Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/20675 |
Resumo: | The yam is an important tuber in Brazil Northeast food culture, also being considered a functional food, as it has proteins with potential bioactive properties. The objective of this study was to characterize the peptide profile and perform the bioprospection of bioactive peptides obtained from in vitro simulated gastrointestinal digestion of yam proteins (Dioscorea cayennensis) grown in Brazil. Therefore, the sample obtained was initially analyzed for partial chemical composition, followed by obtaining the flour for the yam extract and protein concentrate (YPC) preparation, submitted to simulated digestion in vitro, and finally the characterization and profile of the obtained peptides were performed. The protein extract was subjected to isoelectric precipitation to obtain the YPC, which was then dialyzed and freeze-dried. The YPC was submitted to simulated gastrointestinal (GI) digestion, generating two phases: the gastric phase hydrolysate (GPH) and continuous hydrolysis until the gastrointestinal phase hydrolysate (GIPH). The samples: YPC, GPH and GIPH were submitted to analysis of protein and peptide profile: total amino acids and free amino acids (FAA); molecular mass (MM); hydrophobicity by reverse phase chromatography and electrophoresis (Tricine-SDS-PAGE); protein profile (two-dimensional gel electrophoresis) and peptide profile (nano-LC-ESI-MS/MS, MALDI-TOF). The identified peptides were submitted to the bioactivity prediction through the Bioactive Peptides Database (BIOPEP) and PeptideRanker (inhibitory activity of the angiotensin-converting enzyme-ACE). To validate results obtained in silico, analysis were performed to determine biological activities in vitro: antioxidant (DPPH, ABTS, ORAC and protection from DNA damage), antibacterial and ACE’s inhibitor. The results of 2D-electrophoresis show the presence of the dioscorin protein, as the yam’s major protein. The YCP resulted in 64.0 % protein, with all essential amino acids with an appropriate score according to FAO, except sulfur amino acids. After the GI digestion of the YPC, a significant increase in the release (p <0.5) of AAL was evidenced. The efficiency of GI digestion was verified according to the distribution profile of MW, with approximately 71.0 % of peptides with MW <2 kDa generated in the GIPH. Most of the identified peptides belong to the D. cayennensis dioscorin protein. GPH peptides had their MM decreased with the progression of GI hydrolysis, resulting in smaller MM peptides in GIPH. In addition, sequences with potential for bioactive peptides, with antihypertensive, antidiabetic, antioxidant and antibacterial activity were identified from the BIOPEP prediction. The presence of a peptide sequence containing proline and leucine (LAPLPL) was found in the GPH and GIPH, which can contribute to the inhibition of ACE, as well as peptides containing aromatic residues in the C-terminal (DITWT and FLSWT) that allow the donation of electrons and stabilization of free radicals. These findings suggest that these characteristics contributed to the antioxidant activity in all in vitro assays. Hydrophobic and positively charged peptides (IFDQTLGKLR) may have made it possible to inhibit the growth of the bacteria Escherichia coli. Therefore, the peptides obtained after the GI digestion of yam proteins can contribute to the prevention of infections and chronic degenerative diseases development, with an impact on human health. |
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Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitroProteína vegetalPeptídeos antibacterianosPeptídeos antioxidantesPeptídeos anti-hipertensivosBIOPEPTuber proteinAntibacterial peptidesAntioxidant peptidesAntihypertensive peptidesCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOSThe yam is an important tuber in Brazil Northeast food culture, also being considered a functional food, as it has proteins with potential bioactive properties. The objective of this study was to characterize the peptide profile and perform the bioprospection of bioactive peptides obtained from in vitro simulated gastrointestinal digestion of yam proteins (Dioscorea cayennensis) grown in Brazil. Therefore, the sample obtained was initially analyzed for partial chemical composition, followed by obtaining the flour for the yam extract and protein