Hemijska karakterizacija i biološka svojstva polisaharidnih ekstrakata gljiva Fomes fomentarius, Auricularia auricula-judae i Sparassis crispa

ector of functional food is more and more appreciated by modern market and consumers. Food is energy, medicine, philosophy, trend and science, now more than ever. Claiming that some type of food has beneficial effect on human health without proving it is past. Active components have to be identified, as well as their biological properties. Furthermore, those components have to be extracted, and the extraction procedure has to be economically justified. Medicinal mushrooms fulfill all listed characteristics, since they contain active compounds like polysaccharides, proteins, triterpene, and phenolic compounds. Polysaccharides isolated from mushrooms are marked as the most complex molecules in nature, which makes them excellent immune response modulators. Biological effects of mushrooms polysaccharides are various due to the differences in their structure, molecular mass, conformation, and water solubility. Although the mechanisms of their action are not completely clear, it was noticed that the ability to dissolve in water strongly affects mushroom’s polysaccharides biological activity. Extraction procedure can have negative impact on biological effect of these molecules, too. The aim of this thesis was to obtain polysaccharide extracts of selected mushroom species, and to modify their structure with enzymes which hydrolyze α-D-glucoside bondages. Then, to characterize extracts, and to examine their biological activity (antimicrobial, antioxidative, cytotoxic and the ability to inhibit angiotensin I-converting enzyme (ACE)). Finally, the extracts were compared, and based on the results of applied methods their chemical characteristics were connected with biological activities. This approach should enable the selection of optimal conditions for the production of biologicaly active mushroom polysaccharides. Overall, results should provide the standardized procedure for the most efficient utilization of mushroom extracts. Polysaccharides of selected mushrooms, Fomes fomentarius, Auricularia auricula-judae and Sparassis crispa, were extracted by hot water and alkali extraction, while crude water extractswere further purified by dialysis...Savremeno tržište i potrošači sve više prepoznaju sektor funkcionalne hrane. Hrana je više nego ikad energija, medicina, filozofija, trend ali i nauka. Tako više nije dovoljno tvrditi da je neka namirnica blagotvorna po zdravlje čoveka. Potrebno je identifikovati aktivne komponente, utvrditi koje biološke osobine ispoljavaju, a potom i na koji način ih je moguće izolovati, te postupak učiniti komercijalnim. Medicinske gljive predstavljaju upravo takvu namirnicu, jer sadrže brojna aktivna jedinjenja poput polisaharida, fenolnih jedinjenja, proteina, triterpena. Polisaharidi gljiva najkompleksniji su molekuli u prirodi i kao takvi modulatori imunog odgovora. Biološki odgovori polisaharida su raznovrsni što je posledica različite građe, molekulske mase, konformacije i rastvorljivosti ovih molekula. Iako mehanizmi delovanja nisu potpuno razjašnjeni uočeno je da biološki efekat polisaharida gljiva može biti umanjen zbog relativno male rastvorljivosti u vodi. Takođe, postupak ekstrakcije može se negativno odraziti na biološka svojstva ekstrakta. Cilj ove teze bilо je dobijanje polisaharidnih ekstrakata odabranih gljiva te njihova strukturna modifikacija primenom specifičnih enzima koji vrše hidrolizu α-D-glikozidnih veza. Zatim, hemijska karakterizacija ekstrakata, a potom ispitivanje bioloških osobina (antimikrobnih, antioksidativnih, citotoksičnosti i sposobnosti inhibicije angiotenzin I-konvertujućeg enzima (ACE)). Konačno, ekstrakti su upoređeni na osnovu rezultata primenjenih metoda, te su njihove hemijske osobine dovedene u vezu sa biološkim svojstvima. To je trebalo da omogući selekciju optimalnih uslova za dobijanje biološki aktivnih polisaharida gljiva, i tako obezbedi standardizacija postupka za njihovo najbolje iskorišćenje. Ekstrakcija polisaharida odabranih gljiva, Fomes fomentarius, Auricularia auricula-judae i Sparassis crispa, izvršena je postupcima vrele vodene i alkalne ekstrakcije, a sirovi vodeni ekstrakt je i naknadno prečišćen dijalizom. Sirovi polisaharidni ekstrakti modifikovani su sukcesivnim dodavanjem izoamilaze, β-amilaze, α-amilaze i pronaze. Hemijska karakterizacija ekstrakata sprovedena je primenom Megazyme glukanskog kita, spektrofotometrijskim određivanjem sadržaja ukupnih ugljenih hidrata, proteina i fenolnih jedinjenja..

