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1.
Biosci. j. (Online) ; 36(6): 2238-2246, 01-11-2020. graf
Artigo em Inglês | LILACS | ID: biblio-1148295

RESUMO

Lentinus crinitus is a medicinal basidiomycete, little studied regarding the basic cultivation conditions, which is used in bioremediation and consumed by native Indians from the Brazilian Amazon. Also, it produces a fungal secondary metabolite panepoxydone that has been described as an essential regulator of the inflammatory and immune response. This study aimed to evaluate basic conditions of temperature, pH, and nitrogen concentration and source in the cultivation of L. crinitus mycelial biomass. In order to evaluate fungal growth temperature, 2% malt extract agar (MEA) medium, pH 5.5, was utilized from 19 to 40 °C. For pH, MEA had pH adjusted from 2 to 11 and cultivated at 28 °C. Urea or soybean meal was added to MEA to obtain final concentration from 0.5 and 16 g/L of nitrogen, pH of 5.5, cultivated at 28 °C. The best temperature growth varies from 31 to 34 ºC and the optimal one is 32.7º C, and the best pH ranges from 4.5 to 6.5 and the optimal one is 6.1. Protein or non-protein nitrogen concentration is inversely proportional to the mycelial biomass growth. Nitrogen concentrations of 2.0 g/L soybean meal and urea inhibit mycelial biomass growth in 11% and 12%, respectively, but high concentrations of 16.0 g/L nitrogen inhibit the growth in 46% and 95%, respectively. The fungus is robust and grows under extreme conditions of temperature and pH, but smaller adaptation with increasing nitrogen concentrations in the cultivation medium, mainly non-protein nitrogen.


Lentinus crinitus é um basidiomiceto medicinal consumido por índios nativos da Amazônia brasileira. Este fungo tem sido estudado quanto ao potencial de biorremediação de metais, mas ainda carece de estudos sobre às condições básicas de crescimento. L. crinitus produz panepoxidona - um metabólito secundário fúngico - descrito como regulador da resposta inflamatória e imune em células animais. Este trabalho teve como objetivo avaliar as condições básicas de temperatura, pH e concentração e fonte de nitrogênio para o crescimento micelial de L. crinitus. O fungo foi crescido em meio agar extrato de malte a 2% (MEA), pH 5,5 e mantido entre 19 e 40 °C. Para a avaliação de pH o MEA teve o pH ajustado de 2 a 11 e o crescimento foi realizado a 28 °C. As fontes de nitrogênio estudadas foram a uréia e o farelo de soja adicionado ao MEA para obter entre 0,5 a 16 g/L de nitrogênio, pH de 5,5, cultivado a 28 ° C. A melhor faixa temperatura para o crescimento micelial foi de 31 a 34 ºC com ótimo a 32,7 º C; a melhor faixa de pH de 4,5 a 6,5 e com ótimo de 6,1. A concentração de nitrogênio proteico ou não proteico é inversamente proporcional ao crescimento do fungo. Concentrações de nitrogênio de 2,0 g/L reduzem o crescimento da biomassa micelial em 11% e 12%, respectivamente e meios com nitrogênio de 16,0 g/L reduzem o crescimento em 46% e 95%, respectivamente. O fungo é robusto e cresce sob condições extremas de temperatura e pH, mas menor adaptação em meios com alta concentração de nitrogênio, principalmente não proteico.


Assuntos
Biomassa , Lentinula , Cultura Axênica , Micelas , Ureia
2.
Biosci. j. (Online) ; 36(3): 924-931, 01-05-2020. tab
Artigo em Inglês | LILACS | ID: biblio-1146988