concentrate (YPC) preparation, submitted to simulated digestion in vitro, and finally the characterization and profile of the obtained peptides were performed. The protein extract was subjected to isoelectric precipitation to obtain the YPC, which was then dialyzed and freeze-dried. The YPC was submitted to simulated gastrointestinal (GI) digestion, generating two phases: the gastric phase hydrolysate (GPH) and continuous hydrolysis until the gastrointestinal phase hydrolysate (GIPH). The samples: YPC, GPH and GIPH were submitted to analysis of protein and peptide profile: total amino acids and free amino acids (FAA); molecular mass (MM); hydrophobicity by reverse phase chromatography and electrophoresis (Tricine-SDS-PAGE); protein profile (two-dimensional gel electrophoresis) and peptide profile (nano-LC-ESI-MS/MS, MALDI-TOF). The identified peptides were submitted to the bioactivity prediction through the Bioactive Peptides Database (BIOPEP) and PeptideRanker (inhibitory activity of the angiotensin-converting enzyme-ACE). To validate results obtained in silico, analysis were performed to determine biological activities in vitro: antioxidant (DPPH, ABTS, ORAC and protection from DNA damage), antibacterial and ACE’s inhibitor. The results of 2D-electrophoresis show the presence of the dioscorin protein, as the yam’s major protein. The YCP resulted in 64.0 % protein, with all essential amino acids with an appropriate score according to FAO, except sulfur amino acids. After the GI digestion of the YPC, a significant increase in the release (p <0.5) of AAL was evidenced. The efficiency of GI digestion was verified according to the distribution profile of MW, with approximately 71.0 % of peptides with MW <2 kDa generated in the GIPH. Most of the identified peptides belong to the D. cayennensis dioscorin protein. GPH peptides had their MM decreased with the progression of GI hydrolysis, resulting in smaller MM peptides in GIPH. In addition, sequences with potential for bioactive peptides, with antihypertensive, antidiabetic, antioxidant and antibacterial activity were identified from the BIOPEP prediction. The presence of a peptide sequence containing proline and leucine (LAPLPL) was found in the GPH and GIPH, which can contribute to the inhibition of ACE, as well as peptides containing aromatic residues in the C-terminal (DITWT and FLSWT) that allow the donation of electrons and stabilization of free radicals. These findings suggest that these characteristics contributed to the antioxidant activity in all in vitro assays. Hydrophobic and positively charged peptides (IFDQTLGKLR) may have made it possible to inhibit the growth of the bacteria Escherichia coli. Therefore, the peptides obtained after the GI digestion of yam proteins can contribute to the prevention of infections and chronic degenerative diseases development, with an impact on human health.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO tubérculo inhame tem importância na cultura alimentar do Nordeste, além de ser considerado um alimento funcional, que apresenta proteínas com potenciais propriedades bioativas. Objetivou-se com este estudo caracterizar o perfil peptídico e realizar a bioprospecção de peptídeos bioativos obtidos da digestão gastrointestinal simulada in vitro das proteínas do inhame (Dioscorea cayennensis) cultivado no Nordeste do Brasil. Para tanto, inicialmente, a amostra obtida foi analisada quanto a composição química parcial, seguida da obtenção da farinha para o preparo do extrato e concentrado proteico de inhame (CPI), submetido à digestão simulada in vitro, por fim caracterização e perfil dos peptídeos obtidos. O extrato proteico foi submetido à precipitação isoelétrica para obtenção do CPI, que em seguida foi dialisado, congelado e liofilizado. O CPI foi submetido à digestão gastrointestinal (GI) simulada, gerando duas fases: hidrolisado fase gástrica (HFG) e hidrólise contínua até o hidrolisado fase gastrointestinal (HFGI). As amostras: CPI, HFG e HFGI foram submetidas às análises de perfil proteico/peptídico: aminoácidos totais e livres (AAL), massa molecular (MM), hidrofobicidade por cromatografia em fase reversa e eletroforese (SDS-Tricina); perfil proteico (eletroforese-2D) e perfil peptídico. Os peptídeos identificados foram submetidos à predição de bioatividade através da Database of bioactive peptides (BIOPEP) e PeptideRanker (atividade inibitória