Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino

Detalhes bibliográficos
Autor(a) principal: Barbosa, Paula Perazzo de Souza
Data de Publicação: 2019
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/123456789/18872
Resumo: Goat milk is an important protein-rich food matrix that may present important functional properties in both nutritional and pharmaceutical areas. The objective of this study was to characterize the proteins present in the casein fraction (FCN) of goat milk, to obtain bioactive peptides from the FCN through in vitro hydrolysis with enzymes pepsin and trypsin; and to test the biological potential of the FCN and its peptides. For this, goat milk was collected in three distinct postpartum periods: 4 ± 2 days (colostrum); 28 ± 2 (peak lactation); and 120 ± 2 (mature milk). Then, it was skimmed by centrifugation and subjected to isoelectric precipitation by addition of hydrochloric acid to pH 4.1. The precipitate, corresponding to the FCN, was washed with distilled water, solubilized with sodium hydroxide to determine the proteins soluble by the Bradford method, dialysed against in cellulose membrane (cut-off 14 kDa), frozen and lyophilized. Partial chemical characterization composition and SDS-PAGE of the FCNs were made, which were subsequently submitted to the nLC-ESI-MS/MS technique. The FCN of milk collected from goats with 28 ± 2 days of lactation was hydrolyzed with pepsin followed by trypsin. The FCN of mature milk was characterized by means of two-dimensional electrophoresis, regarding the amino acid profile and antinociceptive activity. Subsequently, it was hydrolyzed with pepsin and trypsin, separately. The peptide profile was identified by Tris-Tricine electrophoresis. The hydrolysates had their biological potential evaluated by means of antibacterial activity for Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella spp and Pseudomonas aeroginosa; and antioxidant by the ABTS, DPPH and FRAP methods. The results of this study show that the casein fraction of goat milk is rich in proteins and changes its protein profile during lactation according to the nutritional needs of the puppy. Some proteins found, mainly in the mature milk fraction, can expose new biotechnological approaches as anticancer potential. The FCN of mature milk shows antinociceptive activity, and essential and non-essential amino acids, with high content of glutamic acid, followed by proline. Both the hydrolyzate of the proteins present in the FCN of the milk collected from goats with 28 ± 2 days of lactation and those of the FCN of the mature milk have peptides with antimicrobial properties. FCN hydrolysates of mature milk also have antioxidant properties. Thus the goat milk proteins are an interesting opportunity for application as antimicrobial agents and natural antioxidants that can be used for therapeutic purposes.
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spelling Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprinoLeite caprinoProteínas lácteasHidrólise enzimáticaPotencial biológicoGoat milkBiological potentialMilk proteinsEnzymatic hydrolysisCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOSGoat milk is an important protein-rich food matrix that may present important functional properties in both nutritional and pharmaceutical areas. The objective of this study was to characterize the proteins present in the casein fraction (FCN) of goat milk, to obtain bioactive peptides from the FCN through in vitro hydrolysis with enzymes pepsin and trypsin; and to test the biological potential of the FCN and its peptides. For this, goat milk was collected in three distinct postpartum periods: 4 ± 2 days (colostrum); 28 ± 2 (peak lactation); and 120 ± 2 (mature milk). Then, it was skimmed by centrifugation and subjected to isoelectric precipitation by addition of hydrochloric acid to pH 4.1. The precipitate, corresponding to the FCN, was washed with distilled water, solubilized with sodium hydroxide to determine the proteins soluble by the Bradford method, dialysed against in cellulose membrane (cut-off 14 kDa), frozen and lyophilized. Partial chemical characterization composition and SDS-PAGE of the FCNs were made, which were subsequently submitted to the nLC-ESI-MS/MS technique. The FCN of milk collected from goats with 28 ± 2 days of lactation was hydrolyzed with pepsin followed by trypsin. The FCN of mature milk was characterized by means of two-dimensional electrophoresis, regarding the amino acid profile and antinociceptive activity. Subsequently, it was hydrolyzed with pepsin and trypsin, separately. The peptide profile was identified by Tris-Tricine electrophoresis. The hydrolysates had their biological potential