An insight into antidiabetic properties of six medicinal and edible mushrooms: Inhibition of α-amylase and α-glucosidase linked to type-2 diabetes

As a continuation of our search for biologically active mushroom species the present study investigates in vitro antidiabetic properties of six edible and medicinal mushroom species: Agaricus blazei Murrill, Coprinus comatus (O.F.Müll.) Pers., Cordyceps militaris (L.) Fr., Inonotus obliquus (Ach. ex Pers.) Pilát, Morchella conica Pers. and Phellinus linteus Berk. & M.A. Curtis. In vitro assays on α-amylase and α-glucosidase enzyme inhibition were performed with methanolic extracts of the selected mushrooms. Furthermore, we calculated the necessary daily intake of mushroom extracts and dry mushroom powders based on the equivalent doses of therapeutic drug acarbose given to diabetic patients per day. Our comparative study on enzyme inhibition showed that the most promising potential is ascribed to I. obliquus extract, while no inhibition of α-amylase was recorded with M. conica and C. militaris methanolic extract at the tested concentration. This comparative study is the first highlighting in vitro antidiabetic potential by inhibition of α-amylase and α-glucosidase with methanolic extracts; which makes the investigated speciesmore promising for the diabetes type-2 treatment by an additional and different mechanism of action.The authors thank Serbian Ministry of Education, Science and Technological Development for the financial support (grant number 173032).info:eu-repo/semantics/publishedVersio

A study on the medicinal mushroom Cordyceps militaris (L.) link: chemical characterization, antioxidant, antimicrobial and antiproliferative properties

The Cordyceps genus is well known for its medicinal properties. A number of bioactive constituents from Cordyceps species have been reported such as antibacterial, antifungal, immunopotentiating or antitumor agents [1]. The anti-inflammatory and anti-angiogenic properties of Cordyceps militaris (L.) Link have been reported as also the antioxidant activity of its cultured mycelium [2]. In this work, the chemical characterization of C. militaris was performed, includingbioactive compounds (free sugars, unsaturated fatty acids, tocopherols, organic acidsand phenolic compounds). The antioxidant potential of its methanolic extract was evaluated (reducing power, scavenging activity and lipid peroxidationinhibition) as also the antimicrobial activity (tested towards Gram positive and Gram negative bacteria and eight microfungi) and antiproliferative properties (tested in different human tumor cell lines). Mannitol and trehalose were the main free sugars found in this species (2.01 and 24.71 g/100 g dw, respectively). The fatty acid quantified in higher amounts was linoleic acid (68.00% of total fatty acids). a-Tocopherol was the only isoform of vitam in E detected (55.86 j.lg/1 00 g dw). The organic acids found in this mushroom were oxalic (0.33 g/1 00 g dw), citric (7.97 g/1 00 g dw) and fumaric (0 .1 3 g/100 g dw) acids. p-Hydroxybenzoic acid was the only phenolic acid quantified in C. militaris (0.02 mg/1 00 g dw), but cinnamic acid was also found (0. 11 mg/1 00 g dw). The lowest EC50 values for the antioxidant potential were presented in lipid peroxidation inhibition assays, namely in 13-carotene/l inoleate and TBARS assays (1.05 mg/ml and 0. 77 mg/ml, respectively). The methanolic extract also revealed strong antibacterial and antifungal activities, and it was able to inhibit the proliferation of MCF-7 (breast}, NCI-H460 (non-smalllung),HCT-15 (colon) and Hela (cervical) human carcinoma cell lines.FCT (Portugal) and COMPETE/QREN/EU: projects PTOC/AGR-ALI/110062/2009 and PEstOE/AGR/UI0690/2011, and to Serbian Ministry of Education and Science for financial support (grant number 173032)

The methanolic extract of Cordyceps militaris (L.) Link fruiting body shows antioxidant, antibacterial, antifungal and anti human tumor cell lines properties

Being Cordyceps militaris (L.) Link recognized as a medicinal and edible mushroom, this work intends to reveal new interesting bioactive molecules that could be isolated from this species. Hydrophilic and lipophilic compounds were analysed by chromatographic techniques coupled to different detectors. The methanolic extract of C. militaris was tested for its antioxidant, antibacterial, antifungal and anti-proliferative properties in different human tumor cell lines. Mannitol (2.01 g/100 g dw) and trehalose (24.71 g/100 g) were the free sugars found in C. militaris. Polyunsaturated fatty acids (68.87%) predominated over saturated fatty acids (23.40%) and δ-tocopherol was the only isoform of vitamin E detected (55.86 μg/100 g). The organic acids found in this mushroom were oxalic, citric and fumaric acids (0.33, 7.97 and 0.13 g/100g, respectively). p-Hydroxybenzoic acid was the only phenolic acid quantified in this species (0.02 mg/100 g); although cinnamic acid was also found (0.11 mg/100 g). The methanolic extract of C. militaris proved to inhibit lipid peroxidation, have reducing power and scavenge free radicals. This extract also revealed strong antibacterial and antifungal properties. Finally, the C. militaris extract was able to inhibit the proliferation of MCF-7 (breast), NCI-H460 (non-small lung), HCT-15 (colon) and HeLa (cervical) human carcinoma cell lines.Foundation for Science and Technology (FCT, Portugal) and COMPETE/QREN/EU for financial support to this work (research project PTDC/AGRALI/ 110062/2009 and SFRH/BPD/BPD/68344/2010 to R. Calhelha), to CIMO (strategic project PEst-OE/AGR/UI0690/2011) and to L. Barros researcher contract under “Programa Compromisso com Ciência-2008”). The authors also thank to Serbian Ministry of Education and Science for financial support (grant number 173032)