Zeolites as possible biofortifiers in maitake cultivation

The levels of Ni, Cu and Mg in Grifola frondosa (also known as Maitake mushroom) fruit body produced on zeolite Minazel Plus (MG)-supplemented substrate were measured with inductively coupled plasma optical emission spectrometry (ICP-OES). Two different concentrations of MG were added to the substrate for mushroom cultivation. Levels of selected metals were measured in cultivated dry carpophores. The content of Ni increased in fruit bodies produced on supplemented substrate, while in case of Cu, a pronounced decrease was observed. When two different concentrations of MG were implemented, the Mg level showed both positive and negative trend, depending on the applied concentration of zeolite. MG in a concentration of 1% showed the strongest influence on the observed elements in the cultivated fruiting body of Maitake mushroom

UTICAJ EKSTRAKATA GLJIVE INONOTUS OBLIQUUS NA FORMIRANJE BIOFILMA PATOGENIH BAKTERIJA

Inonotus obliquus (Čaga) je medicinski značajna glјiva čija micelija je tamne boje, a razvija se u kori breze. Pripada porodici Hymenochaetaceae. Ekstrakti I. obliquus koriste se širom Severne i Istočne Evrope, Koreje i Rusije jer se pretpostavlјa da pokazuju anitimikrobne, antioksidativne, antiinflamatorne i antitumorske aktivnosti. Hemijski testovi pokazuju da I. obliquus proizvodi širok spektar sekundarnih metabolita, uklјučujući fenolna jedinjenja, melanine, uglјene hidrate i proteine. Zadatak ovog istraživanja je bio da se utvrdi hemijski sastav, kao i antimikrobna i antiadhezivna i antibiofilmska sposobnost dobijenih subkritičnih vodenih ekstrakata I. obliquus poreklom iz Mongolije (IM) i sa planine Vlasina, Srbija (IS). Utvrđen je ukupan sadržaj proteina, uglјenih hidrata i fenola, a visok sadržaj fenola u oba ekstrakta. Fenolne kiseline su identifikovane hemijskom analizom HPLC i potvrđeno je prisustvo hlorogenske kiseline u najvećoj koncentraciji, katehina, p-kumarinske kiseline i cimetne kiseline. Antimikrobni potencijal subkritičnih vodenih ekstrakata ispitan je mikrodilucionom metodom. Antiadhezivna i antibiofilmska sposobnost praćena je korišćenjem Gram pozitivnih bakterija Listeria monocitogenes, Enterococcus faecalis i Staphilococcus aureus, kao i Gram negativnih Salmonella enteritidis, Escherichia coli i Pseudomonas aeruginosa. Najznačajnija proizvodnja biofilma zabeležena je kod E. faecalis (treća kategorija) i S. aureus (druga kategorija). Uočeno je da su ekstrakti IS i IM doveli do 90% adhezije kod E. faecalis i S. aureus kao i do redukcije biofilma, odnosno da je došlo do smanje sposobnost ove dve bakterije da formiraju biofilm. Gram pozitivne bakterije su se pokazale kao osetlјivije u prisustvu oba testirana subkritična vodena ekstrakta (MBC - 20 mg/mL u svim eksperimentima). Prema dobijenim vrednostima MIC, IM (5 mg/mL, 2,5 mg/mL, 2,5 mg/mL) je ispolјio jači efekat od IS (10mg/mL, 5 mg/mL, 5 mg/mL) na L. monocitogenes, E. faecalis i S. aureus. Ova istraživanja pokazala su da su testirani subkritični vodeni ekstrakti I. obliquus veoma efikasni u sprečavanju nastanka biofilma, posebno onih odabranih Gram-pozitivnih bakterija. Ovo otkriće bi moglo biti veoma korisno u borbi protiv biofilma, uzimajući u obzir sve probleme i opasnosti koje oni nose

Oyster mushroom (Pleurotus ostreatus) cultivation on different food waste originated substrates

Oyster mushroom (Pleurotus ostreatus) was cultivated on seven substrate mixtures, containing grape pomace, soybean husk and sunflower seed husk, which represent the wastes from the wine and edible oil industry. Wheat straw was used as a control substrate. Grape pomace mixtures and control substrate were grown under higher ambient temperature than soybean and sunflower husk mixtures. The effect of food industry wastebased substrates on several productivity parameters (time required for completion of mycelium running (days), mycelium running rate in bags (mm/days), time required for primordia initiation (days), time required for harvesting (days), total mushroom yield and biological efficiency) was estimated. Results indicated soybean husk affecting the best mushroom yield and biological efficiency, followed by sunflower seed husk substrates and wheat straw as a control substrate. Grape pomace-based substrates showed the lowest productivity potential. On the other hand, lower cultivation temperatures showed slower growth causing the longer time from inoculation to harvest. To complete the estimation of these alternative raw materials for mushroom cultivation future research may include the assessment of the chemical and antioxidant properties of mushrooms as well as their quality characteristics

