Food Science and Technology 6(2): 47-55, 2018 http://www.hrpub.org DOI: 10.13189/fst.2018.060201 Beta-glucan Content, Antioxidant and Antimicrobial Activities of Some Edible Mushroom Species Özge Özcan1,*, Figen Ertan2 1Vocational College of Health Services, Kırklareli University, Kırklareli, Turkey 2Department of Biology, Edirne, Faculty of Science, Trakya University, Turkey Copyright©2018 by authors, all rights reserved. Authors agree that this article remains permanently open access under the terms of the Creative Commons Attribution License 4.0 International License Abstract In this study the β-glucan content, terpenoids, glycopeptides, saponin and ergosterol with antioxidant and antimicrobial activities of Boletus edulis, high medicinal value [3, 4]. Along with the Cantharellus cibarius, Craterellus cornucopioides, immunomodulatory and antitumour effects of the Hydnum repandum and Agaricus bisporus were studied. mushrooms, it is known that they have antioxidant, For this purpose, methanol and acetone extracts of antihypertensive, cholesterol lowering, blood glucose mushrooms were obtained. The β-glucan content of regulating, antiallergic, hypolipidemic, antiviral, mushrooms varied between 14.57 ± 0.80% - 11.26 ± antimicrobial, hepatic tissue protecting and antifibrotic 0.69%. According to the results obtained from the characteristics [5,6]. Some species of mushrooms have 2,2-diphenyl-1- picrylhydrazyl (DPPH) assay, the radical been proven to have antitumour effects [7, 8]. The scavenging activity was highest in B. edulis and A. antitumour compounds clavacin, volvotoksin, bisporus. The total phenolic content of extracts ranged flammutoksin, lentinan and porisin are isolated only from between 22.75 ± 0 .32 - 45.5 ± 0.71 µg/mg as gallic acid mushrooms [5]. equivalents. Mushroom extracts had low reducing power The most outstanding molecules among the compounds capacity in comparison to standards. When the guaiacol in the structure of mushrooms are the β-glucans. β-glucan peroxidase activity of mushrooms was examined, B. edulis is a polysaccharide in the fungal cell wall, displaying a and A. bisporus showed enzyme activities of 0.07 U/ml β(1→3) linkage and may display β(1→6) branching. It is and 0.13 U/ml, respectively. The antimicrobial activity of found in cereals, bacteria and mushrooms. β-glucan has acetone and methanol extracts of mushrooms showed effects on immune system activation as well as varied levels of inhibitory effects on the microorganisms antimicrobial, antioxidant, antiviral, antifungal, antitumour, tested. cholesterol lowering, and blood glucose regulation effects [7]. Keywords Mushroom, Antimicrobial, Antioxidant, Mushrooms synthesise secondary metabolites such as β-glucan phenolic compounds, polyketides, terpenes, and steroids. These secondary metabolites, particularly phenolic compounds found inside the mushroom, have strong antioxidant activities. Phenolic compounds function in metal chelation, free radical scavenging and lipid 1. Introduction peroxidation inhibition. Thanks to these antioxidant effects, mushrooms are effective treatments for many ailments, Mushrooms have been utilized for medicinal purposes as especially cancer and cardiac disease [9]. well as being consumed as food in many parts of the world Various antimicrobial compounds isolated from both for centuries. Mushrooms are food sources that are poor in mycelium and fruiting bodies of mushrooms have been fat while being quite rich in protein, dietary fibre, vitamins reported. These secondary metabolites function just like and minerals [1]. In the structure of the mushrooms, some antibiotics. Mushrooms need these antibiotics and vitamins minerals such as calcium, phosphorus, potassium, iron, to vegetate and reproduce. Synthetic antibiotics and copper are found apart from thiamine, riboflavin, niacin, antimicrobial medicines both threaten human health and biotin, cobalamin, ascorbic acid, vitamin D and K [2]. In cause antibiotic resistance. For this reason, new natural addition to their high nutrition content, mushrooms are rich antimicrobial substances obtained from nature are in bioactive metabolites such as lecithin, polysaccharide, necessary. In this respect, mushrooms are important phenolic and polyphenolic compounds, polyketides, organisms. Along with antibiotics, β-glucan in fungal cell 48 Beta-glucan Content, Antioxidant and Antimicrobial Activities of Some Edible Mushroom Species walls has been determined to have an antimicrobial effect method of Blois (1958) [13]. In this method, each extract [10]. was prepared at 5 different concentrations from 100-1000 The aim of this study was to determine the antimicrobial µg /ml. 1 ml from the different concentrations of these activities and β-glucan content of four different edible wild extracts was