da enzima conversora de angiotensina-ECA). Para validar os resultados obtidos in silico, foram realizadas análises de determinação de atividades biológicas in vitro: antioxidante (DPPH, ABTS, ORAC e proteção dos danos ao DNA), antibacteriana e inibidora da ECA. Os resultados da eletroforese 2D evidenciaram a presença da proteína dioscorina, como sendo a proteína majoritária do inhame. O CPI apresentou 64,0 % de proteínas, contendo todos os aminoácidos essenciais com escore adequado de acordo com a FAO, exceto os aminoácidos sulfurados. Após a digestão GI do CPI, foi evidenciada aumento da liberação significativa (p <0,5) de AAL. A eficiência da digestão GI foi verificada conforme perfil de distribuição de MM, com aproximadamente 71,0 % de peptídeos com MM <2 kDa gerados no HFGI. Os peptídeos identificados, em sua maioria são pertencentes à proteína dioscorina de D. cayennensis. Peptídeos do HFG tiveram sua MM diminuida com a progressão da hidrólise GI, resultando em peptídeos de menor MM no HFGI. Ademais, identificou-se sequências peptídicas com potenciais para peptídeos bioativos, com atividade anti-hipertensiva, antidiabética, antioxidante e antibacteriana a partir da predição por BIOPEP. Verificou-se nos HFG e HFGI a presença de sequência peptídica contendo prolina e leucina (LAPLPL) que pode contribuir para a inibição da ECA, assim como peptídeos contendo resíduos aromáticos no C-terminal (DITWT e FLSWT) que possibilitam a doação de elétrons e estabilização de radicais livres. Esses achados sugerem que essas características dos peptídeos contribuíram para a atividade antioxidante em todos os ensaios in vitro. Os peptídeos hidrofóbicos e com carga positiva (IFDQTLGKLR) podem ter possibilitado a inibição do crescimento da bactéria Escherichia coli. Portanto, os peptídeos obtidos após a digestão GI das proteínas do inhame podem contribuir na prevenção do desenvolvimento de infecções e doenças crônicas degenerativas, com impacto para a saúde humana.Universidade Federal da ParaíbaBrasilEngenharia de AlimentosPrograma de Pós-Graduação em Ciência e Tecnologia de AlimentosUFPBGadelha, Carlos Alberto de Almeidahttp://lattes.cnpq.br/0216519915668667Pacheco, Maria Teresa Bertoldohttp://lattes.cnpq.br/2123369126592009Nascimento, Edilza Silva do2021-08-11T18:28:05Z2021-02-082021-08-11T18:28:05Z2020-03-31info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/20675porhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2022-08-10T10:57:42Zoai:repositorio.ufpb.br:123456789/20675Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2022-08-10T10:57:42Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
title |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
spellingShingle |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro Nascimento, Edilza Silva do Proteína vegetal Peptídeos antibacterianos Peptídeos antioxidantes Peptídeos anti-hipertensivos BIOPEP Tuber protein Antibacterial peptides Antioxidant peptides Antihypertensive peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
title_short |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
title_full |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
title_fullStr |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
title_full_unstemmed |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
title_sort |
Inhame (Dioscorea cayennensis) cultivado na Paraíba: caracterização e produção de concentrado proteico para prospecção de peptídeos bioativos obtidos através de digestão simulada in vitro |
author |
Nascimento, Edilza Silva do |
author_facet |
Nascimento, Edilza Silva do |
author_role |
author |
dc.contributor.none.fl_str_mv |
Gadelha, Carlos Alberto de Almeida http://lattes.cnpq.br/0216519915668667 Pacheco, Maria Teresa Bertoldo http://lattes.cnpq.br/2123369126592009 |
dc.contributor.author.fl_str_mv |
Nascimento, Edilza Silva do |
dc.subject.por.fl_str_mv |
Proteína vegetal Peptídeos antibacterianos Peptídeos antioxidantes Peptídeos anti-hipertensivos BIOPEP Tuber protein Antibacterial peptides Antioxidant peptides Antihypertensive peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
topic |
Proteína vegetal Peptídeos antibacterianos Peptídeos antioxidantes Peptídeos anti-hipertensivos BIOPEP Tuber protein Antibacterial peptides Antioxidant peptides Antihypertensive peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
description |
