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Previous issue date: 2017-08-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O bioma Caatinga ? exclusivamente brasileiro, marcado pelo clima semi?rido. Este bioma se destaca pelo alto ?ndice de endemismo e desconhecimento cient?fico ao seu respeito. Para sobreviver neste ambiente, as plantas desenvolveram mecanismos intr?nsecos de percep??o dos sinais ambientais externos e suas respectivas respostas metab?licas. Dentre as esp?cies de ocorr?ncia no bioma Caatinga, como fonte de alcaloides bioativos, destaca-se Erythroxylum pungens, que apresenta potencial pouco explorado. Assim, o objetivo deste trabalho ? investigar o proteoma total e o perfil metab?lico de E. pungens, aliado ? bioprospec??o de alcaloides trop?nicos com potencial citot?xico. Para isso, no cap?tulo 1 identificou-se, por espectrometria de massas, sete alcaloides trop?nicos conhecidos; 3-(2-metilbutiriloxi)tropan-6, 7-diol como tamb?m 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 ra?zes de E. pungens. Al?m 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 c?lulas de tumorais de pr?stata. Para melhor conhecer a esp?cie e, com vistas na produ??o de alcaloides trop?nicos, trabalhou-se o proteoma e perfil alcalo?dico no cap?tulo 2. Dessa forma foi poss?vel identificar 1746 prote?nas nas folhas, 1779 no caule e 1026 na raiz de E. pungens. Atrav?s da recupera??o dos termos no Gene Ontology, observou-se os processos nos quais estas prote?nas est?o envolvidas, verificando o mesmo perfil de processos para os tr?s ?rg?os de E. pungens, com destaque para respostas a estresses bi?ticos, abi?ticos, qu?micos, al?m das respostas ao estresse oxidativo. Foram observados n?veis significativos de prote?nas envolvidas na fotoss?ntese das folhas de E. pungens, fotorrespira??o, al?m prote?nas relacionadas ?s respostas ao estresse: prote?nas da fam?lia 14-3-3, peroxidases, catalases, super?xido-dismutase e prote?nas de choque t?rmico, que tamb?m podem atuar no turnover prote?co. Ainda foram identificadas cinco prote?nas hom?logas a tropinona redutase na folha, e produ??o de alcaloide em todos os ?rg?os. No cap?tulo 3, por meio do experimento com esp?cimes de E. pungens submetidas a estresse h?drico por suspens?o de rega em casa de vegeta??o, notou-se que esta esp?cie investe no aumento de prote?nas relacionadas ?s respostas ao estresse h?drico, aumenta significativamente o teor de prolina livre no citosol. Ademais, mat?m a produ??o de alcaloides que parecem estar mais relacionado a fase de desenvolvimento da planta. Com isso, os resultados deste trabalho busca contribuir para a fitoqu?mica e fisiologia de E. pungens, assim como promove o uso racional da esp?cie, e consequentemente a conserva??o do bioma Caatinga. / 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.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/24802 |
| Date | 25 August 2017 |
| Creators | Pereira, Gabrielle Macedo |
| Contributors | 00104164042, Uchoa, Adriana Ferreira, 58024867320, Voigt, Eduardo Luiz, 93217617991, Fett Neto, Arthur Germano, 48891940020, Lago, Jo?o Henrique Ghilardi, 26413371854, Pedrosa, Matheus de Freitas Fernandes, Giordani, Raquel Brandt |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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