Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos

Caatinga biome is exclusive brazilian, marked by semi-arid climate. This biome stands out for the high endemism index and scientific unfamiliarity.To survive in this environment with peculiar edaphoclimatic conditions, plants show evolution of intrinsic mechanisms of external environmental signals p...

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Autor principal: Pereira, Gabrielle Macedo
Outros Autores: Giordani, Raquel Brandt
Formato: doctoralThesis
Idioma:por
Publicado em: Brasil
Assuntos:
Erythroxylaceae
Metabólitos secundários
Caatinga
Proteômica
Estresses ambientais
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
Endereço do item:https://repositorio.ufrn.br/jspui/handle/123456789/24802
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id ri-123456789-24802
record_format dspace
institution Repositório Institucional
collection RI - UFRN
language por
topic Erythroxylaceae
Metabólitos secundários
Caatinga
Proteômica
Estresses ambientais
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
spellingShingle Erythroxylaceae
Metabólitos secundários
Caatinga
Proteômica
Estresses ambientais
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
Pereira, Gabrielle Macedo
Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
description Caatinga biome is exclusive brazilian, marked by semi-arid climate. This biome stands out for the high endemism index and scientific unfamiliarity.To survive in this environment with peculiar edaphoclimatic conditions, plants show evolution of intrinsic mechanisms of external environmental signals perception and their respectives metabolic responses. Among the occurrence genera in Caatinga biome, as bioactive alkaloids source, the Erythroxylum pungens species, which has unexplored potential, is outstanding. Thus, the objective of this work is to investigate total proteome and metabolic fingerprint of E. pungens, together with tropane alkaloids bioprospection with cytotoxic potential. For this, in chapter 1 it was possible to identify, by mass spectrometry, seven known tropane alkaloids; 3-(2-methylbutyryloxy)tropan-6, 7-diol as well as 3-(2-methylbutyryloxy)tropan-6, 7-diol were isolated and characterized by 1D and 2D NMR for the first time. N,N-Dimethyl-1-H-indol-3-ethanamine was isolated and characterized from roots of E. punctures. In addition, it was found that alkaloids isolated from E. pungens, against five cell lines, indicated selective potential of 3-(2-methylbutyryloxy)tropan-6, 7-diol against prostate tumor cells. In chapter 2, it was possible to identify 1746 proteins in the leaves, 1779 in the stem and 1026 in the root of E. pungens. Through the recovery of the GO terms, it was possible to evaluate processes in which these proteins are involved, verifying the same process profile for three organs of E. pungens, with emphasis on biotic, abiotic, chemical stress responses, as well as responses to oxidative stress. Were observed significant proteins levels involved in photosynthesis of E. pungens leaves, photorespiration, as well as many proteins related to stress responses: proteins from 14-3-3 family, peroxidases, catalases, superoxide dismutase and shock proteins, which can also act on protein turnover. Were still identificated five proteins homologous to tropinone reductase, and alkaloid production in all organs suggesting that even under chronic stress conditions, E. pungens maintains the production of these metabolites. In chapter 3, through specimens submission experiment of E. pungens to water stress by irrigation suspension in greenhouse, it was observed that this species invest in the increase of proteins related to the responses to water stress, it significantly increases free proline content in cytosol which functions as an important osmoprotector. Furthermore, maintains alkaloids production that appear to be more related to development stage of the plant than stress condition in which it is inserted. The results of this work seek to contribute to knowledge of E. pungens phytochemistry and physiology, as well as to promote the species and consequently conservation of Caatinga biome.
author2 Giordani, Raquel Brandt
author_facet Giordani, Raquel Brandt
Pereira, Gabrielle Macedo
format doctoralThesis
author Pereira, Gabrielle Macedo
author_sort Pereira, Gabrielle Macedo
title Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
title_short Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
title_full Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
title_fullStr Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
title_full_unstemmed Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos
title_sort erythroxylum pungens o. e. shulz: proteômica total e bioprospecção de alcaloides tropânicos
publisher Brasil
publishDate 2018
url https://repositorio.ufrn.br/jspui/handle/123456789/24802
work_keys_str_mv AT pereiragabriellemacedo erythroxylumpungensoeshulzproteomicatotalebioprospeccaodealcaloidestropanicos
AT pereiragabriellemacedo erythroxylumpungensoeshulztotalproteomicsandbioprospectionoftropanealkaloids
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spelling ri-123456789-248022022-11-04T19:42:01Z Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos Erythroxylum pungens O. E. shulz: total proteomics and bioprospection of tropane alkaloids Pereira, Gabrielle Macedo Giordani, Raquel Brandt http://lattes.cnpq.br/7975202296724439 http://lattes.cnpq.br/5032750980466610 Pedrosa, Matheus de Freitas Fernandes http://lattes.cnpq.br/2929963416385218 Uchoa, Adriana Ferreira https://orcid.org/0000-0002-3822-6502 http://lattes.cnpq.br/6644671747055211 Voigt, Eduardo Luiz http://lattes.cnpq.br/2455621187263790 Fett Neto, Arthur Germano https://orcid.org/0000-0001-5477-2610 http://lattes.cnpq.br/6760789832926234 Lago, João Henrique Ghilardi https://orcid.org/0000-0002-1193-8374 http://lattes.cnpq.br/2325513222088331 Erythroxylaceae Metabólitos secundários Caatinga Proteômica Estresses ambientais CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA Caatinga biome is exclusive brazilian, marked by semi-arid climate. This biome stands out for the high