Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/20073 |
Resumo: | Whey is a milk by-product usually discarded by the industry causing pollution to the environment, and therefore alternatives are needed to provide reuse. Thus, the objective of this study was to concentrate goats' milk proteins, quantify them, identify them, evaluate their antibacterial, antiproliferative and antioxidant capacity and finally carry out the hydrolysis of the proteins with the enzymes pepsin and trypsin from a experimental planning of type 2² and to analyze the antioxidant capacity of the best treatments. The whey was obtained by isoelectric precipitation, filtered, dialysed and lyophilized, thus obtaining the concentrated protein. Protein quantification was performed for total proteins and soluble by the method of Kjeldahl and Bradford, respectively. The identification of the proteins was performed by SDS-PAGE electrophoresis and the evaluation of the antibacterial activity was performed by microdilution method against the microorganisms Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli by the method of micro dilution. Antiproliferative activity was performed against the A375 line corresponding to human malignant melanoma cells. Hydrolysates were obtained from the enzymatic hydrolysis of serum proteins using the enzymes pepsin and trypsin at different pH and time values, while the degree of hydrolysis was measured by the Ortofitaldehyde method. Those that presented the best values of degree of hydrolysis were evaluated for the protein profile through Tricine SDS-PAGE electrophoresis and were tested for antioxidant activity by the DPPH and ABTS method and compared to the antioxidant activity of the native protein. The experiments of antibacterial, antiproliferative and antioxidant activity were performed in triplicate and analyzed statistically using the analysis of variance ANOVA and the Dunnet post-test. The results showed the identification in whey from milk of a heterogeneous mixture of proteins, being possible to visualize alpha-lactalbumin, beta-lactoglobulin, lactoferrin, among others. Regarding the antibacterial activity, the minimum inhibitory concentration of the serum of the milk for each microorganism was different, obtaining values that varied in the range of 120 to 30 μg / mL of protein. In relation to the inhibition of the proliferation of cancerous cells the results point the maximum value of 66%, for 24h of exposure. With respect to the hydrolysis of these proteins, from the statistical analysis it was observed that the time and the combination between time and pH were the parameters that had the greatest effect under the degree of hydrolysis of the experiments. The maximum values obtained for hydrolysis of the whey protein are approximately 35%. For these treatments it can be observed that the peptide profile and the antioxidant activity have differences when compared to the native protein, showing that the hydrolysis can generate bioactive peptides capable of performing antioxidant activity. Finally, the results suggest that goat's milk serum is an important functional matrix that has a protein composition of great interest and applicability exerting potential biological activities and thus being able to be reused in order to obtain bioactive peptides. |
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Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisadoAtividade biológicaCaprinoSubprodutoProteômicaPeptídeosBiological activityBy-productProteomicsResiduePeptidesCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOSWhey is a milk by-product usually discarded by the industry causing pollution to the environment, and therefore alternatives are needed to provide reuse. Thus, the objective of this study was to concentrate goats' milk proteins, quantify them, identify them, evaluate their antibacterial, antiproliferative and antioxidant capacity and finally carry out the hydrolysis of the proteins with the enzymes pepsin and trypsin from a experimental planning of type 2² and to analyze the antioxidant capacity of the best treatments. The whey was obtained by isoelectric precipitation, filtered, dialysed and lyophilized, thus obtaining the concentrated protein. Protein quantification was performed for total proteins and soluble by the method of Kjeldahl and Bradford, respectively. The identification of the proteins was performed by SDS-PAGE electrophoresis and the evaluation of the antibacterial activity was performed by microdilution method against the microorganisms Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli by the method of micro dilution. Antiproliferative activity was performed against the A375 line corresponding to human malignant melanoma cells. Hydrolysates were obtained from the enzymatic hydrolysis of serum proteins using the enzymes pepsin and trypsin at different pH and time values, while the degree of hydrolysis was measured by the Ortofitaldehyde method. Those that presented the best values of degree of hydrolysis were evaluated for the protein profile through Tricine SDS-PAGE electrophoresis and were tested for antioxidant activity by the DPPH and ABTS method and compared to the antioxidant activity of the native protein. The experiments of