Mycelial biomass cultivation of Lentinus crinitus / Crescimento micelial de Lentinus crinitus

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.

Lentinus crinitus response to blue light on carbohydrate-active enzymes / Resposta de Lentinus crinitus à luz azul em enzimas ativas por carboidratos

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.

Evaluation of the cytotoxic effect of crude aqueous and ethanolic extracts isolated from Lentinus sp. on human cancer cell lines

Aims@#The aim of this study was to evaluate some chemical properties and the cytotoxic effect of aqueous and ethanolic crude polysaccharides extracted from five Lentinus sp. on human cancer cell lines. It was hypothesized that other species of the genus Lentinus could show the pharmacological actions as presence in Lentinus edodes, especially anticancer properties. @*Methodology and results@#Crude extracts of dried fruit bodies and mycelia from L. edodes, Lentinus sajor-caju, Lentinus swartzii, Lentinus squarrosulus and Lentinus velutinus were extracted using two solvents, hot water and 95% ethanol, and evaluated for their total carbohydrates, proteins, reducing sugar, phenol contents, and cytotoxicity. The yield of crude extracts was 33.6-205.3 mg/g dry weight of a sample. Cytotoxicity was determined with 10 mg/mL of crude aqueous and 1 mg/mL of crude ethanolic extracts by using the [3-(4,5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide] (MTT) method. All extracts showed non-cytotoxicity against the normal cell lines, LLC-MK2 and L929 cells. While, the extracts of L. edodes, L. sajor-caju, L. squarrosulus and L. velutinus displayed the cytotoxicity against the human cancer cell lines. @*Conclusion, significance and impact of study@#The crude aqueous and ethanolic extract from fruit bodies of L. velutinus and the ethanolic extract from fruit bodies of L. sajor-caju and L. squarrosulus displayed the adverse effect against the human cancer cell lines. Hence, these extracts are a potential source of bioactive compounds for cancer treatment.

Antioxidant activity of polysaccharides extracted from Lentinus edodes mycelia and their protective effects on INS-1 cells / Atividade antioxidante de polissacarídeos extraídos de Lentinus edodes micélio e seus efeitos protectores em células INS-1

The disruption of the delicate balance between the generation of reactive oxygen species (ROS) and antioxidant scavenging systems can lead to many health problems. Polysaccharides extracted from fungi fruit body and myceliums have several potential health benefits. In this study, the antioxidant capacity and cell protective effects of a polysaccharide isolated from Lentinus edodes mycelia (LMP) were investigated. The antioxidant properties of LMP were screened using radical scavenging (1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion), reducing power and inhibition of lipid peroxidation assays. Intracellular reactive oxygen species (ROS), in response to H2O2, was determined by the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay. Rat insulinoma (INS-1) cells apoptosis was detected using flow cytometry analysis. The results showed that LMP exhibited significantly strong radical scavenging activity. At the concentration of 2.0 mg mL-1, DPPH radical, hydroxyl radical, and superoxide anion radical scavenging rates are 80.32 ± 2.58%, 92.56 ± 3.11% and 93.73 ± 2.82%, respectively. The reducing power and inhibition of lipid peroxidation activity of LMP were as strong as the positive control. LMP alleviated the intracellular reactive oxygen species level and inhibited islet cell apoptosis significantly. The results open perspectives for studies of LMP in the treatment of diabetes.