endemism index and scientific unfamiliarity.To survive in this environment with peculiar edaphoclimatic conditions, plants show evolution of intrinsic mechanisms of external environmental signals perception and their respectives metabolic responses. Among the occurrence genera in Caatinga biome, as bioactive alkaloids source, the Erythroxylum pungens species, which has unexplored potential, is outstanding. Thus, the objective of this work is to investigate total proteome and metabolic fingerprint of E. pungens, together with tropane alkaloids bioprospection with cytotoxic potential. For this, in chapter 1 it was possible to identify, by mass spectrometry, seven known tropane alkaloids; 3-(2-methylbutyryloxy)tropan-6, 7-diol as well as 3-(2-methylbutyryloxy)tropan-6, 7-diol were isolated and characterized by 1D and 2D NMR for the first time. N,N-Dimethyl-1-H-indol-3-ethanamine was isolated and characterized from roots of E. punctures. In addition, it was found that alkaloids isolated from E. pungens, against five cell lines, indicated selective potential of 3-(2-methylbutyryloxy)tropan-6, 7-diol against prostate tumor cells. In chapter 2, it was possible to identify 1746 proteins in the leaves, 1779 in the stem and 1026 in the root of E. pungens. Through the recovery of the GO terms, it was possible to evaluate processes in which these proteins are involved, verifying the same process profile for three organs of E. pungens, with emphasis on biotic, abiotic, chemical stress responses, as well as responses to oxidative stress. Were observed significant proteins levels involved in photosynthesis of E. pungens leaves, photorespiration, as well as many proteins related to stress responses: proteins from 14-3-3 family, peroxidases, catalases, superoxide dismutase and shock proteins, which can also act on protein turnover. Were still identificated five proteins homologous to tropinone reductase, and alkaloid production in all organs suggesting that even under chronic stress conditions, E. pungens maintains the production of these metabolites. In chapter 3, through specimens submission experiment of E. pungens to water stress by irrigation suspension in greenhouse, it was observed that this species invest in the increase of proteins related to the responses to water stress, it significantly increases free proline content in cytosol which functions as an important osmoprotector. Furthermore, maintains alkaloids production that appear to be more related to development stage of the plant than stress condition in which it is inserted. The results of this work seek to contribute to knowledge of E. pungens phytochemistry and physiology, as well as to promote the species and consequently conservation of Caatinga biome. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) O bioma Caatinga é exclusivamente brasileiro, marcado pelo clima semiárido. Este bioma se destaca pelo alto índice de endemismo e desconhecimento científico ao seu respeito. Para sobreviver neste ambiente, as plantas desenvolveram mecanismos intrínsecos de percepção dos sinais ambientais externos e suas respectivas respostas metabólicas. Dentre as espécies de ocorrência no bioma Caatinga, como fonte de alcaloides bioativos, destaca-se Erythroxylum pungens, que apresenta potencial pouco explorado. Assim, o objetivo deste trabalho é investigar o proteoma total e o perfil metabólico de E. pungens, aliado à bioprospecção de alcaloides tropânicos com potencial citotóxico. Para isso, no capítulo 1 identificou-se, por espectrometria de massas, sete alcaloides tropânicos conhecidos; 3-(2-metilbutiriloxi)tropan-6, 7-diol como também 3-(2-metilbutiriloxi)nortropan-6, 7-diol foram isolados e caracterizado por RMN 1D e 2D pela primeira vez. N,N-Dimetil-1-H-indol-3-etanamina foi isolado e caracterizado a partir de raízes de E. pungens. Além disso, verificou-se que alcaloides isolados de E. pungens, frente a cinco linhagens celulares, apontaram potencial seletivo de 3-(2-metilbutiriloxi)tropan-6, 7-diol contra células de tumorais de próstata. Para melhor conhecer a espécie e, com vistas na produção de alcaloides tropânicos, trabalhou-se o proteoma e perfil alcaloídico no capítulo 2. Dessa forma foi possível identificar 1746 proteínas nas folhas, 1779 no caule e 1026 na raiz de E. pungens. Através da recuperação dos termos no Gene Ontology, observou-se os processos nos quais estas proteínas estão envolvidas, verificando o mesmo perfil de processos para os três órgãos de E. pungens, com destaque para respostas a estresses bióticos, abióticos, químicos, além das respostas ao estresse oxidativo. Foram observados níveis significativos de proteínas envolvidas na fotossíntese das folhas de E. pungens, fotorrespiração, além proteínas relacionadas às respostas ao estresse: proteínas da família 14-3-3, peroxidases, catalases, superóxido-dismutase e proteínas de choque térmico, que também podem atuar no turnover proteíco. Ainda foram identificadas cinco proteínas homólogas a tropinona redutase na folha, e produção de alcaloide em todos os órgãos. No capítulo 3, por meio do experimento com espécimes de E. pungens submetidas a estresse hídrico por suspensão de rega em casa de vegetação, notou-se que esta espécie investe no aumento de proteínas relacionadas às respostas ao estresse hídrico, aumenta significativamente o teor de prolina livre no citosol. Ademais, matém a produção de alcaloides que parecem estar mais relacionado a fase de desenvolvimento da planta. Com isso, os resultados deste trabalho busca contribuir para a fitoquímica e fisiologia de E. pungens, assim como promove o uso racional da espécie, e consequentemente a conservação do bioma Caatinga. 2018-02-23T21:04:51Z 2018-02-23T21:04:51Z 2017-08-25 doctoralThesis PEREIRA, Gabrielle Macedo. Erythroxylum pungens O. E. shulz: proteômica total e bioprospecção de alcaloides tropânicos. 2017. 156f. Tese (Doutorado em Bioquímica) - Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, 2017. https://repositorio.ufrn.br/jspui/handle/123456789/24802 por Acesso Aberto application/pdf Brasil UFRN PROGRAMA DE PÓS-GRADUAÇÃO EM BIOQUÍMICA

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