Biological activity and chemical caracteristics of selected extracts of autochtonous macrofungi

Abstract

<p>Prema postavljenim ciljevima u ovoj doktorskoj disertaciji sakupljeno je i&nbsp;<br />determinisano&nbsp;ukupno&nbsp;sedam&nbsp;vrsta&nbsp;autohtonih&nbsp;gljiva&nbsp;sa&nbsp;područja&nbsp;Fru&scaron;ke&nbsp;gore, Tare i&nbsp; Vr&scaron;ačkog brega, pet lignikolnih&nbsp;‐&nbsp;<em>Bjerkandera adusta</em>,&nbsp;<em>Pleurocybella&nbsp; porrigens</em>,&nbsp;St<em>ereum&nbsp;hirsutum,&nbsp;Stereum&nbsp;subtomentosum</em>&nbsp;i&nbsp;Trametes&nbsp;versicolor,&nbsp;i&nbsp;<br />dve&nbsp; terikolne&nbsp;‐&nbsp;<em>Amanita&nbsp; strobiliformis</em>&nbsp;i Hydnum repandum. Utvrđena je&nbsp; njihova&nbsp;biolo&scaron;ka&nbsp;aktivnost&nbsp;(antiradikalska,&nbsp;antioksidativna,&nbsp;antimikrobna,&nbsp;anti‐acetilholinesterazna&nbsp;i&nbsp;citotokisčna)&nbsp;spram&nbsp;hemijskog&nbsp;sastava&nbsp;njihovih&nbsp;vodenih&nbsp;<br />(H₂O),&nbsp;etanolnih&nbsp;(EtOH),&nbsp;metanolnih&nbsp;(MeOH)&nbsp;i&nbsp;polisaharidnih&nbsp;(PSH)&nbsp;ekstrakata.&nbsp;<br />Analiza&nbsp;hemijskog&nbsp;sastava&nbsp;odabranih&nbsp;vrsta&nbsp;uključila&nbsp;je&nbsp;određivanje&nbsp;hemijske&nbsp;<br />karakterizacije&nbsp;PSH&nbsp;ekstrakata&nbsp;‐&nbsp;FTIR&nbsp;analizom,&nbsp;određivanje&nbsp;fenolnog&nbsp;profila&nbsp;‐&nbsp;<br />HPMC/MS‐MS,&nbsp;sadržaja&nbsp;organskih&nbsp;kiselina&nbsp;‐&nbsp;HPLC,&nbsp;sadržaja&nbsp;masnih&nbsp;kiselina&nbsp;‐&nbsp;<br />GC‐FID&nbsp;i&nbsp;sadržaja&nbsp;biogenih&nbsp;elemenata&nbsp;‐&nbsp;AAS.&nbsp;Spektrofotometrijskim&nbsp;metodama<br />određen je ukupan sadržaj proteina i ukupan sadržaj fenola i flavonoida.&nbsp;<br />Prema&nbsp;antiradikalskoj&nbsp;aktivnosti&nbsp;OH^{&bull; ,&nbsp;}O2^&bull;‐,&nbsp;OH^&bull;,&nbsp;Asc^&bull;,&nbsp;DPPH^&bull;&nbsp; i&nbsp;ABTS^&bull;+&nbsp;izdvojili&nbsp;<br />su&nbsp; se&nbsp; ekstrakti lignikolnih vrsta:&nbsp; MeOH ekstrakt vrste&nbsp;<em>P. porrigens</em>, H₂O&nbsp; ekstrakt&nbsp;<em>P.&nbsp;porrigens</em>,&nbsp;MeOH&nbsp;ekstrakt&nbsp;<em>T.&nbsp;versicolor</em>,&nbsp;H₂O&nbsp;ekstrakt&nbsp;<em>S.&nbsp;hirsutum, </em>MeOH&nbsp;ekstrakt&nbsp;<em>S.&nbsp;subtomentosum</em>&nbsp;i&nbsp;H₂O&nbsp;ekstrakt&nbsp;<em>B.&nbsp;adusta</em>,&nbsp;navedenim&nbsp;redom.&nbsp;<br />Najjaču antioksidativnu aktivnost dobijenu FRAP i polarografskom HPMC&nbsp;<br />metodom ispoljili su PSH i H₂O ekstrakti terikolne vrste&nbsp;<em>A. strobiliformis</em>.&nbsp;<br />Antimikrobna aktivnost analiziranih ekstrakata određena je ispitivanjem&nbsp;<br />antibakterijskog,&nbsp;antifungalnog&nbsp;i&nbsp;antiviralnog&nbsp;potencijala,&nbsp;pri&nbsp;čemu&nbsp;se&nbsp;izdvojila&nbsp;<br />vrsta&nbsp;<em> H.&nbsp; repandum</em>&nbsp;ispoljavajući najbolji efekat na Gram‐pozitivne i Gram‐<br />negativne bakterije i na sve analizirane fitopatogene izolate&nbsp;<em>(Fusarium&nbsp;</em>i&nbsp;<br />Alternaria)&nbsp;i<em>&nbsp;T.