O estresse oxidativo tem uma função biológica importante em muitos organismos. A ruptura do equilíbrio delicado entre a geração de espécies reativas de oxigênio (ROS) e sistemas de eliminação de antioxidantes pode levar a muitos problemas de saúde. Polissacarídeos extraídos do corpo fungos frutas e micélio têm vários benefícios potenciais para a saúde. Neste estudo, foram investigados os efeitos celulares e capacidade de proteção antioxidante de um polissacarídeo isolado a partir de Lentinus edodes micélios (LMP). As propriedades antioxidantes da LMP foram rastreadas utilizando eliminação de radicais (1, 1-difenil-2-picrilhidrazil (DPPH), hidroxilo e anião superóxido), e a redução da potência de inibição de ensaios de peroxidação lipídica. Espécies reativas de oxigénio intracelulares (ROS), em resposta ao H2O2, foi determinada pelo ensaio de diacetato de dicloro-di-hidro-f luoresceina (DCFH-DA). INS-1 células a apoptose foi detectada utilizando análise de citometria de fluxo. Os resultados mostraram que LMP exibiu significativamente forte atividade de eliminação de radicais. Na concentração de 2 mg mL-1, DPPH radical, radical hidroxilo e anião superóxido velocidades de eliminação de radicais livres são 80,32 ± 2,58%, 92,56 ± 3,11% 93,73 ± 2,82%, respectivamente. O poder redutor e inibição da peroxidação lipídica de LMP foram tão forte quanto o controlo positivo. LMP aliviou a nível de espécies reativas de oxigênio intracelular e apoptose de células da ilhota inibiu significativamente. Sugere-se que LMP pode ser utilizado como um reagente complementar para a diabetes a prevenção e terapia.

Isolation and characterization of an antifungal peptide from fruiting bodies of edible mushroom Lentinus squarrosulus Mont

Aims: To isolate and characterize an antimicrobial peptide from fruiting bodies of Lentinus squarrosulus Mont., the Thai common edible mushroom. Methodology and results: Solid ammonium sulfate at 40-80% (w/v) final concentration was utilized to precipitate the proteins and further purified by ion exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephadex G-25. The peptide was adsorbed on DEAE-cellulose and Sephadex G-25. It appeared as a single band with a molecular mass of about 17 kDa (kilodalton) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Further investigation of antimicrobial properties of purified peptide revealed that it has no activity against both Gram positive and Gram negative bacteria. However, it exhibited strong antifungal activity against various species of fungal pathogen of human. Among the high sensitive strains, Trichophyton mentagrophytes, T. rubrum, and Candida tropicalis are clinical isolates. Moreover, the potency was found to be concentration dependent and comparable with Ketoconazole, the commercial antifungal drug. Conclusion, significance and impact study: In this work, the novel bioactive peptide from fruiting bodies of L. squarrosulus Mont. has been isolated. It shows potent activity against various clinical isolates of fungal pathogen of human. It may have potential for pharmaceutical application.

In vitro anti-HIV-1 activity of the enzymatic extract enriched with laccase produced by the fungi Ganoderma sp. and Lentinus sp / Actividad anti-VIH-1 in vitro del extracto enzimático enriquecido con lacasa producido por los hongos Ganoderma sp. y Lentinus sp

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.

Assessment of Palm Press Fibre and Sawdust-Based Substrate Formulas for Efficient Carpophore Production of Lentinus squarrosulus (Mont.) Singer

Development of efficient substrate formulas to improve yield and shorten production time is one of the prerequisites for commercial cultivation of edible mushrooms. In this study, fifteen substrate formulas consisting of varying ratios of palm press fibre (PPF), mahogany sawdust (MS), Gmelina sawdust, wheat bran (WB), and fixed proportions of 1% calcium carbonate (CaCO3) and 1% sucrose were assessed for efficient Lentinus squarrosulus production. Proximate compositions of mushrooms produced on the different substrate formulas were also analysed and compared. Substrate formulations containing 85% PPF, 13% WB, 1% CaCO3, and 1% sucrose were found to produce the highest carpophore yield, biological efficiency and size (206.5 g/kg, 61.96%, and 7.26 g, respectively). Days to production (first harvest) tended to increase with an increase in the amount of WB in the substrate formulas, except for PPF based formulas. The addition of WB in amounts equivalent to 8~18% in substrate formulas containing 80~90% PPF resulted in a decrease in the time to first harvest by an average of 17.7 days compared to 80~90% MS with similar treatment. Nutritional content of mushrooms was affected by the different substrate formulas. Protein content was high for mushrooms produced on formulas containing PPF as the basal substrate. Thus, formulas comprising PPF, WB, CaCO3, and sucrose at 85% : 13% : 1% : 1%) respectively could be explored as starter basal ingredients for efficient large scale production of L. squarrosulus.