antibacterial, antiproliferative and antioxidant activity were performed in triplicate and analyzed statistically using the analysis of variance ANOVA and the Dunnet post-test. The results showed the identification in whey from milk of a heterogeneous mixture of proteins, being possible to visualize alpha-lactalbumin, beta-lactoglobulin, lactoferrin, among others. Regarding the antibacterial activity, the minimum inhibitory concentration of the serum of the milk for each microorganism was different, obtaining values that varied in the range of 120 to 30 μg / mL of protein. In relation to the inhibition of the proliferation of cancerous cells the results point the maximum value of 66%, for 24h of exposure. With respect to the hydrolysis of these proteins, from the statistical analysis it was observed that the time and the combination between time and pH were the parameters that had the greatest effect under the degree of hydrolysis of the experiments. The maximum values obtained for hydrolysis of the whey protein are approximately 35%. For these treatments it can be observed that the peptide profile and the antioxidant activity have differences when compared to the native protein, showing that the hydrolysis can generate bioactive peptides capable of performing antioxidant activity. Finally, the results suggest that goat's milk serum is an important functional matrix that has a protein composition of great interest and applicability exerting potential biological activities and thus being able to be reused in order to obtain bioactive peptides.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqO soro do leite é um subproduto lácteo usualmente descartado pela indústria ocasionando poluição ao ambiente, sendo assim necessárias alternativas que proporcionem sua reutilização. Dessa maneira, o objetivo desse estudo foi concentrar as proteínas do soro do leite caprino, quantificar estas, identifica-las, avaliar sua capacidade antibacteriana, antiproliferativa e antioxidante e por fim realizar a hidrólise das mesmas com as enzimas pepsina e tripsina a partir de um planejamento experimental fatorial do tipo 2² e analisar a capacidade antioxidante dos melhores tratamentos. O soro do leite foi obtido por precipitação isoelétrica, filtrado, dialisado e liofilizado obtendo-se assim a proteína concentrada. A quantificação das proteínas foi realizada para proteínas totais e solúveis pelo método de Kjeldahl e Bradford, respectivamente. A identificação das proteínas foi realizada por eletroforese SDS-PAGE e a avaliação da atividade antibacteriana foi realizada pelo método de microdiluição frente aos micro-organismos Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Pseudomonas aeruginosa e Escherichia coli pelo método de microdiluição. A atividade antiproliferativa foi realizada frente à linhagem A375 correspondente a células de melanoma maligno humano. Os hidrolisados foram obtidos a partir da hidrolise enzimática das proteínas do soro utilizando as enzimas pepsina e tripsina em diferentes valores de pH e tempo, enquanto o grau de hidrólise foi medido através do método de Ortofitaldeído. Aqueles que apresentaram os melhores valores de grau de hidrólise foram avaliados quanto ao perfil proteico através de eletroforese Tricina SDS-PAGE e foram testados quanto à atividade antioxidante pelo método DPPH e ABTS e comparados quanto a atividade antioxidante da proteína nativa. Os experimentos de atividade antibacteriana, antiproliferativa e antioxidante foram realizados em triplicata e analisados estatisticamente utilizando a análise de variância ANOVA e o pós-teste de Dunnet. Os resultados mostraram a identificação no soro do leite concentrado de uma mistura heterogênea de proteínas, sendo possível visualizar alfa-lactoalbumina, beta-lactoglobulina, lactoferrina, dentre outras. Com relação à atividade antibacteriana a concentração inibitória miníma do soro do leite para cada microrganismo foi diferente, obtendo-se valores que variaram na faixa de 120 a 30µg/mL de proteína. Em relação a inibição da proliferação de células cancerígenas os resultados apontam o valor máximo de 66%, para 24h de exposição. Com relação a hidrólise destas proteínas, a partir da análise estatística foi observado que o tempo e a combinação entre tempo e pH foram os parâmetros que exerceram maior efeito sob o grau de hidrólise dos experimentos. Os valores máximos obtidos de hidrólise da proteína do soro são de aproximadamente 35%. Para estes tratamentos pode se observar que o perfil peptídico e a atividade antioxidante possuem diferenças quando comparado à proteína nativa mostrando que a hidrólise pode gerar peptídeos bioativos capazes de desempenhar atividade antioxidante Por fim, os resultados sugerem que o soro do leite de cabra é uma importante matriz funcional que possui uma composição proteica de bastante interesse e aplicabilidade exercendo potenciais atividades biológicas e assim podendo ser reaproveitado a fim de obter-se peptídeos bioativos.Universidade Federal da ParaíbaBrasilEngenharia de AlimentosPrograma de Pós-Graduação em Ciência e Tecnologia de AlimentosUFPBGadelha, Tatiane Santihttp://lattes.cnpq.br/6666089513761587Campos, Maria Isabel Ferreira2021-05-21T18:17:59Z2019-04-242021-05-21T18:17:59Z2018-04-10info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/20073porhttp://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:UFPB2021-06-11T12:48:41Zoai:repositorio.ufpb.br:123456789/20073Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2021-06-11T12:48:41Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
