to produce metal-binding proteins to absorb cadmium. Metallothioneins, an important family of cysteine-rich metal-binding proteins, have been isolated and well characterized. However, Lentinula edodes may have a different type of cadmium-binding protein that contains fewer cysteine residues. In the present study, we purified a cadmium-binding protein from L. edodes (LECBP) by gel filtration and anion exchange chromatography and then identified LECBP by LC-MS/MS. We found LECBP to be a novel cadmium-binding protein, which contained 220 amino acid residues but no cysteine residue. LECBP had a high binding affinity for Cd(II) with a Kd value of 97.3 µM. The percentages of α-helix, ß-sheet, ß-turn, and random coil in LECBP were 15.7%, 39.4%, 8.0%, and 37.1%, respectively. In addition, high temperatures and an acidic environment influenced the conformation of LECBP. Our results will thus provide a new perspective to understand the mechanism of cadmium accumulation in L. edodes.


Assuntos
Proteínas Fúngicas/química , Proteínas Fúngicas/isolamento & purificação , Metalotioneína/química , Metalotioneína/isolamento & purificação , Cogumelos Shiitake/química , Sequência de Aminoácidos , Cádmio/química , Cádmio/metabolismo , Cromatografia em Gel , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Cinética , Espectrometria de Massas , Metalotioneína/genética , Metalotioneína/metabolismo , Dados de Sequência Molecular , Conformação Proteica em alfa-Hélice , Cogumelos Shiitake/genética , Cogumelos Shiitake/metabolismo
20.
Int J Med Mushrooms ; 21(9): 909-919, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32450029

RESUMO

The DnaJ proteins, also called heat-shock protein 40 based on their molecular weight, play significant roles in organism growth and development and resistance to abiotic and biotic stresses. However, studies on the DnaJ gene family in Lentinus edodes (= Lentinula edodes) are less well known. In this study, 29 putative L. edode DnaJ genes (LeDnaJ01 to LeDnaJ29) were identified using bioinformatics analysis and were classified into four groups according to the presence of the J protein and zinc finger as well as C-terminal domain. Multiple cis elements related to the phytohormone and stresses were found in the promotor region of the LeDnaJ genes. In addition, qRT-PCR analysis revealed that 79.31% of LeDnaJ genes were induced by cadmium, 55.17% were induced by Trichoderma atroviride, and 37.93% were induced by heat stress, indicating that the LeDnaJ proteins might participate in the response of L. edodes to the multiple stresses. Meanwhile, qRT-PCR analysis also revealed that all LeDnaJs are expressed in at least one development stage, indicating that they could be involved in the process of L. edodes growth and development and the response to the abiotic and biotic stresses. Taken together, these results advance the functional analysis of DnaJ genes in Basidiomycetes.


Assuntos
Proteínas Fúngicas/genética , Proteínas de Choque Térmico HSP40/genética , Cogumelos Shiitake/genética , Cogumelos Shiitake/metabolismo , Transcriptoma , Cádmio/farmacologia , Biologia Computacional , Temperatura Alta , Regiões Promotoras Genéticas , Estresse Fisiológico
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