Preparation of polysaccharide complex and its indirect antitumor activity in vitro / 吉林大学学报(医学版)

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.

Lentinus edodes Suppresses Allergen-Induced Airway Hyperresponsiveness and Inflammation by Downregulating Nuclear Factor-kappa B Activity in a Murine Model of Allergic Asthma / 체질인류학회지

Medicinal mushrooms have been shown to have profound health promoting benefits. Among them, Lentinus edodes is well-known to have anti-tumor, anti-inflammatory, and immunomodulatory effects. The aim of the present study is to evaluate whether Lentinus edodes ethanol extract (LE) inhibit airway inflammatory response in a murine asthma model induced by exposure to ovalbumin (OVA). The pretreatment of LE substantially attenuated airway hyperresponsiveness and eosinophilic inflammation in OVA-challenged mice. In addition, the increased levels of Th2 cytokines (IL-4, IL-5, and IL-13), eotaxin, and adhesion molecules in bronchoalveolar lavage fluids at 48 h after OVA inhalation was significantly reduced by the administration of LE. Furthermore, LE suppressed OVA-induced activation of nuclear factor-kappa B and p38 mitogen-activated protein kinase in lung tissues. Taken together, it is proposed that LE may serve as an effective therapeutic agent for allergic airway disease.

Review on chemistry and anti-tumor effect of lentinan / 中国药学杂志

OBJECTIVE: To introduce the progress on the study of lentinan chemistry and anti-tumor effect and provide references for research and clinical applications. METHODS: Reviewed and analyzed many literatures of lentinan, summarized them compres-sively and introduced the progress on the extraction and purification of lentinan, structural analysis and anti-tumor effect in recent years. RESULTS: As the extraction and purification methods for lentinan were different, the polysaccharide compositions and structures were different, and the main forms were homopolysaccharide(glucan) and heteropolysaccharide. And its anti-tumor effects were related to immune enhancement, inhibition of tumor growth effect, synergistic anti-tumor chemotherapy and so on. CONCLUSION: Chemical properties of lentinan is closely related with the anti-tumor effect, and further in-depth research on the structure-activity relationships of lentinan is needed to provide better scientific basis for the comprehensive development and application of lentinan.

Crescimento micelial de Lentinus sajor caju (Fr.) Fr. E Pleurotus spp. em diferentes resíduos agrícolas / Mycelial growth of Lentinus sajor caju (Fr.) Fr. and Pleurotus spp. in different agricutural wastes

O objetivo deste trabalho foi avaliar o desenvolvimento in vitro de três linhagens de cogumelos [Pleurotus ostreatoroseus Singer, Pleurotus pulmonarius (Fr.) Quél. e Lentinus sajor caju (Fr.) Fr.], reativados em meio de cultura BDA e cultivadas em meios de cultura formulados à base de resíduos agrícolas, como a palha de arroz, casca de mamona, casca de amendoim. No meio sólido formulado foi adicionado um disco de cultura no centro da placa, estas foram incubadas a 25oC até a colonização do meio. Avaliou-se, diariamente, o diâmetro da colônia e obteve-se, aos cinco dias de cultivo, a massa miceliana, sendo os resultados obtidos submetidos a análise de variância e teste de Tukey. O meio contendo extrato da casca de amendoim foi o mais adequado para o crescimento da linhagem utilizada de Lentinus sajorcaju, que colonizou 68,30% da placa de Petri. O meio contendo extrato de casca de mamona foi mais favorável ao crescimento de Pleurotus ostreatoreoseus que colonizou 40,01% da placa. A linhagem nativa de Pleurotus pulmonarius cresceu indiferentemente nos meios testados e teve maior crescimento comparada a linhagem de Pleurotus ostreatoroseus. A linhagem de Lentinus sajor-caju no meio contendo extrato da casca de amendoim, apresentou massa micelial seca significativamente maior que todas as demais. O crescimento micelial das linhagens de Lentinus sajor-caju e Pleurotus ostreatoroseus é influenciado pelo meio de cultivo, enquanto o da linhagem nativa utilizada não é influenciado pelos meios testados.

