Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/6331 |
Resumo: | The PhD work aimed the exploration of fungal metabolism of NICL3 and NICL5, both initially identified as Xylaria sp., isolated as endophytic fungi from Cupressus lusitanica. The investigation of the NICL3 metabolism allowed the isolation and identification of new compounds, one unpublished eremophilane sesquiterpene and a mixture of two cyclic peptides. In addition, from NICL5 were isolated cytochalasins C, D, Q, 18- desoxy cytochalasin Q and trichodepsipeptide. The production of this depsipeptide by NICL5 is very relevant, since there is only one paper in the literature that reports the production of this class of compounf by fungi belonging to Xylaria genus. Another aspect applied in this work was the use of OSMAC approach, through supplementation of different nitrogen sources in the culture medium of NICL5, to obtain new cytochalasins. However, only the administration of NH4Br was promising since it allowed the isolation of 4- hydroxymethyl cytochalasin C a new metabolite. The secondary metabolism NICL5 is strongly guided to the production of cytochalasins C, D and Q, independently of the nitrogen source administered in the culture medium, suggesting that the microorganism is able to perform de novo biosynthesis. Mass spectrometry studies demonstrated that administration of Na15NO3, 15NH4Cl and 15N-glycine led to incorporation of labeled nitrogen into cytochalasin D. Supplementation of the culture medium with 2-13C glycine showed the incorporation of labeled carbon in positions in which the enzyme S-adenosyl methionine is responsible for introducing two methyl groups and one methylene. However, genomic and proteomic studies are essential to prove this process. The NICL5 fungus has an interesting enzymatic potential because it was able to promote reductions of carboxylic acid into alcohol and performs the incorporation of GABA and glucose to the substrate core, when the substrate was p-bromo benzoic acid. This enzymatic potencial appears to be closely related to the eletronic nature of the substrate, since similar reaction occurred with p-cloro benzoic acid and bioreduction occurred also with p-nitro benzoic acid. A collection of fungi belonging to Xylariaceae family, which were identified according to molecular techniques was created. Interestingly NICL3 was classified as Nemania aff. primolutea and NICL5 as Xylaria aff. arbuscula. The identification techniques based on molecular biology, biomarkers profile by MALDI-TOF and metabolic profile by high resolution mass spectrometry of all members of the collection were confronted, and it was verified a good level of agreement between three different techniques. |
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Amaral, Luciana da SilvaRodrigues Filho, Edsonhttp://lattes.cnpq.br/3667941735597178http://lattes.cnpq.br/222094260724935102a1660a-0a63-4b02-913b-3ca9ca3ad1e22016-06-02T20:34:59Z2015-03-022016-06-02T20:34:59Z2014-11-11AMARAL, Luciana da Silva. Metabolomic and biosynthetic studies for the chemical and taxonomic contribution of xylariaceae family microorganisms. 2014. 1 f. Tese (Doutorado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2014.https://repositorio.ufscar.br/handle/ufscar/6331The PhD work aimed the exploration of fungal metabolism of NICL3 and NICL5, both initially identified as Xylaria sp., isolated as endophytic fungi from Cupressus lusitanica. The investigation of the NICL3 metabolism allowed the isolation and identification of new compounds, one unpublished eremophilane sesquiterpene and a mixture of two cyclic peptides. In addition, from NICL5 were isolated cytochalasins C, D, Q, 18- desoxy cytochalasin Q and trichodepsipeptide. The production of this depsipeptide by NICL5 is very relevant, since there is only one paper in the literature that reports the production of this class of compounf by fungi belonging to Xylaria genus. Another aspect applied in this work was the use of OSMAC approach, through supplementation of different nitrogen sources in the culture medium of NICL5, to obtain new cytochalasins. However, only the administration of NH4Br was promising since it allowed the isolation of 4- hydroxymethyl cytochalasin C a new metabolite. The secondary metabolism NICL5 is strongly guided to the production of cytochalasins C, D and Q, independently of the nitrogen source administered in the culture medium, suggesting that the microorganism is able to perform de novo biosynthesis. Mass spectrometry studies demonstrated that administration of Na15NO3, 15NH4Cl and 15N-glycine led to incorporation of labeled nitrogen into cytochalasin D. Supplementation of the culture medium with 2-13C glycine showed the incorporation of labeled carbon in positions in which the enzyme S-adenosyl methionine is responsible for introducing two methyl groups and one methylene. However, genomic and proteomic studies are essential to prove this process. The NICL5 fungus has an interesting enzymatic potential because it was able to promote reductions of carboxylic acid into alcohol and