evaluated by means of antibacterial activity for Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella spp and Pseudomonas aeroginosa; and antioxidant by the ABTS, DPPH and FRAP methods. The results of this study show that the casein fraction of goat milk is rich in proteins and changes its protein profile during lactation according to the nutritional needs of the puppy. Some proteins found, mainly in the mature milk fraction, can expose new biotechnological approaches as anticancer potential. The FCN of mature milk shows antinociceptive activity, and essential and non-essential amino acids, with high content of glutamic acid, followed by proline. Both the hydrolyzate of the proteins present in the FCN of the milk collected from goats with 28 ± 2 days of lactation and those of the FCN of the mature milk have peptides with antimicrobial properties. FCN hydrolysates of mature milk also have antioxidant properties. Thus the goat milk proteins are an interesting opportunity for application as antimicrobial agents and natural antioxidants that can be used for therapeutic purposes.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqO leite caprino é uma importante matriz alimentar rica em proteínas que pode apresentar propriedades funcionais importantes, tanto na área nutricional quanto farmacêutica. Este estudo teve como objetivo caracterizar as proteínas presentes na fração caseínica (FCN) do leite caprino, obter peptídeos bioativos da FCN através da hidrolise in vitro com enzimas pepsina e tripsina; e testar o potencial biológico da FCN e dos seus peptídeos. Para tanto, o leite caprino foi coletado em três períodos distintos pós-parto: 4 ± 2 dias (colostro); 28 ± 2 (pico de lactação); e 120 ± 2 (leite maduro). Em seguida, desnatado por centrifugação e submetido à precipitação isoelétrica mediante adição de ácido clorídrico até atingir o pH 4,1. O precipitado, correspondente à FCN, foi lavado com água destilada, solubilizado com hidróxido de sódio para determinar as proteínas solúveis pelo método de Bradford, dialisado em membrana de celulose (cut-off 14 kDa), congelado e liofilizado. Foi feita a caracterização química parcial e SDS-PAGE das FCNs que, posteriormente, foram submetidas à técnica nLC-ESI-MS/MS. A FCN do leite coletado de cabras com 28 ± 2 dias de lactação foi hidrolisada com pepsina seguida de tripsina. A FCN do leite maduro foi caracterizada por meio de eletroforese bidimensional, quanto ao perfil aminoacídico e quanto à atividade antinociceptiva. Posteriormente, foi hidrolisada com pepsina e tripsina, separadamente. O perfil peptídico foi identificado por eletroforese Tris-Tricina. Os hidrolisados tiveram seu potencial biológico avaliado por meio de atividade antibacteriana para Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella spp e Pseudomonas aeroginosa; e antioxidante através dos métodos de ABTS, DPPH e FRAP. Os resultados deste estudo mostram que a fração caseínica de leite caprino é rica em proteínas, e sofre alterações no perfil proteico ao longo da lactação, de acordo com a necessidade nutricional do filhote, sendo que algumas proteínas encontradas, principalmente na fração do leite maduro, podem expor novas abordagens biotecnológicas como potencial anticancerígeno. A FCN do leite maduro apresenta atividade antinociceptiva, e aminoácidos essenciais e não essenciais, tendo com alto teor de ácido de glutâmico, seguido de prolina. Tanto o hidrolisado das proteínas presentes na FCN do leite coletado de cabras com 28 ± 2 dias de lactação quanto os da FCN do leite maduro possuem peptídeos com propriedades antimicrobianas. Os hidrolisados da FCN do leite maduro também apresentam propriedades antioxidantes. Assim, as proteínas da FCN do leite caprino são uma oportunidade interessante para aplicação como agentes antimicrobianos e antioxidantes naturais que podem ser usados para fins terapêuticos.Universidade Federal da ParaíbaBrasilEngenharia de AlimentosPrograma de Pós-Graduação em Ciência e Tecnologia de AlimentosUFPBGadelha, Tatiane Santihttp://lattes.cnpq.br/6666089513761587Barbosa, Paula Perazzo de Souza2020-12-27T11:53:43Z2020-03-282020-12-27T11:53:43Z2019-03-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/18872porhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/embargoedAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2021-08-30T14:24:26Zoai:repositorio.ufpb.br:123456789/18872Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2021-08-30T14:24:26Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false
dc.title.none.fl_str_mv Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
title Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
spellingShingle Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