The aim of this study was evaluate the in vitro development of three mushrooms strains [Pleurotus ostreatoroseus Singer, Pleurotus pulmonarius (Fr.) Quél. e Lentinus sajor-caju (Fr.) Fr.], growing media based in agricultural wastes, such rice straw, castor bean seed husks and peanut shells. In the solid medium was added a culture disc in the center of a petri dish and than was incubated at 25oC even the completly colonization of the medium, the data obtained were submitted to variance analysis and Tukey test. Was evaluated, daily, the colony diameter and the mycelial mass obtained after five days of culture. The medium containing peanut husks extract provides the optimal development for the Lentinus sajor-caju strain, when an average of 68.3% of each petri dishes was colonized by mycelia. The medium containig castor bean seed shells were most favorable for Pleurotus ostreatoreoseus development, such colonizing 40,01% of each petri dishes. The wild strain of Pleurotus pulmonarius grew indifferently in the tested media and reaches the higher development when compared with Pleurotus ostreatoroseus strain. Lentinus sajor-caju demonstrate significantly more mycelial dry mass in the peanut shells medium when compared with the others strains. This result suggests that the mycelial growing of Lentinus sajor-caju and Pleurotus ostreatoroseus are influencied by the media. For the wild strain the mycelial growing its not significantly influencied by the tested media.

Shiitake Mushroom: A Tool of Medicine.

Medicinal mushrooms have an established history of use in traditional oriental therapies. Contemporary research has validated and documented much of the ancient knowledge. Over the last three decades, the interdisciplinary fields of science that study medicinal mushrooms has sprung up and has increasingly demonstrated the potent and unique properties of compounds extracted from a range of species. Currently, the field is being developed into a very fruitful area. Modern clinical practice in Japan, China, Korea and other Asian countries rely on mushroom-derived preparations. Mushrooms have been studied for nutritional and medical purposes for its various potential anti-tumoral and immunomodulatory componests like polysaccharides that have been identified. For medical purposes, mushrooms have been consumed to prevent cancer and cardiac diseases, to improve blood circulation and to reduce blood cholesterol level. Some of these mushrooms have also been used for the treatment of physical and emotional stress, osteoporosis, gastric ulcers and chronic hepatitis, for the improvement of the quality of life of patients with diabetes and especially for the stimulation of immunity. Shiitake has a history of medicinal uses. The mushroom is used as anticarcinogenic, antiinflammatory, antioxidant, antifungal, antibacterial, antiviral as well as antithrombotic in cardiovascular disorders. This article has been written to throw some light on Shiitake mushroom which has many nutritional values. Many Shiitake preparations came in market containing the active ingredients which can replace many other marketed synthetic medicines and may prove to have promising results with fewer side effects.

Isolation of a laccase with HIV-1 reverse transcriptase inhibitory activity from fresh fruiting bodies of the Lentinus edodes (Shiitake mushroom).

A laccase with a molecular mass of 67 kDa and inhibitory activity toward HIV-1 reverse transcriptase (IC50 = 7.5 M) was isolated from fresh fruiting bodies of the Lentinus edodes (Shiitake mushroom). Its characteristics were compared with those of laccases from cultured mushroom mycelia reported earlier. The laccase was unadsorbed on DEAE-cellulose, Affi-gel blue gel and CM-cellulose, but was adsorbed on Con A-Sepharose. About 50-fold purification was achieved with a 19.2% yield of the enzyme. The activity of the enzyme increased steadily from 20°C to 70°C. The activity disappeared after exposure to the boiling temperature for 10 min. Its optimal pH was 4 and very little enzyme activity remained at and above pH 10. The laccase inhibited HIV-1 reverse transcriptase with an IC50 of 7.5 M, but did not demonstrate any antifungal or anti-proliferative activity.