RESUMO

Fungi are capable of sensing light from ultraviolet to far-red and they use light as a source of information about the environment anticipating stress and adverse conditions. Lentinus crinitus is a lignin-degrading fungus which produces laccase and other enzymes of biotechnological interest. The effect of blue light on fungal enzymatic activity has been studied; however, it has not been found studies on the effect of the blue light on carbohydrate-active enzymes and on mycelial biomass production of L. crinitus. We aimed to investigate carbohydrate-active enzymes activity and mycelial biomass production of L. crinitus cultivated under continuous illumination with blue light. L. crinitus was cultivated in malt extract medium in the dark, without agitation, and under continuous illumination with blue light-emitting diodes. The blue light reduced the total cellulase, pectinase and xylanase activities but increased the endoglucanase activity. Blue light reduced the mycelial growth of L. crinitus in 26% and the enzymatic activity-to-mycelial biomass ratio (U mg-1 dry basis) increased in 10% total cellulase, 33% endoglucanase, and 16% pectinase activities. Also, it is suggested that L. crinitus has a photosensory system and it could lead to new process of obtaining enzymes of biotechnological interest.


Fungos são capazes de sentir a luz com comprimentos de onda que variam do ultravioleta ao infravermelho e usam a luz como fonte de informação sobre o ambiente, antecipando condições adversas e de estresse. Lentinus crinitus é um fungo ligninolítico que produz lacase e outras enzimas de interesse biotecnológico. O efeito da luz azul na atividade enzimática de fungos já foi estudado, contudo, ainda não há estudos sobre o efeito da luz azul na produção de enzimas ativas sobre carboidratos (CAZymes, carbohydrate-active enzymes) e de biomassa micelial de L. crinitus. O objetivo deste estudo foi investigar a avitivade de CAZymes e a produção de biomassa micelial de L. crinitus cultivado sob iluminação continua com luz azul. L. crinitus foi cultivado em meio extrato de malte, sem agitação, na ausência de luz e sob luz continua fornecida por diodos emissores de luz azul. A luz azul reduziu a atividade de cellulase total, pectinase e xilanase, mas aumentou a atividade de endoglucanase. A luz azul reduziu o crescimento micelial de L. crinitus em 26% e aumentou a razão atividade enzimática/biomassa micelial (U mg-1 em base seca) de cellulase total em 10%, endoglucanase em 33% e pectinase em 16%. Além disso, sugere-se que L. crinitus possua um sistema fotossensorial que poderia ser explorado para a otimização de bioprocessos que visam a obtenção de enzimas de interesse biotecnológico.


Assuntos
Poligalacturonase , Lentinula , Celulases , Luz
3.
Electron. j. biotechnol ; 32: 6-12, Mar. 2018. tab, graf, ilus
Artigo em Inglês | LILACS | ID: biblio-1022493

RESUMO

Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier.


Assuntos
Basidiomycota/metabolismo , Proteínas Fúngicas/genética , Lentinula/genética , Lentinula/metabolismo , Transformação Genética , Basidiomycota/enzimologia , Leveduras , Proteínas Fúngicas/metabolismo , Southern Blotting , Clonagem Molecular , Agrobacterium tumefaciens/metabolismo , Análise de Sequência , Emulsificantes , Eletroforese em Gel de Poliacrilamida , Reação em Cadeia da Polimerase em Tempo Real , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Microscopia de Fluorescência
4.
Vitae (Medellín) ; 23(2): 109-118, 2016. Ilustraciones
Artigo em Inglês | LILACS, COLNAL | ID: biblio-988465

RESUMO

Background: Natural compounds are a good source for the development of antiretroviral drugs with low cytotoxicity. The laccase enzyme, produced by fungi of the genera Ganoderma sp. and Lentinus sp., inhibits the reverse transcriptase (RT) of the human immunodeficiency virus 1 (HIV-1), in cell-free models in vitro. Objetives: In this study we evaluated the anti-HIV-1 activity of the enzymatic extracts (EE) enriched with laccase, produced by two native species of fungi of the same genera in an in vitro cell culture model. Methods: The inhibition of viral replication was performed using the U373-MAGI cell line infected with recombinant viruses in the presence/absence of the EE and 48 hpi, the percentage of infected cells was evaluated by flow cytometry for green fluorescent protein ­GFP­ and ELISA for p24. The inhibition of the RT was determined by quantification of early and late products of reverse transcription using quantitative PCR. Results: The EEs from Ganoderma sp. and Lentinus sp. inhibited the replication of HIV-1 between 80 and 90% and decreased the production of early and late transcripts between 55,5%-91,3% and 82,1%-93,6% respectively. The EE from Lentinus sp. had the best selectivity index (SI: 8.3). Conclusions: These results suggest the potential anti-HIV-1 activity of the EE for the exploration of an alternative therapy against HIV-1 infection.


