Portuguese Wild Mushrooms: Chemical characterization and functional study of antiproliferative and proapoptotic properties in cancer cell lines

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Antioxidants in Pinus Pinaster roots and mycorrhizal fungi during the early steps of symbiosis

Publication . Carocho, Márcio; Barros, Lillian; Ferreira, Isabel C.F.R.; Barreira, João C.M.; Martins, Anabela

Ectomycorrhizal symbiosis between fungi mycelia and the roots of some plants could have important effects in the levels of antioxidants of both partners. In the present work, the effects of co-culture period in the antioxidant properties and antioxidants (phenolics, tocopherols and sugars) production during early steps of in vitro mycorrhization (Pinus pinaster-Pisolithus arhizus and Pinus pinaster-Paxillus involutus) were evaluated. The studied parameters were determined in each culture element (root, mycelium and medium) in order to understand the response of each partner to the symbiotic association. P. arhizus proved to be more compatible with P. pinaster than P. involutus, since the antioxidant activity in the latter species increased from 48 to 72 h, while P. arhizus antioxidant properties decreased at the same period (indicative of less oxidative stress). Despite P. involutus proved to be less suitable to be included in forestation programs using mycorrhization processes, it revealed a higher potential for bioactive compounds production in the early steps of symbiosis. A maximal value of phenolics content was obtained after the first 6 h.

2012Journal article

Open access

Antioxidant properties of two Lactarius wild species from the Northeast of Portugal

Publication . Vieira, Vanessa; Reis, Filipa S.; Martins, Anabela; Ferreira, Isabel C.F.R.

Wild mushrooms are widely appreciated not only for their organoleptic and nutritional properties, but also for their medicinal potential due to the accumulation of bioactive compounds. They have been described as a source of many antioxidant compounds such as phenolic compounds, tocopherols, ascorbic acid and carotenoids (1]. In the present work, the antioxidant properties of two wild Lactarius species from the Northeast of Portugal (Lactarius citroleus and Lactarius plumbeus) were evaluated for the first time, allowing the comparison between these two species. The antioxidant properties were determined through five different assays. The reducing power of the samples was assessed through the Folin-Ciocalteu (or total phenolics) and the Ferricyanide/Prussian blue assays. To evaluate the radical scavenging activity, the DPPH-2,2- diphenyl-1-picrylhydrazylassay was applied. Lipid peroxidation inhibition was measured through two different assays: β-carotene/linoleate (or β-carotene bleaching inhibition) and TBARSthiobarbituric- acid-reactive-species assays. Although L. plumbeus has shown the highest content in total phenolics (22.02 mg of gallic acid equivalents/g extract), L. citroleus presented the highest reducing power determined by the Ferricyanide/Prussian blue assay (EC50=1 .53 mg/ml ). L. plumbeus revealed the highest radical scavenging activity (EC50=4.18 mg/ml) as also the highest lipid peroxidation inhibition measured by β-carotene/linoleate assay (EC50=4.92 mg/ml). Nevertheless the lowest EC50 value for TSARS assay was revealed by L. citroleus (0.57 mg/ml). This study contributes for the inventorying of the wild mushroom species from the Northeast of Portugal, providing more information about the antioxidant potential of these natural matrices.

2013Conference object

Open access

An overview of the nutrients and non-nutrients present in the wild mushroom species most appreciated in the Northeast of Portugal

Publication . Fernandes, Ângela; Oliveira, Beatriz; Martins, Anabela; Ferreira, Isabel C.F.R.

Our research group has an extensive work in the chemical characterization of wild mushroom species from the Northeast of Portugal. This region is one of the European regions with higher wild edible mushrooms diversity, some of them with great gastronomic relevance. In Europe, they are collected for consumption being a good source of digestible proteins, carbohydrates, fibers and vitamins. The present work reports the nutrients {fatty acids and sugars) and non-nutrients {phenolic acids) present on the wild species most appreciated in the Northeast of Portugal: Boletus edulis, Cantharellus cibarius, Lactarius deliciosus and Macrolepiota procera. Fatty acids and sugars were obtained by GC-FID and HPLC-RI, respectively [1-3]; Phenolic acids were analysed by HPLC-DAD-ESI/MS [4]. Furthermore their nutritional value was accessed through the composition in macronutrients and energetic value. Boletus edulis gave the highest energetic contribution and MUFA {mainly oleic acid) content; Cantarellus cibarius revealed the highest moisture and proteins levels; Lactarius deliciosus showed the highest ash, fat, SFA {mainly due to the specific contribution of stearic acid), sugars (total contribution of mannitol and trehalose) and phenolic acids {mainly p-hydroxybenzoic acid) contents; Macrolepiota procera gave the highest levels of total carbohydrates and PUFA. It was concluded that the most appreciated wild mushrooms have different nutrients and non-nutrients profiles and concentrations. This report contributes to the elaboration of nutritional data bases of the most consumed species, allowing comparison between them.

