Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae)
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/6444 |
Resumo: | In the present work it was developed analytical methodologies for investigation of biosynthesis of bis-phenylpropanoyl amides, using labeled precursors in the culture medium. The effect of different nitrogen sources on the bis-phenylpropanoyl amides production was also evaluated. These amides are reported as antiparasitary compounds and they also showed activity against gram-positive Bacillus subtilis in bioassays conducted at LaBioMMi (DQ/UFSCar). The analytical methodologies developed were based on High Performance Liquid Chromatography (HPLC) and Mass Spectrometry techniques, mainly electrospray ionization (ESI) and MS/MS. Using the methodologies developed for preparative liquid chromatography it were isolated the bis-phenylpropanoyl amides, brasiliamide A and B and other secondary metabolites co-produced. Liquid chromatography coupled with mass spectrometry techniques allowed fragmentation studies which lead to selective detection experiments of brasiliamides in mediums enhanced with different nitrogen sources. The quantitative analysis (HPLC/UV-MS/MS) indicated great influence of the amino acid L-phenylalanine on the bis-phenylpropanoyl amides production. From these results, the biosynthesis of the phenylapropanoids was investigated using [2-13CPhe]. The RMN 13C and HPLC/UV-MS analysis showed the incorporation of two amino acid units on brasiliamides structure formed by the phenylpropanoids pathway, which is uncommon in fungi. The first phenylpropanoid pathway step was evaluated throw using enzymatic extract with Phenylalanine ammoniolyase (PAL). The HPLC/UV analysis indicates the formation of cinnamic acid from Lphenylalanine. |
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Fill, Taícia PachecoRodrigues Filho, Edsonhttp://lattes.cnpq.br/3667941735597178http://lattes.cnpq.br/07416383797071548e5be67e-894c-48a2-94a3-701e47160b772016-06-02T20:36:21Z2009-10-302016-06-02T20:36:21Z2009-02-12FILL, Taícia Pacheco. Biosynthetic study of phenylpropanoyl amides produced by The fungus Penicillium brasilianum, an endophyte from Melia azedarach. 2009. 183 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2009.https://repositorio.ufscar.br/handle/ufscar/6444In the present work it was developed analytical methodologies for investigation of biosynthesis of bis-phenylpropanoyl amides, using labeled precursors in the culture medium. The effect of different nitrogen sources on the bis-phenylpropanoyl amides production was also evaluated. These amides are reported as antiparasitary compounds and they also showed activity against gram-positive Bacillus subtilis in bioassays conducted at LaBioMMi (DQ/UFSCar). The analytical methodologies developed were based on High Performance Liquid Chromatography (HPLC) and Mass Spectrometry techniques, mainly electrospray ionization (ESI) and MS/MS. Using the methodologies developed for preparative liquid chromatography it were isolated the bis-phenylpropanoyl amides, brasiliamide A and B and other secondary metabolites co-produced. Liquid chromatography coupled with mass spectrometry techniques allowed fragmentation studies which lead to selective detection experiments of brasiliamides in mediums enhanced with different nitrogen sources. The quantitative analysis (HPLC/UV-MS/MS) indicated great influence of the amino acid L-phenylalanine on the bis-phenylpropanoyl amides production. From these results, the biosynthesis of the phenylapropanoids was investigated using [2-13CPhe]. The RMN 13C and HPLC/UV-MS analysis showed the incorporation of two amino acid units on brasiliamides structure formed by the phenylpropanoids pathway, which is uncommon in fungi. The first phenylpropanoid pathway step was evaluated throw using enzymatic extract with Phenylalanine ammoniolyase (PAL). The HPLC/UV analysis indicates the formation of cinnamic acid from Lphenylalanine.ESTUDOS BIOSSINTÉTICOS DE AMIDAS BIS-FENILPROPANOÍDICAS PRODUZIDAS PELO FUNGO Penicillium brasilianum, UM ENDOFÍTICO DE Melia azedarach. Foi o título do trabalho no qual foram desenvolvidas metodologias de análise para a investigação da biossíntese de amidas bisfenilpropanoídicas, pela adição de precursores isotopicamente marcados ao meio de cultura e otimização da produção destas amidas pelo micro-organismo quando submetido à variadas fontes nitrogenadas. As amidas, substâncias relatadas como bons antiparasitários, apresentaram também atividade contra a bactéria gram-positiva Bacillus subtilis. As metodologias de análise desenvolvidas foram baseadas em cromatografia líquida de alta eficiência (HPLC) e técnicas de espectrometria de massas com ionização por Electrospray (ESI). A utilização da técnica de cromatografia líquida em escala preparativa permitiu o isolamento das amidas bis-fenilpropanoidicas, brasiliamida A e B e ainda, metabólitos secundários co-produzidos com estes compostos. A utilização do acoplamento entre as metodologias de análise LC/MS permitiu, através de estudos de fragmentação MS/MS, determinar condições otimizadas para a detecção seletiva das amidas nos meios de cultura, enriquecidos com diferentes fontes nitrogenadas. As análises quantitativas por HPLC/UV-MS/MS indicaram que o aminoácido L-fenilalanina intensifica a produção das brasiliamidas no extrato fúngico, e, a partir destes resultados, a rota biossintética dos fenilpropanóides produzidos pelo fungo P. brasilianum foi avaliada com a adição de [2-13C-Phe]. As análises dos extratos enriquecidos com aminoácido isotopicamente marcado via RMN 13C e HPLC/UV-MS, indicaram a incorporação de duas unidades do aminoácido L-fenilalanina na estrutura das brasiliamidas e a rota biossintética foi estabelecida a partir do caminho geral dos fenilpropanóides, metabólitos pouco reportados em fungos. A primeira etapa da rota