The yam is an important tuber in Brazil Northeast food culture, also being considered a functional food, as it has proteins with potential bioactive properties. The objective of this study was to characterize the peptide profile and perform the bioprospection of bioactive peptides obtained from in vitro simulated gastrointestinal digestion of yam proteins (Dioscorea cayennensis) grown in Brazil. Therefore, the sample obtained was initially analyzed for partial chemical composition, followed by obtaining the flour for the yam extract and protein concentrate (YPC) preparation, submitted to simulated digestion in vitro, and finally the characterization and profile of the obtained peptides were performed. The protein extract was subjected to isoelectric precipitation to obtain the YPC, which was then dialyzed and freeze-dried. The YPC was submitted to simulated gastrointestinal (GI) digestion, generating two phases: the gastric phase hydrolysate (GPH) and continuous hydrolysis until the gastrointestinal phase hydrolysate (GIPH). The samples: YPC, GPH and GIPH were submitted to analysis of protein and peptide profile: total amino acids and free amino acids (FAA); molecular mass (MM); hydrophobicity by reverse phase chromatography and electrophoresis (Tricine-SDS-PAGE); protein profile (two-dimensional gel electrophoresis) and peptide profile (nano-LC-ESI-MS/MS, MALDI-TOF). The identified peptides were submitted to the bioactivity prediction through the Bioactive Peptides Database (BIOPEP) and PeptideRanker (inhibitory activity of the angiotensin-converting enzyme-ACE). To validate results obtained in silico, analysis were performed to determine biological activities in vitro: antioxidant (DPPH, ABTS, ORAC and protection from DNA damage), antibacterial and ACE’s inhibitor. The results of 2D-electrophoresis show the presence of the dioscorin protein, as the yam’s major protein. The YCP resulted in 64.0 % protein, with all essential amino acids with an appropriate score according to FAO, except sulfur amino acids. After the GI digestion of the YPC, a significant increase in the release (p <0.5) of AAL was evidenced. The efficiency of GI digestion was verified according to the distribution profile of MW, with approximately 71.0 % of peptides with MW <2 kDa generated in the GIPH. Most of the identified peptides belong to the D. cayennensis dioscorin protein. GPH peptides had their MM decreased with the progression of GI hydrolysis, resulting in smaller MM peptides in GIPH. In addition, sequences with potential for bioactive peptides, with antihypertensive, antidiabetic, antioxidant and antibacterial activity were identified from the BIOPEP prediction. The presence of a peptide sequence containing proline and leucine (LAPLPL) was found in the GPH and GIPH, which can contribute to the inhibition of ACE, as well as peptides containing aromatic residues in the C-terminal (DITWT and FLSWT) that allow the donation of electrons and stabilization of free radicals. These findings suggest that these characteristics contributed to the antioxidant activity in all in vitro assays. Hydrophobic and positively charged peptides (IFDQTLGKLR) may have made it possible to inhibit the growth of the bacteria Escherichia coli. Therefore, the peptides obtained after the GI digestion of yam proteins can contribute to the prevention of infections and chronic degenerative diseases development, with an impact on human health. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-03-31 2021-08-11T18:28:05Z 2021-02-08 2021-08-11T18:28:05Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/20675 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/20675 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Engenharia de Alimentos Programa de Pós-Graduação em Ciência e Tecnologia de Alimentos UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Engenharia de Alimentos Programa de Pós-Graduação em Ciência e Tecnologia de Alimentos UFPB |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
instname_str |
Universidade Federal da Paraíba (UFPB) |
instacron_str |
UFPB |
institution |
UFPB |
reponame_str |
Biblioteca Digital de Teses e Dissertações da UFPB |
collection |
Biblioteca Digital de Teses e Dissertações da UFPB |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
repository.mail.fl_str_mv |
diretoria@ufpb.br|| diretoria@ufpb.br |
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1801842977957478400 |