&nbsp;versicolor</em>&nbsp;na&nbsp;analizirani&nbsp;bakteriofag.&nbsp;Anti‐acetilholinesterazna&nbsp;<br />aktivnost određena je testovima in solid i in liquid, a najbolji procenat&nbsp;<br />inhibicije AChE ispoljili su EtOH ekstrakti vrsta&nbsp;<em>S. hirsutum</em>,&nbsp;<em>B. adusta</em>,&nbsp;<em>S</em>.&nbsp;<br /><em>subtomentosum</em>&nbsp;i&nbsp;<em>T.&nbsp;versicolor</em>.&nbsp;Citotoksična&nbsp;aktivnost&nbsp;ekstrakata&nbsp;određena&nbsp;je&nbsp;<br />MTT testom, a prema najboljoj ispoljenoj aktivnosti izdvojili su se MeOH&nbsp;<br />ekstrakt<em>&nbsp;P.&nbsp;porrigens&nbsp;</em>i&nbsp;ekstrakti<em>&nbsp;B.&nbsp;adusta</em>,&nbsp;H₂O&nbsp;i&nbsp;EtOH.&nbsp;Citotoksična&nbsp;aktivnost&nbsp;<br />ovih&nbsp;lignikolnih&nbsp;vrsta&nbsp;naročito&nbsp;je&nbsp;izražena&nbsp;nakon&nbsp;72&nbsp;h.&nbsp;Na&nbsp;osnovu&nbsp;dobijenih&nbsp;<br />rezultata, gde su se istakle različite vrste i njihovi različiti ekstrakti u&nbsp;<br />primenjenim&nbsp;testovima,&nbsp;jasno&nbsp;je&nbsp;da&nbsp;biolo&scaron;ka&nbsp;aktivnost&nbsp;i&nbsp;hemijski&nbsp;sastav&nbsp;zavise&nbsp;<br />od&nbsp;porekla,&nbsp;vrste&nbsp;i&nbsp;tipa&nbsp;ekstrakta&nbsp;analiziranih&nbsp;gljiva.&nbsp;Na&nbsp;osnovu&nbsp;tipa&nbsp;rastvarača&nbsp;<br />odnosno ekstrakata vrsta, koje su pokazale najbolju aktivnost spram pomenutih&nbsp; testova i na osnovu dobijenih korelacija kao i na osnovu detektovanih jedinjenja,&nbsp; pretpostavljamo da su za ispoljene aktivnosti u najvećoj meri odgovorna fenolna&nbsp; jedinjenja&nbsp;i&nbsp;polisaharidi.&nbsp;&nbsp;<br />&nbsp;<br />Ispoljeni&nbsp;biopotencijal&nbsp;analiziranih&nbsp;vrsta&nbsp;gljiva&nbsp;upućuje&nbsp;na&nbsp;njihovu&nbsp;potencijalnu&nbsp;<br />upotrebu kao funkcionalne hrane i nutraceutika, kao i u biokontroli&nbsp;<br />fitopatogena.</p>

<p>According&nbsp; to&nbsp; the&nbsp; set&nbsp; aims&nbsp; of&nbsp; the&nbsp; presented&nbsp; PhD&nbsp; thesis,&nbsp; seven&nbsp; autochthonous fungal species from the region of Fruska Gora, Tara&nbsp; and Vr&scaron;ac Mountains were collected and determined: five lignicolous ‐ <em>Bjerkandera&nbsp; adusta,&nbsp; Pleurocybella&nbsp; porrigens,&nbsp; Stereum&nbsp; hirsutum,&nbsp; Stereum subtomentosum and Trametes versicolor</em>, and two terricolous ‐ <em>Amanita strobiliformis </em>and <em>Hydnum repandum</em>. Biological activity of these&nbsp; species&nbsp; (antiradical,&nbsp; antioxidant,&nbsp; antimicrobial,&nbsp; anti‐ acetylcholinesterase and cytotoxic) was determined in relation to the chemical composition of the extracts, aqueous (H₂O), ethanolic (EtOH), methanolic (MeOH) and polysaccharide (PSH). Analysis of the chemical content of analyzed species included chemical characterization of PSH