biossintética foi avaliada usando extratos de ensaios enzimáticos com a enzima Fenilalanina Amônioliase (PAL), onde se visualizou a formação do ácido cinâmico a partir do aminoácido Phe através de monitoramento por HPLC/UV.Universidade Federal de Minas Geraisapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Química - PPGQUFSCarBRBiossínteseFungos endofíticosFenilpropanóideEspectrometria de massasFenilalanina amônia-liaseCIENCIAS EXATAS E DA TERRA::QUIMICAEstudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae)Biosynthetic study of phenylpropanoyl amides produced by The fungus Penicillium brasilianum, an endophyte from Melia azedarachinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-138736a86-d071-4ee1-8969-144f8d20b7ccinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL2236.pdfapplication/pdf4508455https://repositorio.ufscar.br/bitstream/ufscar/6444/1/2236.pdfda224f07c31e180bc11a5171a3f168f6MD51THUMBNAIL2236.pdf.jpg2236.pdf.jpgIM Thumbnailimage/jpeg8932https://repositorio.ufscar.br/bitstream/ufscar/6444/2/2236.pdf.jpg0fbccd5646550b544e307e8f53b8ad93MD52ufscar/64442023-09-18 18:31:11.511oai:repositorio.ufscar.br:ufscar/6444Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:11Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
dc.title.alternative.eng.fl_str_mv |
Biosynthetic study of phenylpropanoyl amides produced by The fungus Penicillium brasilianum, an endophyte from Melia azedarach |
title |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
spellingShingle |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) Fill, Taícia Pacheco Biossíntese Fungos endofíticos Fenilpropanóide Espectrometria de massas Fenilalanina amônia-liase CIENCIAS EXATAS E DA TERRA::QUIMICA |
title_short |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
title_full |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
title_fullStr |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
title_full_unstemmed |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
title_sort |
Estudos biossintéticos de amidas bis-fenilpropanoídicas produzidas pelo fungo Penicillium brasilianum, um endofítico de Melia azedarach (Meliaceae) |
author |
Fill, Taícia Pacheco |
author_facet |
Fill, Taícia Pacheco |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/0741638379707154 |
dc.contributor.author.fl_str_mv |
Fill, Taícia Pacheco |
dc.contributor.advisor1.fl_str_mv |
Rodrigues Filho, Edson |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3667941735597178 |
dc.contributor.authorID.fl_str_mv |
8e5be67e-894c-48a2-94a3-701e47160b77 |
contributor_str_mv |
Rodrigues Filho, Edson |
dc.subject.por.fl_str_mv |
Biossíntese Fungos endofíticos Fenilpropanóide Espectrometria de massas Fenilalanina amônia-liase |
topic |
Biossíntese Fungos endofíticos Fenilpropanóide Espectrometria de massas Fenilalanina amônia-liase CIENCIAS EXATAS E DA TERRA::QUIMICA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS EXATAS E DA TERRA::QUIMICA |
description |
In the present work it was developed analytical methodologies for investigation of biosynthesis of bis-phenylpropanoyl amides, using labeled precursors in the culture medium. The effect of different nitrogen sources on the bis-phenylpropanoyl amides production was also evaluated. These amides are reported as antiparasitary compounds and they also showed activity against gram-positive Bacillus subtilis in bioassays conducted at LaBioMMi (DQ/UFSCar). The analytical methodologies developed were based on High Performance Liquid Chromatography (HPLC) and Mass Spectrometry techniques, mainly electrospray ionization (ESI) and MS/MS. Using the methodologies developed for preparative liquid chromatography it were isolated the bis-phenylpropanoyl amides, brasiliamide A and B and other secondary metabolites co-produced. Liquid chromatography coupled with mass spectrometry techniques allowed fragmentation studies which lead to selective detection experiments of brasiliamides in mediums enhanced with different nitrogen sources. The quantitative analysis (HPLC/UV-MS/MS) indicated great influence of the amino acid L-phenylalanine on the bis-phenylpropanoyl amides production. From these results, the biosynthesis of the phenylapropanoids was investigated using [2-13CPhe]. The RMN 13C and HPLC/UV-MS analysis showed the incorporation of two amino acid units on brasiliamides structure formed by the phenylpropanoids pathway, which is uncommon in fungi. The first phenylpropanoid pathway step was evaluated throw using enzymatic extract with Phenylalanine ammoniolyase (PAL). The HPLC/UV analysis indicates the formation of cinnamic acid from Lphenylalanine. |
publishDate |
2009 |
dc.date.available.fl_str_mv |
2009-10-30 2016-06-02T20:36:21Z |
dc.date.issued.fl_str_mv |
2009-02-12 |
dc.date.accessioned.fl_str_mv |
2016-06-02T20:36:21Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FILL, Taícia Pacheco. Biosynthetic study of phenylpropanoyl amides produced by The fungus Penicillium brasilianum, an endophyte from Melia azedarach. 2009. 183 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2009. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/6444 |
identifier_str_mv |
FILL, Taícia Pacheco. Biosynthetic study of phenylpropanoyl amides produced by The fungus Penicillium brasilianum, an endophyte from Melia azedarach. 2009. 183 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2009. |
url |
https://repositorio.ufscar.br/handle/ufscar/6444 |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Universidade Federal de São Carlos |
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Programa de Pós-Graduação em Química - PPGQ |
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UFSCar |
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BR |
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Universidade Federal de São Carlos |
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