Nutrients and non-nutrients composition and bioactivity of wild and cultivated Coprinus comatus (O.F.Müll.) Pers.

Mushrooms have been reported as sources of biomolecules with various potential. Coprinus comatus was studied to obtain information about this species, comparing cultivated and wild samples. Free sugars, fatty acids, tocopherols, organic acids and phenolic acids were analysed by chromatographic techniques coupled to different detectors. C. comatus methanolic extractwas tested for its antioxidant potential (reducing power, radical scavenging activity and lipid peroxidation inhibition) and antimicrobial properties (tested towards Gram positive and negative bacteria, and microfungi). The toxicity for liver cells was tested in porcine liver primary cells. Both studied samples revealed similar nutritional value and energy contribution. The cultivated sample revealed the highest content in free sugars, monounsaturated fatty acids and tocopherols, while the wild mushroom was richer in saturated and polyunsaturated fatty acids, organic acids and phenolic compounds. The cultivated species also revealed the highest antioxidant potential and antimicrobial activity (with exception towards Gram negative bacteria and Aspergillus ochraceus). Both species revealed no toxicity towards porcine liver cells. The present study proved that cultivated and wild mushrooms from the same species could be excellent options as food and as sources of nutritional and bioactive compounds. Furthermore, differences in wild and cultivated samples were comparatively investigated for the first time.Fundação para a Ciência e Tecnologia (FCT, Portugal) and COMPETE/QREN/EU for financial support to this work (research project PTDC/AGR-ALI/110062/2009 and BPD/4609/2008 to L. Barros) and to CIMO (strategic project PEst-OE/AGR/UI0690/2011). The authors also thank Serbian Ministry of Education and Science for financial support (grant number 173032)

Cultivated strains of Agaricus bisporus and A. brasiliensis: chemical characterization and evaluation of antioxidant and antimicrobial properties for final healthy product - natural preservatives in yoghurt

Agaricus bisporus (J. E. Lange) Emil J. Imbach and Agaricus brasiliensis Wasser, M. Didukh, Amazonas & Stamets are edible mushrooms. We chemically characterized these mushrooms for nutritional value, hydrophilic and lipophilic compounds. The antioxidant and antimicrobial activities of methanolic and ethanolic extracts were assessed. Hepatotoxicity was also evaluated. The ethanolic extract of both species was tested for inhibition of Listeria monocytogenes growth in yoghurt. Both species proved to be a good source of bioactive compounds. A. brasiliensis was richer in polyunsaturated fatty acids and revealed the highest concentration of phenolic acids, and tocopherols. A. bisporus showed the highest monounsaturated fatty acids and ergosterol contents. A. brasiliensis revealed the highest antioxidant potential, and its ethanolic extract displayed the highest antibacterial potential; the methanolic extract of A. bisporus revealed the highest antifungal activity. A. brasiliensis possessed better preserving properties in yoghurt

A detailed comparative study between chemical and bioactive properties of Ganoderma lucidum from different origins

A detailed comparative study on chemical and bioactive properties of wild and cultivated Ganoderma lucidum from Serbia (GS) and China (GCN) was performed. This species was chosen because of their worldwide use as medicinal mushroom. Higher amounts of sugars were found in GS, while higher amounts of organic acids were recorded in GCN. Unsaturated fatty acids predominated over saturated fatty acids. GCN revealed higher antioxidant activity, while GS exhibited inhibitory potential against human breast and cervical carcinoma cell lines. No cytotoxicity in non-tumour liver primary cell culture was observed for the different samples. Both samples possessed antibacterial and antifungal activities, in some cases even better than the standard antimicrobial drugs. This is the first study reporting a comparative of chemical compounds and bioactivity of G. lucidum samples from different origins

Chemical characterization of the medicinal mushroom Phellinus linteus (Berkeley & Curtis) Teng and contribution of different fractions to its bioactivity

