Bioprospection of Secondary Metabolites in Isolates of Shiitake Mushroom (Lentinula edodes (Berk.) Pegler)

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2022-08-12T13:56:14Z No. of bitstreams: 1 2019 - Rafaela de Souza Menezes.pdf: 653046 bytes, checksum: f0184a7056a21faaa6c2678d0f0fd7f1 (MD5) Made available in DSpace on 2022-08-12T13:56:14Z (GMT). No. of bitstreams: 1 2019 - Rafaela de Souza Menezes.pdf: 653046 bytes, checksum: f0184a7056a21faaa6c2678d0f0fd7f1 (MD5) Previous issue date: 2019-08-29 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior The upward demand and acceptance, the mushroom Lentinula edodes (Berk.) Pegler (shiitake mushroom) is among the most consumed species in Brazil. The need for studies that investigate its morphological, genetic characterization and bioactive composition present in the species are increasing as well. Because of this, some studies related to its cultivation, medicinal and nutraceutical values are becoming more frequent. Genetically, transcribed internal spacer regions (ITS) have been widely used to define interspecific variations, including for members of Fungi kingdom. For metabolic characterization, chromatographic techniques are used to bioprospected actives with biotechnological potential. Thus, this research sought to sequence ITS regions in order to promote the confirmation of species classification of the target isolates of the study, as well as consider the survey of secondary metabolites contained in mycelium samples from these isolates. Approximately 625 bp size amplification products were obtained. The isolates of the present study grouped with the L. edodes isolates from the database and differed from the other species representing the genus Lentinula. Sequence analysis of the ITS region was adequate for interspecific differentiation of the genus Lentinula. Confirming its potential for identification and molecular characterization of fungi. These samples were also followed for quantitative analysis, in which the mobile phase components of these samples were identified and quantified in High Performance Liquid Chromatography-Diodes Arrangement Detector (HPLC-DAD). In addition, spectrophotometer analysis of total antioxidant and phenolic compounds was performed. Regarding total phenolic compounds, a concentration of 0.48 mg gallic acid / ml mycelium extract was observed. The chemical fractionation performed with mycelium showed that these secondary metabolites can be observed by the liquid chromatography technique, with the presence of five possible biocompounds with different yield times in the generated chromatogram. Through the injection of patterns, the same times were observed for the structure of 2,4 dimethoxybenzoic acid and 2,4 dihydroxybenzoic acid. Thus, the present work found values that justify the metanolic extraction of these metabolites also demonstrates low which reinforced that the nature of these compounds in mushrooms are polar Com a crescente comercializa??o e aceita??o, o cogumelo Lentinula edodes est? entre as esp?cies mais consumidas no Brasil, surgindo a necessidade de sua caracteriza??o morfol?gica, gen?tica e com rela??o ? composi??o de bioativos presentes na esp?cie. Por conta disso alguns estudos relacionados ao seu cultivo, valor medicinal e nutrac?utico s?o cada vez mais frequentes. No ?mbito gen?tico, as regi?es espa?adoras internas transcritas (ITS) t?m sido amplamente utilizadas para definir varia??es interespec?ficas, inclusive para integrantes do reino Fungi. J? para caracteriza??o metab?lica s?o lan?adas m?o de t?cnicas cromatogr?ficas para o levantamento de bioativos com potencial biotecnol?gico. Com isso, o presente trabalho procurou sequenciar regi?es ITS com o intuito de promover a confirma??o da classifica??o da esp?cie dos isolados alvo da pesquisa, assim como considerar o levantamento de metab?litos secund?rios contidos em amostras de mic?lio destes mesmos isolados. Foram obtidos produtos de amplifica??o de tamanho de aproximadamente 625 pb. Os isolados do presente trabalho agruparam com os isolados de L. edodes provenientes do banco de dados e diferenciaram-se das demais esp?cies representantes do g?nero Lentinula. A an?lise de sequ?ncias da regi?o ITS foi adequada para diferencia??o interespec?fica do g?nero Lentinula, confirmando seu potencial para identifica??o e caracteriza??o molecular de fungos. Essas amostras seguiram para an?lises quantitativas, em que os componentes da fase m?vel destas amostras foram identificados e quantificados em Cromatrografia L?quida de Alta Efici?ncia- Detector de Arranjo de Diodos (CLAE- DAD). Al?m disso, foi feita an?lise por espectrofot?metro de compostos antioxidantes e fen?licos totais. Com rela??o aos compostos fen?licos totais, foi obeservado uma concentra??o de 0,48 mg de ?cido g?lico/ml de extrato de mic?lio. O fracionamento qu?mico realizado com mic?lio, evidenciou que estes metab?litos secund?rios s?o pass?veis de observa??o atrav?s da t?cnica de cromatagrofia l?quida, com isso foram observados no cromatograma gerado, a presen?a de cinco poss?veis biocompostos com diferentes tempos de renten??o. Atrav?s da inje??o de padr?es foram observados os mesmos tempos para estrutura do ?cido 2,4 dimetoxibenz?ico e 2,4 dihidroxibenz?ico. Com isso, o presente trabalho encontrou valores que justificam que a extra??o alco?lica destes metab?litos tamb?m se demonstra baixa refor?ando que a natureza destes compostos em cogumelos s?o de natureza polar