Protective Effects of a Water-Soluble Extract from Culture Medium of Lentinus Edodes Mycelia against Neuronal Damage after Hypoxia-Ischemia in Mice / 日本補完代替医療学会誌

<b>Objective</b>: <i>Lentinus edodes</i> (Shiitake) is a very popular mushroom in Asian cuisine. The water-soluble extract from culture medium of <i>Lentinus edodes</i> mycelia (LEM), which is commercially available as a nutritional supplement, is prepared by hot-water treatment from a solid medium composed of bagasse and defatted-rice bran overgrown for about 4 months with its mycelia. LEM was previously reported to have antioxidant activity and to suppress various oxidative damages. In this study, the neuroprotective effects of 2-week intake of LEM on cerebral ischemic damage induced by hypoxia/ischemia (H/I) followed by reoxygenation in mice were examined.<br> <b>Method</b>: Male C57BL/6J mice were divided into three groups, fed for two weeks with the control laboratory powder chow, 0.5% LEM-contained chow, or 1% LEM-contained chow, respectively. Cerebral ischemic damage was induced in the mice by H/I (i.e., unilateral ligation of the carotid artery and exposure of 8%O₂ for 30 min). Twenty-four hours after H/I, total plasma oxidative stress, neurological deficits, cerebral infarction volume were evaluated in each group. Furthermore, the number of apoptotic cells in ischemic penumbra, the hippocampal CA1 and CA2, and the somatosensory area of the cortex, were analyzed by TUNEL staining and cleaved caspase-3 immunostaining.<br> <b>Results</b>: The infarct area assessed 24-h after H/I was extended to the corpus striatum and cortex in the control mice. Treatment of LEM dose dependently improved plasma oxidative stress, neurological deficits, and cerebral infarction volume. Moreover, LEM decreased the levels of dihydroethidium activity as an index of super oxide production and the number of apoptotic cells in ischemic penumbra.<br> <b>Conclusion</b>: These results show that chronic intake of LEM relieves the hypoxia-induced cerebral ischemic injury, which may be attributed to the antioxidant effects of LEM.<br>

Appraisal of Antihyperlipidemic Activities of Lentinus lepideus in Hypercholesterolemic Rats

The wild edible mushroom, Lentinus lepideus has recently been cultivated for commercial use in Korea. While the mushroom has been widely used for nutritional and medicinal purposes, the possible anti-hyperlipidemic action is unclear. The effects of dietary L. lepideus on plasma and feces biochemical and on the liver histological status were investigated in hypercholesterolemic rats. Six-wk-old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Biochemical and histological examinations were performed. A diet containing 5% L. lepideus fruiting bodies reduced plasma total cholesterol, triglyceride, low-density lipoprotein, total lipid, phospholipids, and the ratio of low-density to high-density lipoprotein. Body weight was reduced. The diet did not adversely affect plasma biochemical and enzyme profiles. L. lepideus reduced significantly plasma beta- and pre-beta-lipoprotein, while alpha-lipoprotein content was increased. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. The present study suggests that a diet supplemented with L. lepideus can provide health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.

Antihyperlipidemic Effect of Dietary Lentinus edodes on Plasma, Feces and Hepatic Tissues in Hypercholesterolemic Rats

We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.

In vitro activity of hypnophilin from Lentinus strigosus: a potential prototype for Chagas disease and leishmaniasis chemotherapy

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.

Lentinula edodes (Shiitake) mushroom extract protects against hydrogen peroxide induced cytotoxicty in peripheral blood mononuclear cells.