Antecedentes: Los compuestos naturales son una buena fuente para el desarrollo de fármacos antirretrovirales con baja citotoxicidad. La enzima lacasa, producida por hongos del género Ganoderma sp. y Lentinus sp., inhibe la transcriptasa reversa (TR) del virus de la inmunodeficiencia humana tipo 1 (VIH-1), en modelos in vitro, libres de células. Objetivos: En este estudio se evaluó la actividad anti-VIH-1 del extracto enzimático (EE) enriquecido con lacasa, producida por dos especies nativas de hongos de los mismos géneros en un modelo in vitro de cultivo celular. Métodos: La inhibición de la replicación viral se realizó usando la línea celular U373-MAGI infectada con virus recombinantes en la presencia/ ausencia del EE y 48 hpi, el porcentaje de células infectadas se evaluó mediante citometría de flujo para GFP y ELISA para p24. La inhibición de la TR se determinó mediante la cuantificación de los productos tempranos y tardíos de la transcripción reversa utilizando una PCR cuantitativa. Resultados: El EE de Ganoderma sp. y Lentinus sp. inhibió la replicación del VIH-1 entre el 80 y 90% y disminuyó la producción de transcriptos tempranos y tardíos entre el 55,5% -91,3% y 82,1% -93,6%, respectivamente. El EE de Lentinus sp. mostró el mejor índice de selectividad (IS: 8.3). Conclusiones: Estos resultados sugieren el potencial anti-VIH-1 del EE para la exploración de una terapia alternativa contra la infección por el VIH-1.


Assuntos
Humanos , Antivirais , HIV-1 , Produtos Biológicos , Lentinula , Ganoderma , Lacase
5.
Braz. j. med. biol. res ; 43(11): 1054-1061, Nov. 2010. ilus, tab
Artigo em Inglês | LILACS | ID: lil-564126

RESUMO

Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity onTrypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 μg/mL (40.3 μM and 47.6 μM for hypnophilin and panepoxydone, respectively; ~100 percent), hypnophilin has a slightly greater inhibitory activity (~71 percent) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70 percent) compared to panepoxydone (69 percent AMA inhibition and 91 percent PBMC inhibition). Hypnophilin and panepoxydone at 1.25 μg/mL had 67 percent inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16+ and CD14+ cells and down-regulated CD19+, CD4+ and CD8+ cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.


Assuntos
Humanos , Antiprotozoários/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Leishmania/efeitos dos fármacos , Lentinula/química , Extratos Vegetais/farmacologia , Sesquiterpenos/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Antígenos CD/efeitos dos fármacos , Compostos Bicíclicos Heterocíclicos com Pontes/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Desenho de Fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Sesquiterpenos/isolamento & purificação
6.
Mem. Inst. Oswaldo Cruz ; 103(3): 263-270, May 2008. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-485218

RESUMO

The fungus Lentinus strigosus (Pegler 1983) (Polyporaceae, basidiomycete) was selected in a screen for inhibitory activity on Trypanosoma cruzi trypanothione reductase (TR). The crude extract of L. strigosus was able to completely inhibit TR at 20 µg/ml. Two triquinane sesquiterpenoids (dihydrohypnophilin and hypnophilin), in addition to two panepoxydol derivatives (neopanepoxydol and panepoxydone), were isolated using a bioassay-guided fractionation protocol. Hypnophilin and panepoxydone displayed IC50 values of 0.8 and 38.9 µM in the TR assay, respectively, while the other two compounds were inactive. The activity of hypnophilin was confirmed in a secondary assay with the intracellular amastigote forms of T. cruzi, in which it presented an IC50 value of 2.5 µ M. Quantitative flow cytometry experiments demonstrated that hypnophilin at 4 µM also reduced the proliferation of human peripheral blood monocluear cells (PBMC) stimulated with phytohemaglutinin, without any apparent interference on the viability of lymphocytes and monocytes. As the host immune response plays a pivotal role in the adverse events triggered by antigen release during treatment with trypanocidal drugs, the ability of hypnophilin to kill the intracellular forms of T. cruzi while modulating human PBMC proliferation suggests that this terpenoid may be a promising prototype for the development of new chemotherapeutical agents for Chagas disease.