performs the incorporation of GABA and glucose to the substrate core, when the substrate was p-bromo benzoic acid. This enzymatic potencial appears to be closely related to the eletronic nature of the substrate, since similar reaction occurred with p-cloro benzoic acid and bioreduction occurred also with p-nitro benzoic acid. A collection of fungi belonging to Xylariaceae family, which were identified according to molecular techniques was created. Interestingly NICL3 was classified as Nemania aff. primolutea and NICL5 as Xylaria aff. arbuscula. The identification techniques based on molecular biology, biomarkers profile by MALDI-TOF and metabolic profile by high resolution mass spectrometry of all members of the collection were confronted, and it was verified a good level of agreement between three different techniques.O trabalho de doutorado visou a exploração do metabolismo fúngico de NICL3 e NICL5, ambos inicialmente identificados como Xylaria sp., isolados como endofítico de Cupressus lusitanica. A investigação do metabolismo dos fungos permitiu o isolamento e identificação de novos metabólitos, pois NICL3 produziu um novo sesquiterpeno eremofilano e uma mistura de dois peptídeos cíclicos. Já de NICL5 foram isolados além das citocalasinas C, D e Q, a 18-desoxicitolasina Q e o tricodepsipeptídeo. O isolamento deste depsipetídeo por NICL5 é algo muito relevante, dado que na literatura há somente um trabalho que relata a produção desta classe de composto por fungos do gênero Xylaria. Outra vertente aplicada neste trabalho foi o emprego da abordagem OSMAC, através da suplementação de diferentes fontes nitrogenadas ao meio de cultivo de NICL5, visando obter novas citocalasinas. No entanto, somente a administração de NH4Br se mostrou promissora, pois permitiu o isolamento da 4-hidroximetil citocalasina C, metabólito inédito. O metabolismo secundário de NICL5 se mostrou fortemente direcionado para a produção das citocalasinas C, D e Q, independente da fonte nitrogenada administrada ao meio de cultura, o que sugeriu que o microorganismo fosse capaz de realizar biossíntese de novo destes metabólitos. Estudos por espectrometria de massas mostraram que a administração de Na15NO3, 15NH4Cl e 15N-glicina levou a incorporação do nitrogênio marcado na citocalasina D. Já a suplementação do meio de cultivo do micro-organismo com 2-13C glicina apontou a incorporação do carbono marcado nas posições em que a enzima S-adenosil metionina é responsável por introduzir dois grupos metilas e um metileno. Entretanto, estudos genômico e proteômico são fundamentais para comprovar este processo. O fungo NICL5 possui um potencial enzimático muito interessante, pois promoveu a redução do ácido carboxílico ao álcool correspondente, além de realizar a incorporação do GABA e glicose ao substrato. Este potencial enzimático parece estar intimamente relacionado com a natureza eletrônica do substrato, pois reações semelhantes aconteceram com o ácido p-cloro benzoico e a redução do ácido p-nitro benzoico ao álcool. Foi criada uma coleção de fungos da família Xylariaceae, os quais foram identificados de acordo com técnicas moleculares. Curiosamente NICL3 foi classificado como Nemania aff. primolutea e NICL5 como Xylaria aff. arbuscula. Foram confrontadas as técnicas de identificação baseadas em biologia molecular, perfil de biomoléculas por MALDI-TOF e o perfil metabólico por espectrometria de massas de alta resolução de todos os representantes da coleção de Xylariaceae, pôde-se verificar um bom nível de concordância entre as diferentes técnicas.Universidade Federal de Sao Carlosapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Química - PPGQUFSCarBRQuímicaFungo endofíticoCitocalasinasBiossínteseQuímica microbianaCIENCIAS EXATAS E DA TERRA::QUIMICAEstudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família XylariaceaeMetabolomic and biosynthetic studies for the chemical and taxonomic contribution of xylariaceae family microorganismsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-1-138736a86-d071-4ee1-8969-144f8d20b7ccinfo:eu-repo/semantics/embargoedAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL6518.pdf6518.pdfapplication/pdf16307915https://repositorio.ufscar.br/bitstream/ufscar/6331/1/6518.pdf98e4b84f5bb3b56375bc9ef8c6490150MD51TEXT6518.pdf.txt6518.pdf.txtExtracted texttext/plain0https://repositorio.ufscar.br/bitstream/ufscar/6331/2/6518.pdf.txtd41d8cd98f00b204e9800998ecf8427eMD52THUMBNAIL6518.pdf.jpg6518.pdf.jpgIM Thumbnailimage/jpeg9865https://repositorio.ufscar.br/bitstream/ufscar/6331/3/6518.pdf.jpga1fc69070e49143b9d20844fc7e99a11MD53ufscar/63312023-09-18 18:31:15.408oai:repositorio.ufscar.br:ufscar/6331Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:15Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
dc.title.alternative.eng.fl_str_mv |
Metabolomic and biosynthetic studies for the chemical and taxonomic contribution of xylariaceae family microorganisms |
title |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