Uticaj ekstrakata gljive Inonotus obliquus na formiranje biofilma patogenih bakterija

Inonotus obliquus (Čaga) je medicinski značajna glјiva čija micelija je tamne boje, a razvija se u kori breze. Pripada porodici Hymenochaetaceae. Ekstrakti I. obliquus koriste se širom Severne i Istočne Evrope, Koreje i Rusije jer se pretpostavlјa da pokazuju anitimikrobne, antioksidativne, antiinflamatorne i antitumorske aktivnosti. Hemijski testovi pokazuju da I. obliquus proizvodi širok spektar sekundarnih metabolita, uklјučujući fenolna jedinjenja, melanine, uglјene hidrate i proteine. Zadatak ovog istraživanja je bio da se utvrdi hemijski sastav, kao i antimikrobna i antiadhezivna i antibiofilmska sposobnost dobijenih subkritičnih vodenih ekstrakata I. obliquus poreklom iz Mongolije (IM) i sa planine Vlasina, Srbija (IS). Utvrđen je ukupan sadržaj proteina, uglјenih hidrata i fenola, a visok sadržaj fenola u oba ekstrakta. Fenolne kiseline su identifikovane hemijskom analizom HPLC i potvrđeno je prisustvo hlorogenske kiseline u najvećoj koncentraciji, katehina, p-kumarinske kiseline i cimetne kiseline. Antimikrobni potencijal subkritičnih vodenih ekstrakata ispitan je mikrodilucionom metodom. Antiadhezivna i antibiofilmska sposobnost praćena je korišćenjem Gram pozitivnih bakterija Listeria monocitogenes, Enterococcus faecalis i Staphilococcus aureus, kao i Gram negativnih Salmonella enteritidis, Escherichia coli i Pseudomonas aeruginosa. Najznačajnija proizvodnja biofilma zabeležena je kod E. faecalis (treća kategorija) i S. aureus (druga kategorija). Uočeno je da su ekstrakti IS i IM doveli do 90% adhezije kod E. faecalis i S. aureus kao i do redukcije biofilma, odnosno da je došlo do smanje sposobnost ove dve bakterije da formiraju biofilm. Gram pozitivne bakterije su se pokazale kao osetlјivije u prisustvu oba testirana subkritična vodena ekstrakta (MBC - 20 mg/mL u svim eksperimentima). Prema dobijenim vrednostima MIC, IM (5 mg/mL, 2,5 mg/mL, 2,5 mg/mL) je ispolјio jači efekat od IS (10mg/mL, 5 mg/mL, 5 mg/mL) na L. monocitogenes, E. faecalis i S. aureus. Ova istraživanja pokazala su da su testirani subkritični vodeni ekstrakti I. obliquus veoma efikasni u sprečavanju nastanka biofilma, posebno onih odabranih Gram-pozitivnih bakterija. Ovo otkriće bi moglo biti veoma korisno u borbi protiv biofilma, uzimajući u obzir sve probleme i opasnosti koje oni nose

An insight into in vitro antioxidant activity of Cantharellus cibarius hot water extract for the potential application in meat products

The current research was undertaken to estimate the in vitro antioxidant activity of Cantharellus cibarius mushroom extracted by boiling in water for 30 minutes. Several previous studies have shown that the addition of edible mushrooms in meat products affects the reduction of lipid oxidation and prolongs the shelf-life of the final products. Antioxidant capacity of C. cibarius was measured using the following methods: reducing power ability, lipid peroxidation assay, cupric ion reducing antioxidant capacity (CUPRAC) and DPPH free radical scavenging activity. Reducing power and antioxidant activity assays of C. cibarius hot water extract showed low antioxidant activity. CUPRAC assay demonstrated positive effect only at the concentration of 10 mg/mL, whereas DPPH radical scavenging activity showed moderate antioxidant activity in comparison with culinary-medicinal mushrooms, with the effective concentration (EC50) from 7.41 mg/mL

Nutraceutical properties of the methanolic extract of edible mushroom Cantharellus cibarius (Fries): primary mechanisms