Assuntos
Animais , Bovinos , Humanos , Camundongos , Proliferação de Células/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Lentinula/química , NADH NADPH Oxirredutases/antagonistas & inibidores , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Inibidores Enzimáticos/isolamento & purificação , Citometria de Fluxo , Linfócitos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Tripanossomicidas/isolamento & purificação , Trypanosoma cruzi/enzimologia
7.
Vitae (Medellín) ; 12(2): 13-20, mar. 2005-sept. 2005. ilus, tab, graf
Artigo em Espanhol | LILACS | ID: lil-421259

RESUMO

En este articulo se presenta la producción de metabolitos secundarios de interés farmacéutico y alimentario obtenidos de los residuos del cultivo de plátano al emplearse como sustrato en un proceso de fermentación en estado sólido con el hongo de la podredumbre de la madera Lentinus crinitus.La producción de metabolitos se determina durante el crecimiento del microorganismo por un periodo de 21 días, en un sistema de fermentación conformado por 7 combinaciones de sustrato: tallo-fruto, hojas-fruto, hojas-tallo, hojas, tallo, fruto y hojas-tallo-fruto. La mejor combinación para la producción de las enzimas ligninoperoxidasa (LiP) y manganesoperoxidasa (MnP) es la conformada por hojas-tallo de plátano Musa paradisiaca. Esto es importante para aprovechar el enorme potencial que tienen estos residuos para la producción de enzimas peroxidasas, las cuales tienen muchas aplicaciones. También es importante destacar que anteriormente no se tienen reportes en la literatura a cerca de la producción de la enzima ligninoperoxidasa por el hongo Lentinus crinitus. En los sistemas donde se obtiene actividad enzimática: (tallo-fruto, hojas-fruto, hojas-tallo y hojas-tallo-fruto) se evalúa la presencia de los compuestos aromáticos ácido ferúlico, vainilla, ácido vainillínico y eugenol en los días 11, 16 y 21. Los análisis se hacen por cromatografía líquida de alta eficienia (HPLC). En los sistemas de mayor actividad enzimática conformados por la mezcla de hojas-tallos, los productos aromáticos de interés se detectaron en muy baja concentración. En el sistema conformado por hojas-frutos donde solo se destaca la producción de la enzima LiP, se encuentra ácido ferúlico, vainilla, ácido vainillínico y eugenol en cantidades importantes durante el día 16


Assuntos
Eugenol , Lentinula/isolamento & purificação , Lentinula/metabolismo , Resíduos Sólidos , Uso de Resíduos Sólidos , Zingiberales
8.
Braz. j. microbiol ; 32(3): 170-175, July-Sept., 2001. ilus, tab, graf
Artigo em Inglês | LILACS | ID: lil-316964

RESUMO

The growth of thirty-four Lentinula edodes strains submitted to different mycelial cultivation conditions (pH and temperature) was evaluated and strain variability was assessed by RAPD molecular markers. The growth at three pH values (5, 6 and 7) and four different temperatures (16, 25, 28 and 37§C) was measured using the in vitro mycelial development rate and water retention as parameters. Mycelial cultivation was successful at all pH tested, while the ideal temperature for mycelial cultivation ranged between 25 and 28§C. The water content was lower in strains grown at 37§C. Among 20 OPA primers (Operon Technologies, Inc.) used for the RAPD analyses, seventeen presented good polymorphism (OPA01 to OPA05, OPA07 to OPA14, OPA17 to OPA20). The clustering based on similarity coefficients allowed the separation of strain in two groups with different geographic origins.


