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Evelyn de Souza Oliveira Lopeshttp://lattes.cnpq.br/1693469786109191Inayara Cristina Alves LacerdaRaquel Linhares Bello de AraújoDenise Sande Santoshttp://lattes.cnpq.br/6935148677614355Verônica de Souza Miranda Pacheco2023-04-17T14:26:20Z2023-04-17T14:26:20Z2016-06-30http://hdl.handle.net/1843/52065As celulases correspondem às enzimas com a terceira maior aplicação industrial do mundo. São amplamenteutilizadas nas indústrias têxtil, de papel e celulose, extração de sucos, detergentes, nutrição animal e bioetanol. A produção de enzimas utilizando microrganismos é preferencial devido à diversidade bioquímica e facilidade de manipulação celular. Os fungos filamentosos possuem grande potencial para secretar enzimas e são entre os microrganismos os maiores produtores de celulases. O objetivo desse estudo foi isolar e selecionar fungos filamentosos, de resíduos agroindustriais, com potencial atividade celulolítica. Os fungos filamentosos foram isolados dos resíduos agroindustriais: bagaço de cana de açúcar, casca de amendoim e casca de coco seco. Foram isolados 28 fungos filamentosos desses resíduos e selecionados sete potencialmente produtores de enzimas celulolíticas pertencentes aos gêneros Aspergillus sp. Penicillium sp. Curvalaria sp. Os ensaios qualitativos de produção de celulase foram realizados em placas de Petri contendo ágar carboximetilcelulose (CMC) e foi calculado o Índice enzimático (IE). Os sete fungos filamentosos que apresentaram resultado positivo no ensaio qualitativo foram cultivados em em meio líquido contendo celulose microcristalina. No caldo fermentado (preparado bruto enzimático) foi avaliada a produção de celulases pelo teste de difusão em placas com gel e determinada a atividade celulolítica total FPase (Filter Paper Cellulase). Os fungos filamentosos que formaram halos claros, no ensaio qualitativo de difusão em gel em placa de Petri, foram submetidos ao microcultivo para a identificação convencional dos gêneros. Os IE variaram entre 1,92 e 8,54. Todos os fungos filamentosos que apresentaram IE maior do que 1 (um) apresentaram atividade celulolítica total no ensaio quantitativo. Os valores de FPase variaram entre 0,01299 FPU.mL-1 (Penicillium sp. BM9 com 48h de fermentação) e 0,05013 FPU.mL -1 (Aspergillus sp. CO2 com 72h de fermentação) e os valores de atividade específica variaram entre 0,04755 FPU.mg-1 de proteína total (Aspergillus sp. CO2 com 24h de fermentação) e 0,56407 FPU.mg-1 de proteína total (BM18 com 72h de fermentação). Estes resultados confrontados com a literatura mostraram-se promissores. Existe potencial para obtenção com esses fungos, de atividade celulolítica superior aos valores encontrados, para isso são necessários estudos de otimização da produção. Estudos futuros investigando a a atividade de endoglucanases e βglicosidases também são necessários para avaliar o sinergismo entre essas enzimas, já que os fungos produzem diferentes quantidades de enzimas do complexo celulolítico.Cellulases are the third largest industrial enzymes worldwide. These enzymes are applied in various industries such as textile, pulp and paper, juice extraction, detergents, animal feed and bioethanol production. Enzymes produced by microorganisms are preferred due to their biochemical diversity and ease of genetic manipulation. Filamentous fungi have shown their great potential for secreting enzymes. The present work aimed to isolate and select filamentous fungi from agroindustrial residues with potential cellulolytic activity. The filamentous fungi were isolated from the following agroindustrial residues: sugarcane bagasse, peanut shell and coconut shell, and selected in Petri dishes containing carboxymethylcellulose (CMC) agar according to their Enzymatic Index (EI). The obtainment of crude enzymatic preparation to quantify the total cellulolytic activity (FPase) occurred under submerged fermentation by filamentous fungi. The FPase quantitative assay was performed using 3.5 dinitroxalicylic acid on a microplate reader with absorbance at 540 nm. The specific activity of crude enzymatic preparations was also quantified. The filamentous fungi that formed light halos around the wells in the gel diffusion assay were submitted to the microculture for conventional identification of genera. Twenty-eight filamentous fungi were isolated. Among these, seven filamentous fungi potentially producers of cellulolytic enzymes, which belonged to the genus Aspergillus sp., Penicillium sp. and Curvalaria sp., were selected. EI ranged from 1.92 to 8.54. All filamentous fungi that presented EI greater than 1 (one) produced crude enzymatic preparation with total cellulolytic activity also evidenced in the qualitative gel diffusion assay. PFase values varied between 0.01299 filter paper units per mililiter (FPU.mL-1 (Penicillium sp. BM9 with 48 h of fermentation) and 0.05013 FPU.mL-1 (Aspergillus sp. CO2 with 72 h of fermentation) and the values of specific activity ranged from 0.04755 FPU.mg-1 of protein (Aspergillus sp. CO2 with 24 h fermentation) and 0.56407 FPU.mg-1 protein (BM18 with 72 h of fermentation). These results were promising when compared with the results of literature. There is potential for the obtainment of values of cellulolytic activity higher than the values found in the present study, so further studies of production optimization are required. Future studies investigating endoglucanases and β-glycosidases are needed to evaluate synergism because fungi produce different amounts of enzymes of the cellulolytic complex.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Ciência de AlimentosUFMGBrasilFARMACIA - FACULDADE DE FARMACIAhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessCelulasesSeleção de fungos filamentososAtividade celulolítica totalFPaseIsolamento e seleção de fungos filamentosos produtores de celulases a partir de resíduos agroindustriais.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALDESSERTAÇÃO VERÔNICA DE SOUZA MIRANDA PACHECO 2016.pdfDESSERTAÇÃO VERÔNICA DE SOUZA MIRANDA PACHECO 2016.pdfapplication/pdf1905753https://repositorio.ufmg.br/bitstream/1843/52065/1/DESSERTA%c3%87%c3%83O%20VER%c3%94NICA%20DE%20SOUZA%20MIRANDA%20PACHECO%20%202016.pdf70e9634d547ebb5701e59f31eb141718MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/52065/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/52065/3/license.txtcda590c95a0b51b4d15f60c9642ca272MD531843/520652023-04-17 11:26:21.158oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-04-17T14:26:21Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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