mixed with 4 ml of 0.1 mM DPPH and mushroom species that grow naturally in the Trakya vortexed. After standing for 30 minutes in the dark, the Region of Turkey and to compare them with a cultured absorbance was measured at 517 nm. Each sample was mushroom. Mushrooms were studied in the fresh form as measured in triplicate. Ethanol and methanol were used as eaten in salads, meals and canned food. negative controls and butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) as positive controls. The % DPPH radical scavenging activity was calculated 2. Materials and Methods using the formula: %DPPH radical scavenging activity 2.1. Mushrooms Control absorbance sample absorbance = x100 control absorbance The edible mushroom species were Boletus edulis (bolet − mushroom), Cantharellus cibarius (chanterelle), Craterellus cornucopioides (trumpet mushrooms) and The concentration by which a substance inhibits 50% of Hydnum repandum (anchusa mushrooms). The cultured the DPPH is expressed as IC50 [14]. The IC50 values were mushroom studied was Agaricus bisporus. Mushrooms calculated from the graph obtained from % free radical were provided and identified by Vize Mushroom Food and removal activity values against different concentrations Agricultural Products Marketing Industry and Trade studied. Limited Company and Akya Mushroom Limited Company. Mushroom samples were obtained fresh and stored at 2.4.2. Determination of total phenolic compounds -20°C until analysis. The total phenolic content soluble in methanol and acetone extracts of mushrooms was determined using Folin 2.2. Preparation of Mushroom Extract Ciocalteu Reagent (FCR) [15]. From each sample 1 mg/ml was mixed with 45 ml water and 1 ml of Folin-Ciocalteu Mushrooms stored at -20°C were allowed to stand at Reagent. After standing for 3 minutes, 3 ml of 2% Na2CO3 room temperature to thaw. 100 g of fresh mushroom and was added. After vortexing, the mixture was centrifuged at 400 ml solvent (acetone or methanol) were homogenized 250 rpm for 2 hours at room temperature. Absorbance was and stirred overnight. The mixture was incubated for 24 measured at 760 nm. In this study 50-500 µg/ml hours in a 25°C water bath with shaking at 120 rpm. After concentrations of gallic acid were used as the standard. 24 hours the homogenate was filtered through Whatman No. 1 filter paper and the filtrate was stored in the dark. The 2.4.3. Determination of reducing power capacity same process was repeated on the 2nd and 3rd days by Reducing power was analysed using the Oyaizu (1986) adding 300 ml solvent to the remaining mushroom method [16]. 1 ml was taken from each extract sample sediment in Whatman paper. After three days, samples prepared at 100-1000 µg/ml concentrations and mixed with were treated with 1/10 solvent. Supernatants were collected 2.5 ml 0.2 M pH6.6 phosphate buffer and 2.5 ml of 1% and the solvent was evaporated in the evaporator at 35°C. K Fe(CN) . The mixture was incubated for 20 minutes at Obtained extracts were stored at 4°C until use. Although 3 6 50°C. After incubation, 1 ml of sample was taken from the β-glucan content determination was made with fresh mixture and was mixed with 1 ml of distilled water and 0.2 mushroom, antioxidant activity and antimicrobial activity ml of 1% FeCl3, absorbance values were measured at 700 assays made with extracts [11]. nm relative to a reagent blank. 2.3. Determination of β-glucan Content 2.4.4. Guaiacol peroxidase enzyme activity β-glucan content determination of the mushroom species Singh's method was used to determine the guaiacol was made with fresh mushrooms which milled < 1 mm. To peroxidase enzyme activity of mushroom species. determine the β-glucan content a "Mushroom and Yeast According to the method, reaction mixtures were prepared β-glucan K-YBGL 8.4" kit was used [12]. for samples and for controls. (1.25 ml of 50 mM Potassium phosphate buffer pH7.0, 0.5 ml, 1% Guaiacol 0.5 ml, 0.5 2.4. Antioxidant Activity ml 30% Hydrogen peroxide, 0.1 ml filtered mushroom homogenate for enzyme extract) [17]. After preparation of 2.4.1. Determination of DPPH radical scavenging activity reaction mixtures, absorbance of the samples was The free radical scavenging activity of 2,2-diphenyl-1- measured with a UV spectrophotometer at 470 nm every 60 picrylhydrazyl (DPPH) radical was determined using the seconds for 4 minutes (1 Enzyme unit: Amount of enzyme that transforms 1 μmol guaiacol.) (Ɛ=26.6 Mm-1cm-1). Food Science and Technology 6(2): 47-55, 2018 49 2.5. Antimicrobial Activity first diluting 1:50, then 1:20. 10 ml of microorganisms was added to each well. DMSO was added instead of extract as 2.5.1. Test microorganisms a negative control. As a positive control 1 mg/ml ampicillin Test microorganisms used
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