Lentinula edodes (Berk) Pegler, commonly known as Shiitake mushroom has been used as medicinal food in Asian countries, especially in China and Japan and is believed to possess strong immunomodulatory property. In the present study, the methanolic extract of the fruit bodies of L. edodes was investigated for cytoprotective effect against H2O2-induced cytotoxicity in human peripheral blood mononuclear cells (PBMCs) by measuring the activities of xanthine oxidase (XO) and glutathione peroxidase (GPx) . H2O2 at a concentration of 5 μM caused 50% inhibition of PBMCs viability. The extract improved the PBMC viability and exerted a dose-dependent protection against H2O2-induced cytotoxicity. At 100 μg/ml of extract concentration, the cell viability increased by 60% compared with the PBMCs incubated with H2O2 alone. The extract also inhibited XO activity in PBMC, while showing moderate stimulatory effect on GPx. However, in the presence of H2O2 alone, both the enzyme activities were increased significantly. The GPx activity increased, possibly in response to the increased availability of H2O2 in the cell. When the cells were pretreated with the extract and washed (to remove the extract) prior to the addition of H2O2, the GPx and XO activities as well as the cell viability were comparable to those when incubated with the extract alone. Thus, it is suggested that one of the possible mechanisms via which L. edodes methanolic extract confers protection against H2O2-induced oxidative stress in PBMC is by inhibiting the superoxide-producing XO and increasing GPx activity which could rapidly inactivate H2O2.

The Suppressive Effect of Lentinus Edodes on Renal Scarring Secondary to Pyelonephritis / 대한비뇨기과학회지

PURPOSE: Lentinus edodes is used in both foods and drugs, and is also known to be an antioxidant agent. Some investigators have noted that renal scarring following pyelonephritis is closely related to inflammation or free oxygen radicals, rather than to direct injury due to bacterial infection. The efficiency of the antioxidant agent, lentinus edodes, in preventing the renal scarring caused by experimental pyelonephritis was evaluated in rats. MATERIALS AND METHODS: Twenty eight male Sprague Dawley rats (n=28) were injected into both renal parenchyma with 0.1ml of a solution containing Escherichia coli (10(8)/ml). The control group, consisting of 4 rats, received an injection of isotonic saline instead of the bacterial solution. Three days after surgery, the animals were given the following treatments: in the pyelonephritis group, 7 rats received isotonic saline for 5 days; in the antibiotics treatment group, 6 rats were treated with an antibiotic only, ciprofloxacin, for 5 days. In the lentinus edodes treatment group, 5 rats were treated with lentinus edodes only. In the combined group, 6 rats received both lentinus edodes and ciprofloxacin. The kidneys were harvested 6 weeks after infection, and histopathologically examined for renal scarring. RESULTS: Delayed treatment with antibiotics-only or lentinus edodes-only had no effect on the scarring compared with the untreated controls. However, the addition of lentinus edodes to the delayed antibiotic therapy significantly inhibited the renal scarring compared with the pyelonephritis, antibiotic-only treated and the lentinus edodes-only treated groups (p<0.05). CONCLUSIONS: The administration of lentinus edodes in combination with antibiotics significantly reduced the renal scar formation in pyelonephritis.

Supplementary Effect of Lentinus Edodes on Serum and Hepatic Lipid Levels in Spontaneously Hypertensive Rat

This study was conducted to investigate the supplementary effects of Lentinus edodes on blood pressure and blood lipid levels in the Hypertensive Rat. We supplied 3 kinds of experimental diets (Control; CO, cap of Lentinus edodes; LC, and stipe of Lentinus edodes; LS) to spontaneously hypertensive rats (SHR) for 8 weeks. Diet intake, body weight, organ weights, and serum lipid levels were measured. There was no significant difference in diet intake, and body, liver and epididymal fat pad weights among experimental groups. The levels of systolic and diastolic blood pressure were significantly lower in LC and LS groups than CO group. The concentrations of serum total cholesterol were lower in LC and LS groups than CO group. These results suggested that Lentinus edodes decrease blood pressure levels and serum total cholesterol concentration in the spontaneously hypertensive rats. But these effects of Lentinus edodes didn't show any significant difference between animals fed cap and stipe of this mushroom.