2012Conference object

Open access

Antimicrobial activity of phenolic compounds identified in wild mushrooms, SAR analysis and docking studies

Publication . Alves, Maria José; Ferreira, Isabel C.F.R.; Froufe, Hugo J.C.; Abreu, Rui M.V.; Martins, Anabela; Pintado, Manuela

Aim and Methods: Although the antimicrobial activity of extracts from several mushroom species have been reported, studies with the individual compounds present in that extracts are scarce. Herein, the antimicrobial activity of different phenolic compounds identified and quantified in mushroom species from all over the world was evaluated. Furthermore, a structure activity relationship (SAR) analysis and molecular docking studies were performed, in order to provide insights in the mechanism of action of potential antimicrobial drugs for resistant microorganisms. Results: 2,4-Dihydroxybenzoic and protocatechuic acids were the phenolic compounds with higher activity against the majority of Gram negative and Gram positive bacteria. Furthermore, phenolic compounds inhibited more MRSA than methicillin sensible Staphylococcus aureus. MRSA was inhibited by 2,4-dihydroxybenzoic, vanillic, syringic (MICs=0.5 mg/mL) and p-coumaric (MIC= 1 mg/mL) acids, while these compounds at the same concentrations had no inhibitory effects against methicillin sensible Staphylococcus aureus. Conclusions: The presence of carboxylic acid (COOH), two hydroxyl (OH) groups in para and ortho positions of the benzene ring, as also a methoxyl (OCH3) group in the meta position seems to be important for anti-MRSA activity. Significance and Impact of the Study: Phenolic compounds could be used as antimicrobial agents, namely against some microorganisms resistant to commercial antibiotics.

2013Journal article

Open access

A study of the antitumour potential of three Portuguese wild mushrooms

Publication . Vaz, Josiana A.; Ferreira, Diana G.; Almeida, Gabriela M.; Cardoso, Adélia; Almeida, Ana P.; Martins, Anabela; Ferreira, Isabel C.F.R.; Vasconcelos, M. Helena

Natural matrixes such as mushrooms represent a rich source of biologically active compounds with recognized potential in drug discovery and development [1,2]. Indeed, many pre-clinical studies have been conducted in human tumour cell lines and in some cases, a number of compounds extracted from mushrooms have entered clinical trials [3]. Our previous results showed that extracts from Agaricus arvensis, Suillus collinitus and Clytocibe alexandri are promising sources of low molecular weight bioactive compounds [4]. The aim of the present work was to study the antitumour potential of the extracts and isolated compounds from three Portuguese wild mushrooms by verifying their effect on various human tumour cell lines in what concerns effect on cell growth, cell cycle profile and programmed cell death. Wild mushrooms were collected from the Northeast of Portugal and classified as Agaricus arvensis, Suillus collinitus and Clitocybe alexandri. Phenolic (methanolic and ethanolic) and polissacharidic extracts were prepared. The effect of the extracts on tumour cell growth inhibition was verified with the SRB assay and the GI50 of each extract was determined for each of the cell lines studied (NCI-H460, MCF-7, AGS and HCT-15). Our preliminary results revealed that all the extracts from Clitocybe alexandri are capable of causing cell growth inhibition and provided GI50 concentrations bellow 60µg/ml in all the cell lines tested [4]. Regarding the effect of the Agaricus arvensis extracts, they all caused an inhibition of cell growth in all cell lines, particularly the methanolic extract which revealed to be a very potent inhibitor of cell growth, especially in the MCF-7 cell line. The evaluation of the effect of the Suillus collinitus extracts will be carried out as well as cell cycle and apoptosis analyses, by flow cytometry. Finally, the isolation and characterization of compounds from these extracts will also be carried out, using HPLC-DAD or HPLC-RI. The structures of the compounds will be established by NMR spectral analysis (1H, 13C, DEPT, COSY, HSQC and HMBC).

2011Conference object

Open access