Assuntos
Fungos , Técnicas In Vitro , Lentinula , Cultura , Linhagem
9.
Braz. j. microbiol ; 32(3): 176-178, July-Sept., 2001. tab, graf
Artigo em Inglês | LILACS | ID: lil-316965

RESUMO

Five species of edible mushrooms, Lentinula edodes, Pleurotus ostreatus, Pholiota nameko, Macrolepiota bonaerensis and Agaricus blazei, were tested for their potential to inhibit the in vitro growth of the pathogenic yeast Candida albicans. Only L. edodes had a fungistatic effect on this human pathogen. The inhibitory compound was produced intra and extracellularly in submersed L. edodes culture, and was also present in fresh and dehydrated mushroom basidiocarps. The fungistatic compound was heat sensitive and lost activity after 72 hours.


Assuntos
Candida albicans , Candidíase/diagnóstico , Candidíase/enzimologia , Fungos , Técnicas In Vitro , Lentinula , Cultura , Sintomas Patogenéticos
10.
Braz. j. microbiol ; 32(3): 206-210, July-Sept., 2001. tab, graf
Artigo em Inglês | LILACS | ID: lil-316970

RESUMO

The antibacterial activity of 35 isolates of Lentinula edodes, a shiitake mushroom, against Bacillus subtilis was evaluated by diffusion technique in agar with a semi-solid overlay. All isolates inhibited B. subtilis and the isolate Le1 promoted the formation of the largest inhibition zone. L. edodes Le1 also presented antibacterial activity against foodborne pathogens and food contaminant bacteria, particularly Gram-positive species. The antibacterial activity of the culture filtrate after 18-25 days of cultivation of L. edodes in broth at 25§C was high. The inhibitory activity was observed only in the organic layer when the culture filtrate was partitioned between ethyl acetate and water, suggesting that the inhibitory substances have low polarity. The silica gel thin-layer zone at Rf values of 0.63-0.80, developed in chloroform - acetone - ethyl acetate - methanol = 40:5:5:2, was responsible for the antibacterial activity against B. subtilis. The inhibitory activity of L. edodes was detectable in the culture filtrate after heat treatment at 100§C for 10 min and after storage at 4§C for 120 days.


Assuntos
Antígenos de Bactérias/imunologia , Antígenos de Bactérias/isolamento & purificação , Bactérias/patogenicidade , Técnicas In Vitro , Lentinula , Cromatografia em Camada Delgada , Meios de Cultura
11.
Braz. j. microbiol ; 32(1): 52-5, Jan.-Mar. 2001. tab
Artigo em Inglês | LILACS | ID: lil-297667

RESUMO

Four Lentinula edodes strains (Le10, 46, K2, Assai) were assessed for their antagonistic effect on four filamentous fungus species of agricultural importance (Helminthosporium euphorbiae, Helminthosporium sp, Fusarium solani and Phomopsis sojae) and on Alagoas serotype of Vesicular Stomatitis Virus (VSA). The L. edodes strains studied had variable effects on the filamentous fungi and on VSA. The K2 and Le10 strains were antagonistic on the fungi assessed and the 46 and K2 strains were efficient on the Vesicular Stomatitis Virus. The results widened the list of beneficial effects of L. edodes on the control and prevention of animal pathogenic virus and filamentous fungi.


Assuntos
Fungos , Lentinula , Vírus da Estomatite Vesicular Indiana
12.
Rev. microbiol ; 29(4): 286-8, out.-dez. 1998. graf
Artigo em Inglês | LILACS | ID: lil-251738

RESUMO

A modified method for direct determination of cellulolytic activity using Avicel colored with Remazol Brilliant Blue R (RBBR) in Agar test tubes in discussed. Refinaments were introduced in a simple method for qunatitation of cellulase activity, based on the release of dye from Avicel-RBBR medium by enzimatic hydrolysis. Modification in Avicel-dye preparation were enhanced and a spectrophotometer improved the precision of the collected data, since absorbance measurements could be done at the maxmum wavelenght for RBBR (595 nm).


Assuntos
Basidiomycota/metabolismo , Corantes , Técnicas In Vitro , Celulase/análise , Lentinula/metabolismo , Pleurotus/metabolismo , Espectrofotômetros
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