spellingShingle |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae Amaral, Luciana da Silva Química Fungo endofítico Citocalasinas Biossíntese Química microbiana CIENCIAS EXATAS E DA TERRA::QUIMICA |
title_short |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
title_full |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
title_fullStr |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
title_full_unstemmed |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
title_sort |
Estudos metabolômicos e biossintéticos visando contribuições para a química e taxonomia de microorganismos da família Xylariaceae |
author |
Amaral, Luciana da Silva |
author_facet |
Amaral, Luciana da Silva |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/2220942607249351 |
dc.contributor.author.fl_str_mv |
Amaral, Luciana da Silva |
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 |
02a1660a-0a63-4b02-913b-3ca9ca3ad1e2 |
contributor_str_mv |
Rodrigues Filho, Edson |
dc.subject.por.fl_str_mv |
Química Fungo endofítico Citocalasinas Biossíntese Química microbiana |
topic |
Química Fungo endofítico Citocalasinas Biossíntese Química microbiana CIENCIAS EXATAS E DA TERRA::QUIMICA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS EXATAS E DA TERRA::QUIMICA |
description |
The PhD work aimed the exploration of fungal metabolism of NICL3 and NICL5, both initially identified as Xylaria sp., isolated as endophytic fungi from Cupressus lusitanica. The investigation of the NICL3 metabolism allowed the isolation and identification of new compounds, one unpublished eremophilane sesquiterpene and a mixture of two cyclic peptides. In addition, from NICL5 were isolated cytochalasins C, D, Q, 18- desoxy cytochalasin Q and trichodepsipeptide. The production of this depsipeptide by NICL5 is very relevant, since there is only one paper in the literature that reports the production of this class of compounf by fungi belonging to Xylaria genus. Another aspect applied in this work was the use of OSMAC approach, through supplementation of different nitrogen sources in the culture medium of NICL5, to obtain new cytochalasins. However, only the administration of NH4Br was promising since it allowed the isolation of 4- hydroxymethyl cytochalasin C a new metabolite. The secondary metabolism NICL5 is strongly guided to the production of cytochalasins C, D and Q, independently of the nitrogen source administered in the culture medium, suggesting that the microorganism is able to perform de novo biosynthesis. Mass spectrometry studies demonstrated that administration of Na15NO3, 15NH4Cl and 15N-glycine led to incorporation of labeled nitrogen into cytochalasin D. Supplementation of the culture medium with 2-13C glycine showed the incorporation of labeled carbon in positions in which the enzyme S-adenosyl methionine is responsible for introducing two methyl groups and one methylene. However, genomic and proteomic studies are essential to prove this process. The NICL5 fungus has an interesting enzymatic potential because it was able to promote reductions of carboxylic acid into alcohol and performs the incorporation of GABA and glucose to the substrate core, when the substrate was p-bromo benzoic acid. This enzymatic potencial appears to be closely related to the eletronic nature of the substrate, since similar reaction occurred with p-cloro benzoic acid and bioreduction occurred also with p-nitro benzoic acid. A collection of fungi belonging to Xylariaceae family, which were identified according to molecular techniques was created. Interestingly NICL3 was classified as Nemania aff. primolutea and NICL5 as Xylaria aff. arbuscula. The identification techniques based on molecular biology, biomarkers profile by MALDI-TOF and metabolic profile by high resolution mass spectrometry of all members of the collection were confronted, and it was verified a good level of agreement between three different techniques. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-11-11 |
dc.date.available.fl_str_mv |
2015-03-02 2016-06-02T20:34:59Z |
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2016-06-02T20:34:59Z |
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info:eu-repo/semantics/publishedVersion |
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doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
AMARAL, Luciana da Silva. Metabolomic and biosynthetic studies for the chemical and taxonomic contribution of xylariaceae family microorganisms. 2014. 1 f. Tese (Doutorado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2014. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/6331 |
identifier_str_mv |
AMARAL, Luciana da Silva. Metabolomic and biosynthetic studies for the chemical and taxonomic contribution of xylariaceae family microorganisms. 2014. 1 f. Tese (Doutorado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2014. |
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https://repositorio.ufscar.br/handle/ufscar/6331 |
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Universidade Federal de São Carlos |
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BR |
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