Mushrooms are widely appreciated for their organoleptic qualities, being also recognized as good sources of bioactive compounds that provide antioxidant, antimicrobial and cytotoxic activities. Polysaccharides (including glucans) are often pointed out as the most bioactive compounds isolated from mushrooms, but other molecules such as triterpenoids, might also be highlighted for their bioactivity. In scientific research, when isolated compounds are used, potential synergistic effects might be lost. Accordingly, the bioactivity of Phellinus linteus was evaluated in selected fractions (polysaccharides, glucans and triterpenoids), as well as in the methanolic and ethanolic extracts. The best antioxidant and antibacterial activities were obtained with methanolic extract, while glucan and triterpenoid fractions gave the strongest antifungal activity. In contrast, ethanolic extract gave the best results in cytotoxic activity, indicating that the bioactive compounds present might act synergistically. The differentiated activity of P. linteus fractions and extracts could be useful to find antimicrobial, antioxidant and cytotoxic agents as alternatives to synthetic chemicals with application in agriculture, food industry or pharmacy.Foundation for Science and Technology (FCT, Portugal) and COMPETE/QREN/EU for financial support to this work (research project PTDC/AGRALI/110062/2009, BPD/72802/2010 to J.C.M. Barreira and SFRH/BPD/68344/2010 to T.C. Calhelha) and to CIMO (strategic project PEst-OE/AGR/UI0690/2011). The authors also thank to Serbian Ministry of Education and Science for financial support (grant number 173032). Phellinus linteus fruiting body powder, crude polysaccharides and triterpenoids were kind gifts of Amazing Grace Health Industries, Bangkok, Thailand

Chemical characterization and biological activity of Chaga (Inonotus obliquus), a medicinal “mushroom”

Chemical composition and biological properties of aqueous and ethanolic extracts of Inonotus obliquus (Pers.:Fr.) Pilat from different origins, i.e. of Finland, Russia, and Thailand, were studied. Concerning biological activity, antimicrobial, antiqourum, antioxidative, and antitumor and cytotoxic effects were tested. Oxalic acid was found as the main organic acid, with the highest amount in Russian aqueous extract. Gallic, protocatechuic and p-hydroxybenzoic acids were detected in all samples. I. obliquus extracts showed high antioxidant and antimicrobial activity. Extracts were tested at subMIC for anti-quorum sensing (AQS) activity in Pseudomonas aeruginosa and all of them showed definite AQS activity. The assays were done using twitching and swarming of bacterial cultures, as well as the amount of produced pyocyanin as QS parameters. All the extracts possessed good antitumor effect on four cells lines and did not show hepatotoxicity on porcine liver primary cells designed as PLP2. As the I. obliquus presence in Chaga conks is limited, further purification is necessary to draw quantitative conclusions. The presence of AQS activity in medicinal mushrooms suggests a broader anti-infectious disease protection than only immunomodulatory effects.The authors are grateful to Serbian Ministry of Education, Science and Technological Development for financial support (grant number 173032), to Foundation for Science and Technology (FCT, Portugal) and COMPETE/QREN/EU for financial support to CIMO strategic project PEst-OE/AGR/UI0690/2011, L. Barros contract, A. Fernandes (SFRH/BD/76019/2011) and R.C. Calhelha (SFRH/BPD/68344/2010) grants

Real-time monitoring of IL-6 and IL-10 reporter expression for anti-inflammation activity in live RAW 264.7 cells

In previous study, we suggested that the interleukin (IL)-6 and IL-10 could serve as a good biomarker for anti-inflammation that related to chronic inflammatory disease. Recently, we are finding new anti-inflammation compounds from natural products by screening of IL-6 and IL-10 levels. Although, we could measure IL-6 and IL-10 levels by several methods. However, all methods could not measure continuous kinetic of IL-6 and IL-10 levels. Most methods have multiple steps and take a long time. Therefore, there is no a suitable method for screening. To this end, we established IL-6 and IL-10 promoter assay which can monitor with reference gene as Glyceraldehyde 3-phosphate dehydrogenase (gapdh) promoter in living single cell. It could determine IL-6 and IL-10 levels continuously in real-time within two steps. We evaluated IL-6 and IL-10 reporter expression in LPS-induced RAW 264.7 cells with well-known anti-inflammatory compounds such as quercetin, xanthones, β-D-glucan and dexamethasone. As the results, the expression of IL-6 and IL-10 reporters were strongly induced by LPS. The expression of IL-6 reporter was inhibited by all anti-inflammation compounds in LPS-induced RAW 264.7 cells. The expression of IL-10 reporter was inhibited by quercetin, xanthones and dexamethasone in LPS-induced RAW 264.7 cells. While, expression of IL-10 reporter was induced by β-D-glucan. These results indicated that this assay could use for determination of IL-6 and IL-10 reporter expression in LPS-induced RAW 264.7 cells for anti-inflammation activity. Moreover, the results showed that natural compounds have an effect on the time course of IL-6 and IL-10 expressions. Therefore, real-time monitoring has a merit for natural compounds screening. We suggested that this assay could serve as a compound screening assay for anti-inflammation activity