Barbosa, Paula Perazzo de Souza
Leite caprino
Proteínas lácteas
Hidrólise enzimática
Potencial biológico
Goat milk
Biological potential
Milk proteins
Enzymatic hydrolysis
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS
title_short Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
title_full Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
title_fullStr Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
title_full_unstemmed Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
title_sort Análise proteômica e potencialidades de proteínas e peptídeos da fração caseínica de leite caprino
author Barbosa, Paula Perazzo de Souza
author_facet Barbosa, Paula Perazzo de Souza
author_role author
dc.contributor.none.fl_str_mv Gadelha, Tatiane Santi
http://lattes.cnpq.br/6666089513761587
dc.contributor.author.fl_str_mv Barbosa, Paula Perazzo de Souza
dc.subject.por.fl_str_mv Leite caprino
Proteínas lácteas
Hidrólise enzimática
Potencial biológico
Goat milk
Biological potential
Milk proteins
Enzymatic hydrolysis
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS
topic Leite caprino
Proteínas lácteas
Hidrólise enzimática
Potencial biológico
Goat milk
Biological potential
Milk proteins
Enzymatic hydrolysis
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS
description Goat milk is an important protein-rich food matrix that may present important functional properties in both nutritional and pharmaceutical areas. The objective of this study was to characterize the proteins present in the casein fraction (FCN) of goat milk, to obtain bioactive peptides from the FCN through in vitro hydrolysis with enzymes pepsin and trypsin; and to test the biological potential of the FCN and its peptides. For this, goat milk was collected in three distinct postpartum periods: 4 ± 2 days (colostrum); 28 ± 2 (peak lactation); and 120 ± 2 (mature milk). Then, it was skimmed by centrifugation and subjected to isoelectric precipitation by addition of hydrochloric acid to pH 4.1. The precipitate, corresponding to the FCN, was washed with distilled water, solubilized with sodium hydroxide to determine the proteins soluble by the Bradford method, dialysed against in cellulose membrane (cut-off 14 kDa), frozen and lyophilized. Partial chemical characterization composition and SDS-PAGE of the FCNs were made, which were subsequently submitted to the nLC-ESI-MS/MS technique. The FCN of milk collected from goats with 28 ± 2 days of lactation was hydrolyzed with pepsin followed by trypsin. The FCN of mature milk was characterized by means of two-dimensional electrophoresis, regarding the amino acid profile and antinociceptive activity. Subsequently, it was hydrolyzed with pepsin and trypsin, separately. The peptide profile was identified by Tris-Tricine electrophoresis. The hydrolysates had their biological potential evaluated by means of antibacterial activity for Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella spp and Pseudomonas aeroginosa; and antioxidant by the ABTS, DPPH and FRAP methods. The results of this study show that the casein fraction of goat milk is rich in proteins and changes its protein profile during lactation according to the nutritional needs of the puppy. Some proteins found, mainly in the mature milk fraction, can expose new biotechnological approaches as anticancer potential. The FCN of mature milk shows antinociceptive activity, and essential and non-essential amino acids, with high content of glutamic acid, followed by proline. Both the hydrolyzate of the proteins present in the FCN of the milk collected from goats with 28 ± 2 days of lactation and those of the FCN of the mature milk have peptides with antimicrobial properties. FCN hydrolysates of mature milk also have antioxidant properties. Thus the goat milk proteins are an interesting opportunity for application as antimicrobial agents and natural antioxidants that can be used for therapeutic purposes.
publishDate 2019
dc.date.none.fl_str_mv 2019-03-28
2020-12-27T11:53:43Z
2020-03-28
2020-12-27T11:53:43Z
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dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
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dc.identifier.uri.fl_str_mv https://repositorio.ufpb.br/jspui/handle/123456789/18872
url https://repositorio.ufpb.br/jspui/handle/123456789/18872
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/embargoedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nd/3.0/br/
eu_rights_str_mv embargoedAccess
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
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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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