The methanolic extract of the wild edible mushroom Cantharellus cibarius Fr. (chanterelle) was analyzed for in vitro antioxidative, cytotoxic, antihypertensive and antibacterial activities. Various primary and secondary metabolites were found. Phenols were the major antioxidant components found in the extract (49.8 mg g(-1)), followed by flavonoids, whose content was approximately 86% of the total phenol content. Antioxidant activity, measured by four different methods, was high for inhibition of lipid peroxidation (EC50 = 1.21 mg mL(-1)) and chelating ability (EC50 = 0.64 mg mL(-1)). The antioxidant activity of the C. cibarius methanol extract was achieved through chelating iron compared to hydrogen atom and/or electron transfer. The extract showed good selectivity in cytotoxicity on human cervix adenocarcinoma HeLa, breast carcinoma MDA-MB-453 and human myelogenous leukemia K562, compared to normal control human fetal lung fibroblasts MRC-5 and human lung bronchial epithelial cells BEAS-2B. The extract had inhibitory activity against angiotensin converting I enzyme (ACE) (IC50 = 0.063 mg mL(-1)). The extract revealed selective antimicrobial activity against Gram-positive bacteria with the highest potential against E. faecalis. The medicinal and health benefits, observed in wild C. cibarius mushroom, seem an additional reason for its traditional use as a popular delicacy food

Antioxidant properties of Subcritical Water Extracts Derived from Mushroom Inonotus obliquus

The medicinal mushroom Inonotus obliquus is widespread in Europe, Asia and North America. In many countries, since the 16 th century, it has traditionally been used to treat gastrointestinal cancer, cardiovascular disease and diabetes [1]. This study was designed to determine the chemical composition and antioxidant activity of subcritical water extracts obtained from fruiting bodies of I. obliquus originating from Mongolia (IM) and from the mountain Vlasina, Serbia (IS). Chemical analysis revealed the total content of proteins, carbohydrates and phenols. High carbohydrate content was found in both extracts, and glucose was the most dominant monosaccharide. In order to identify phenolic acids, the extracts were subjected to a qualitative chemical analysis, and the presence of chlorogenic acid, catechin, p-coumaric acid and cinnamic acid has been confirmed. Chlorogenic acid was detected in the highest concentration, compared to other phenolic acids. DPPH free radical scavenging activity assay was used to measure the antioxidant properties of extracts in vitro . Extracts concentrations from 0.156 to 10 mg/mL were tested and a maximum of 93% of scavenging ability was reached. The results indicated that antioxidant activity in both extracts can be achieved through hydrogen atom (HAT) and single electron transfer (SET) as dominant mechanisms [2]

Artificial Neural Network Prediction of Antiadhesion and Antibiofilm-Forming Effects of Antimicrobial Active Mushroom Extracts on Food-Borne Pathogens

The problem of microbial biofilms has come to the fore alongside food, pharmaceutical, and healthcare industrialization. The development of new antibiofilm products has become urgent, but it includes bioprospecting and is time and money-consuming. Contemporary efforts are directed at the pursuit of effective compounds of natural origin, also known as “green” agents. Mushrooms appear to be a possible new source of antibiofilm compounds, as has been demonstrated recently. The existing modeling methods are directed toward predicting bacterial biofilm formation, not in the presence of antibiofilm materials. Moreover, the modeling is almost exclusively targeted at biofilms in healthcare, while modeling related to the food industry remains under-researched. The present study applied an Artificial Neural Network (ANN) model to analyze the anti-adhesion and anti-biofilm-forming effects of 40 extracts from 20 mushroom species against two very important food-borne bacterial species for food and food-related industries—Listeria monocytogenes and Salmonella enteritidis. The models developed in this study exhibited high prediction quality, as indicated by high r2 values during the training cycle. The best fit between the modeled and measured values was observed for the inhibition of adhesion. This study provides a valuable contribution to the field, supporting industrial settings during the initial stage of biofilm formation, when these communities are the most vulnerable, and promoting innovative and improved safety management. © 2023 by the authors

The effect of green extraction method, subcritical, water extraction, on selected biological activities of Chaga mushroom

Inonotus obliquus, or Chaga, is a medicinal mushroom with a long history of folk medicinal use and growing popularity worldwide. It cannot be cultivated so novel, greener extraction methods are needed to satisfy the market and obtain high-quality supplements. In this study, we evaluated the biological activity (antimicrobial and antioxidant) of Mongolian (IM) and Serbian (IS) Chaga extracts obtained by subcritical water extraction. They were also screened for general chemical composition. Both IM and IS, have high phenolic content. HPLC analysis revealed that extracts are especially rich in chlorogenic acid, followed by catechins, p-coumaric and cinnamic acids. ABTS assay confirmed the remarkable scavenging ability of extracts, reaching 98% when a concentration of 2 mg/mL was tested. The antimicrobial potential was examined by microdilution method using Gram-positive and (Enterococcus faecalis and Staphylococcus aureus), Gram-negative bacteria (Escherichia coli and Actinetobacteria baumannii). According to the obtained minimal inhibitory concentration (MIC) for E. faecalis S. aureus (MIC -1.25 mg / mL), E. coli, and A. baumannii (MIC -2.5 mg / mL) both extracts were shown to have the same effect on the tested bacteria. Hereby, we demonstrated the high biological potential of extracts obtained by subcritical water extraction