RESUMO
Fungi are an inexhaustible source of bioactive metabolites that can be driven to find medicines for chronic diseases, as Alzheimer's disease. In the present work, five species of soil-originated fungi (Aspergillus chevalieri, Clonostachys rogersoniana, Fusarium nygamai, Penicillium sp., and Talaromyces calidicanius) were submitted to mutual biotic stress aiming at activating the expression of metabolites capable of inhibiting the enzyme acetylcholinesterase. HPLC profiles showed that the in vitro biotic stress triggered the biosynthesis of metabolite-mediated defense responses. Five compounds present in the complex co-culture matrix were identified by Paper Spray Mass Spectrometry (PS-MS). The approach enhanced the biosynthesis of acetylcholinesterase inhibitors (up to 99.6% inhibition) in comparison with the individual cultures. The mutual biotic stress between T. calidicanius and A. Chevalieri led to the biosynthesis of a pool of metabolites statistically as efficient as serine (p < 0.05), the positive control used in the experiments (99.6 and 99.1% inhibition, respectively).
Assuntos
Inibidores da Colinesterase , Penicillium , Acetilcolinesterase/metabolismo , Inibidores da Colinesterase/química , Fungos/metabolismo , Penicillium/metabolismo , Estresse FisiológicoRESUMO
The effect of phenylalanine removal on the peptide profile of the protein hydrolysates from wheat flour wasinvestigated. Nine hydrolysates were prepared, using a successive association of a pancreatin and a crudeenzymatic extract obtained from pineapple peel (CE), and the effect of the order of enzymes addition, thereaction temperature, the enzyme: substrate (E:S) ratio, and the physical treatment of sample were examined.The analysis of peptide profile of hydrolysates was performed in two stages, that is before and after removingphenylalanine. The size-exclusion high performance liquid chromatography was used for performing thefractionation, followed by Correct Fraction Area technique for quantifying peptides and free amino acids.The process of phenylalanine removal improved the peptide profile of three hydrolyzed samples, but it didnot affect the five hydrolysates. The beneficial effect of this process is correlated with the increase of di- andtripeptides contents, or in reducing the amount of large peptides. The best peptide profile was obtainedafter phenylalanine removal by using pancreatin at E:S ratio of 4:100 at 50ºC for 210min, followed by CEat E:S ratio of 10:100 at 70ºC for 90 min.
Para avaliar o efeito da remoção de fenilalanina no perfil peptídico dos hidrolisados proteicos de farinhade trigo, foram preparados nove hidrolisados empregando-se a associação sucessiva de pancreatina e deextrato enzimático bruto obtido da casca de abacaxi (EB). Foram testados o efeito da ordem de adição dasenzimas, da temperatura de reação, da relação enzima: substrato (E:S) e do tratamento físico da amostra.A análise do perfil peptídico dos hidrolisados foi realizada em duas etapas: antes e após a remoção dafenilalanina. A cromatografia líquida de alta eficiência de exclusão molecular foi utilizada para efetuaro fracionamento e a quantificação dos peptídeos e aminoácidos livres pela técnica da Área Corrigida daFração. O processo de remoção de fenilalanina melhorou o perfil peptídico de três hidrolisados, mas nãoafetou no de cinco hidrolisados. O efeito benéfico desse processo está associado ao aumento no teor dedi- e tripeptídeos ou à redução na quantidade de peptídeos grandes. O melhor perfil peptídico foi obtidoapós a remoção de fenilalanina, utilizando-se pancreatina E:S de 4:100 a 50C, durante 210 min, seguidade EB E:S de 10:100 a 70C durante 90 min.