extracts&nbsp; &ndash;&nbsp; by&nbsp; FTIR&nbsp; analysis,&nbsp; determination&nbsp; of&nbsp; phenolic&nbsp; profile ‐ by HPMC/MS‐MS, content of organic acids ‐ by HPLC, fatty acid content ‐ by&nbsp; GC‐FID&nbsp; and&nbsp; content&nbsp; of&nbsp; biogenic&nbsp; elements ‐ by&nbsp; AAS. Spectrophotometric methods were applied for determination of the content of total proteins, polyphenols and flavonoids. According to the antiradical activity obtained towards OH^&bull;, О2^&bull;‐, OH^&bull;, Asc^&bull;, DPPH^&bull;&nbsp;and ABTS^&bull;+ extracts of lignicolous species were singled out: <em>P. porrigens&nbsp;</em> (MeOH&nbsp; extract),&nbsp;<em> P.&nbsp; porrigens</em>&nbsp; (H₂O&nbsp; extract),&nbsp;<em>T. versicolor</em>&nbsp;(MeOH&nbsp;&nbsp; extract),<em> S. hirsutum </em>(H₂O extract), <em>S. subtomentosum</em> (MeOH extract) and&nbsp;<em>B. austa</em> (H₂O&nbsp; extract),&nbsp; respectively. The&nbsp; highest antioxidant activity obtained by FRAP and the polarographic HPMC method was exhibited&nbsp; for PSH&nbsp; and&nbsp; H₂O extracts of the terricolous species&nbsp;<em>A. strobiliformis</em>. The intimicrobial activity of analyzed extracts was determined by examination of antibacterial, antifungal and antiviral potentials, whereby&nbsp; the species&nbsp; <em>H.&nbsp; repandum&nbsp;</em>was separated by exhibiting the best effect on Gram‐positive&nbsp; and Gram‐negative bacteria, and all the analyzed hytopathogenic isolates (<em>Fusarium, Alternaria</em>)&nbsp; and <em>T. versicolor</em>&nbsp;against analyzed bacteriophage. Anti‐cetylcholinesterase activity was determined by tests in solid and in liquid, while the best&nbsp; percent of AChE inhibition was showed by EtOH extracts of the species <em>S. hirsutum, B. adusta, S. subtomentosum </em>and <em>T. versicolor</em>.bThe cytotoxic activity of extracts was determined by MTT assay, and according to the best activity, the MeOH extract of <em>P. porrigens</em>, and H₂O and EtOH extracts of <em>B. adusta</em>&nbsp;were distinguished particularly after 72 h. Based on the results obtained, favoring different species and their different extracts in the applied tests, it is clear that the biological activity and chemical composition depend on the origin, species and type of extract of the analyzed fungi. Based on the type of solvent or extract of the species that showed the best activity in relation to the above tests and on the basis of the obtained correlations as well as on&nbsp; the basis of the detected compounds, we assume&nbsp; that&nbsp; the&nbsp; phenol compounds&nbsp; and&nbsp; polysaccharides&nbsp; are responsible for the activities performed.<br />Demonstrated bio‐potential of analyzed fungal species indicates their&nbsp; potential use as functional foods and nutraceutics, as well as in the biocontrol of phytopathogens.</p>