CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Amburana cearensis (cumaru) Ã uma leguminosa subutilizada da Caatinga. O alto teor proteico das sementes faz delas uma possÃvel nova fonte de proteÃnas alimentares. Foram determinadas as melhores condiÃÃes de extraÃÃo proteica para posterior produÃÃo do isolado proteico de cumaru (IPAc). As anÃlises tambÃm foram conduzidas com a farinha delipidada (FDAc) e com um isolado proteico de soja comercial (IPS). IPAc apresentou teor proteico > 90% e composiÃÃo de aminoÃcidos comparÃvel a IPS, com notÃvel deficiÃncia em metionina. Foram determinadas algumas propriedades funcionais para predizer sua adequaÃÃo como ingrediente na indÃstria alimentÃcia. Os resultados para FDAc, IPAc e IPS foram, respectivamente: solubilidade 20-79; 9-99 e 3-76%; capacidade de retenÃÃo de Ãgua 1,7; 3,9 e 7,5 g/g; capacidade de retenÃÃo de Ãleo 1,5; 6,3 e 1,6 g/g; capacidade emulsificante / estabilidade 52 / 46%; 55 / 51% e 53 / 21%; capacidade espumante / estabilidade 47 / 32%; 65 / 53% e 33 / 8 e menor concentraÃÃo geleificante 18, 16 e 12%. Para investigar se IPAc e IPS apresentam indÃcios de toxicidade, cÃlulas de adenocarcinoma humano foram expostas por 24 h aos isolados. Para comparaÃÃo, tambÃm foi realizada exposiÃÃo com os isolados de soja (IPGm), feijÃo comum (IPPv), feijÃo de porco (IPCe) e mamona (IPRc), a fraÃÃo proteica de mamona (FPRc), as lectinas PHA-E e ConA e o controle quÃmico (Crtl_Quim). O RNA das cÃlulas foi hibridizado em microarranjos contendo o genoma humano completo. Os perfis de expressÃo gÃnica apÃs exposiÃÃo Ãs proteÃnas foram comparados aos de PBS e Crtl_Quim e agrupados hierarquicamente. Crtl_Quim, IPAc e IPGm se mostraram inadequados para a continuaÃÃo das anÃlises por provocar um efeito superestimado. A anÃlise de vias e processos biolÃgicos afetados apontou que genes envolvidos com a biossÃntese de colesterol, estresse do RE e resposta imune foram superexpressos e genes envolvidos com o ciclo celular e replicaÃÃo do DNA foram subexpressos apÃs exposiÃÃo a IPPv, IPCe, PHA-E e ConA. As amostras IPRc e FPRc regularam para cima genes envolvidos com a apoptose e resposta imune e para baixo genes envolvidos com o ciclo celular e sinalizaÃÃo do cAMP. AnÃlise do mapa conectivo mostrou alta correlaÃÃo de ConA, PHA-E e IPCe com drogas bloqueadoras de canais de Ca2+ e de K+; enquanto IPRc e FPRc apresentam funÃÃo biolÃgica semelhante a pelo menos seis drogas inibitÃrias da sÃntese proteica. Conclui-se que IPAc tem grande potencialidade como novo alimento proteico com grande aplicabilidade na indÃstria de alimentos, devendo, no entanto, sofrer modificaÃÃes no mÃtodo de obtenÃÃo visando a melhora na solubilidade e a eliminaÃÃo do sal residual. / Amburana cearensis (cumaru) is an underutilized legume from Caatinga. Because of their high protein content its seeds are a possible novel source of food protein. Were determined the best conditions for protein extraction and preparation of protein isolate (IPAc).Analysis were carried out also with the defatted flour (FDAc) and with a marketed soybean protein isolate (IPS). IPAc showed protein content of 90% and amino acid composition compatible with the IPSâs composition, with remarkable low methionine content. Some functional properties were measured in order to predict the adequacy of the isolates as an ingredient for food industry. The results for FDAc, IPAc and IPS were, respectively: solubility 20-79; 9-99 e 3-76%; water holding capacity 1.7; 3.9 e 7.5 g/g; oil holding capacity 1.5; 6.3 e 1.6 g/g; emulsion formation / stability 52 / 46%; 55 / 51% e 53 / 21%; foamability / stability 47 / 32%; 65 / 53% e 33 / 8; least gelling concentration (LGC) 18, 16 and 12%. In order to investigate whether IPAc e IPS exhibit signs of toxicity, two cell lines of human adenocarcinoma (MCF-7 e Caco-2) were exposed to these isolates as well as to others prepared from soyabean (IPGm), white bean (IPPv), Jack bean (IPCe) and castor bean (IPRc), to the protein fraction from castor bean (FPRc), to the lectins PHA-E and ConA (50 Âg/mL) and to the chemical control (Crtl_Quim). Subcitotoxic doses were determined by viability test to later exposure assay for 24 h. The MCF-7 cellâs RNA were extracted and hybridized to a microarray containing the complete human genome. The expression profiles after the exposition to the proteins were compared to that ones generated after PBS and Crtl_Quim and hierarchicaly clustered. Crtl_Quim, IPAc and IPGm were unsuitable for further analysis by their overestimated effect. Analysis of biological pathways and process showed that genes involved with the cholesterol biosynthesis, ER stress and immune response were upregulated and genes involved with cell cycle and DNA replication were downregulated after exposure of cells to IPPv, IPCe, PHA-E and ConA. The samples IPRc and FPRc upregulated genes involved with apoptosis and immune response and downregulated genes involved with cell cycle and cAMP signaling. Connectivity Map analysis showed high correlation of ConA, PHA-E and IPCe with drugs that are Ca2+ and K+ channel blockers, while IPRc and FPRc showed biological function similar to at least six drugs inhibitors of protein synthesis. Itâs possible to conclude that the IPAc has a high potentiality as a novel protein food with wide applicability in the food industry and should, however, be modified in the method of obtaining aiming the improvement in its solubility and eliminating the residual salt.
| Identifer | oai:union.ndltd.org:IBICT/oai:www.teses.ufc.br:8945 |
| Date | 09 May 2014 |
| Creators | Nathanna Mateus de Sousa |
| Contributors | Ana de FÃtima Fontenele Urano Carvalho, Adrianus Antonius Cornelis Maria Peijnenburg, Helena Alves de Carvalho Sampaio, Ilka Maria Vasconcelos, Renato de Azevedo Moreira |
| Publisher | Universidade Federal do CearÃ, Programa de PÃs-GraduaÃÃo em BioquÃmica, UFC, BR |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFC, instname:Universidade Federal do Ceará, instacron:UFC |
| Rights | info:eu-repo/semantics/openAccess |
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