title |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
spellingShingle |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado Campos, Maria Isabel Ferreira Atividade biológica Caprino Subproduto Proteômica Peptídeos Biological activity By-product Proteomics Residue Peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
title_short |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
title_full |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
title_fullStr |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
title_full_unstemmed |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
title_sort |
Identificação do perfil protéico e bioprospecção do soro do leite de cabra concentrado e hidrolisado |
author |
Campos, Maria Isabel Ferreira |
author_facet |
Campos, Maria Isabel Ferreira |
author_role |
author |
dc.contributor.none.fl_str_mv |
Gadelha, Tatiane Santi http://lattes.cnpq.br/6666089513761587 |
dc.contributor.author.fl_str_mv |
Campos, Maria Isabel Ferreira |
dc.subject.por.fl_str_mv |
Atividade biológica Caprino Subproduto Proteômica Peptídeos Biological activity By-product Proteomics Residue Peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
topic |
Atividade biológica Caprino Subproduto Proteômica Peptídeos Biological activity By-product Proteomics Residue Peptides CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
description |
Whey is a milk by-product usually discarded by the industry causing pollution to the environment, and therefore alternatives are needed to provide reuse. Thus, the objective of this study was to concentrate goats' milk proteins, quantify them, identify them, evaluate their antibacterial, antiproliferative and antioxidant capacity and finally carry out the hydrolysis of the proteins with the enzymes pepsin and trypsin from a experimental planning of type 2² and to analyze the antioxidant capacity of the best treatments. The whey was obtained by isoelectric precipitation, filtered, dialysed and lyophilized, thus obtaining the concentrated protein. Protein quantification was performed for total proteins and soluble by the method of Kjeldahl and Bradford, respectively. The identification of the proteins was performed by SDS-PAGE electrophoresis and the evaluation of the antibacterial activity was performed by microdilution method against the microorganisms Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli by the method of micro dilution. Antiproliferative activity was performed against the A375 line corresponding to human malignant melanoma cells. Hydrolysates were obtained from the enzymatic hydrolysis of serum proteins using the enzymes pepsin and trypsin at different pH and time values, while the degree of hydrolysis was measured by the Ortofitaldehyde method. Those that presented the best values of degree of hydrolysis were evaluated for the protein profile through Tricine SDS-PAGE electrophoresis and were tested for antioxidant activity by the DPPH and ABTS method and compared to the antioxidant activity of the native protein. The experiments of antibacterial, antiproliferative and antioxidant activity were performed in triplicate and analyzed statistically using the analysis of variance ANOVA and the Dunnet post-test. The results showed the identification in whey from milk of a heterogeneous mixture of proteins, being possible to visualize alpha-lactalbumin, beta-lactoglobulin, lactoferrin, among others. Regarding the antibacterial activity, the minimum inhibitory concentration of the serum of the milk for each microorganism was different, obtaining values that varied in the range of 120 to 30 μg / mL of protein. In relation to the inhibition of the proliferation of cancerous cells the results point the maximum value of 66%, for 24h of exposure. With respect to the hydrolysis of these proteins, from the statistical analysis it was observed that the time and the combination between time and pH were the parameters that had the greatest effect under the degree of hydrolysis of the experiments. The maximum values obtained for hydrolysis of the whey protein are approximately 35%. For these treatments it can be observed that the peptide profile and the antioxidant activity have differences when compared to the native protein, showing that the hydrolysis can generate bioactive peptides capable of performing antioxidant activity. Finally, the results suggest that goat's milk serum is an important functional matrix that has a protein composition of great interest and applicability exerting potential biological activities and thus being able to be reused in order to obtain bioactive peptides. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-04-10 2019-04-24 2021-05-21T18:17:59Z 2021-05-21T18:17:59Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/20073 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/20073 |
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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1801842973815603200 |