Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano

Detalhes bibliográficos
Autor(a) principal: Favero, Adriano
Data de Publicação: 2019
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações do UNIOESTE
Texto Completo: http://tede.unioeste.br/handle/tede/4673
Resumo: Lectins are proteins of non-immune origin, which arouse biotechnological interest in the health area, due to their selective and reversible carbohydrate binding capacity, mediating cellular recognition processes. The objective of this work was to extract lectins, determine their specificity to carbohydrates and evaluate the antibacterial potential in the extract and precipitated and dialysated fractions of the seeds of Eugenia pyriformis. The aqueous extraction was performed in phosphate Buffer pH 7.0, 50 mM with NaCl 0.15 M, followed by fractionation with ammonium sulfate in the saturations of 0-40 (FPD 0-40) and 40-80% (FPD 40-80). The phenolic and proteic dosages were performed by the FolinCicalteau and Bradford methods, respectively. For the detection of lectins, the native and thermally denatured samples were subjected to hemagglutinating activity (HA) tests in Human erythrocytes (ABO), by the serial dilution method. The inhibition of the HA of the lectins was rehearsed with the pre-incubation of the native samples with the carbohydrates D (+)-glucose, D(+)galactose, D(+)glucose, D(+)maltose, D(+)mannose, D(+) sucrose, D(+) ribose, D(+) fucose, D(+) arabinose, D(+) glucosamina, D(+) lactose monohydrate , D(+) galactosamine, N-Acetyl-D-glucosamine, D(+) Trehalose dihydrate, D(+) rhamnose e D-(+)-raffinose pentahydrate. The antibacterial assays were performed by means of disc diffusion for the strains: Staphylococcus aureus, Geobacillus stearothermophilus, Pseudomonas aeruginosa, Shigella flexneri, Escherichia coli, Klebsiella sp., Morganella Sp., Serratia sp., Proteus Mirabilis, Enterococcus faecalis, Bacillus cereus, Streptococcus uberis, Streptococcus pyogenes and Streptococcus agalactiae. The dosages revealed that the seeds contain low protein content (1%) and phenols (0.1%). The HA assays of the native samples resulted positive for ABO erythrocytes, with preference for type A and higher HA (1024) for FPD 40-80. After the thermal denaturation of the samples, the activities suffered a considerable decrease, showing that most of the HA is due to the presence of lectins, which presented affinity mainly for D (+)-galactose, D (+)-arabinose, D (+)- lactose monohydrate and D (+)- galactosamine. The antibacterial tests of the native samples obtained formation of inhibition halos for the strains Staphylococcus aureus, Streptococcus uberis, Enterococcus faecalis, Shigella flexneri, Escherichia coli, Klebsiella sp. and Serratia sp., for all tested fractions. The loss of antibacterial activity in denatures samples shows the proteic nature of the compounds responsible for the effect. The diversity of the specificity of carbohydrates in each sample suggests the presence of at least two lectins in the seeds.
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spelling Lucca, Rosemeire Aparecida da Silva dehttp://lattes.cnpq.br/7573562459264964Lucca, Rosemeire Aparecida da Silva dehttp://lattes.cnpq.br/7573562459264964Follador, Franciele Ani Caovillahttp://lattes.cnpq.br/9329326606646660Moura, Alexandre Carvalho dehttp://lattes.cnpq.br/1811737564247729http://lattes.cnpq.br/9971263606030846Favero, Adriano2020-02-10T11:19:15Z2019-09-26FAVERO, Adriano. Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano. 2019. 83 f. Dissertação( Mestrado em Ciências Aplicadas à Saúde) - Universidade Estadual do Oeste do Paraná, Francisco Beltrão, 2019.http://tede.unioeste.br/handle/tede/4673Lectins are proteins of non-immune origin, which arouse biotechnological interest in the health area, due to their selective and reversible carbohydrate binding capacity, mediating cellular recognition processes. The objective of this work was to extract lectins, determine their specificity to carbohydrates and evaluate the antibacterial potential in the extract and precipitated and dialysated fractions of the seeds of Eugenia pyriformis. The aqueous extraction was performed in phosphate Buffer pH 7.0, 50 mM with NaCl 0.15 M, followed by fractionation with ammonium sulfate in the saturations of 0-40 (FPD 0-40) and 40-80% (FPD 40-80). The phenolic and proteic dosages were performed by the FolinCicalteau and Bradford methods, respectively. For the detection of lectins, the native and thermally denatured samples were subjected to hemagglutinating activity (HA) tests in Human erythrocytes (ABO), by the serial dilution method. The inhibition of the HA of the lectins was rehearsed with the pre-incubation of the native samples with the carbohydrates D (+)-glucose, D(+)galactose, D(+)glucose, D(+)maltose, D(+)mannose, D(+) sucrose, D(+) ribose, D(+) fucose, D(+) arabinose, D(+) glucosamina, D(+) lactose monohydrate , D(+) galactosamine, N-Acetyl-D-glucosamine, D(+) Trehalose dihydrate, D(+) rhamnose e D-(+)-raffinose pentahydrate. The antibacterial assays were performed by means of disc diffusion for the strains: Staphylococcus aureus, Geobacillus stearothermophilus, Pseudomonas aeruginosa, Shigella flexneri, Escherichia coli, Klebsiella sp., Morganella Sp., Serratia sp., Proteus Mirabilis, Enterococcus faecalis, Bacillus cereus, Streptococcus uberis, Streptococcus pyogenes and Streptococcus agalactiae. The dosages revealed that the seeds contain low protein content (1%) and phenols (0.1%). The HA assays of the native samples resulted positive for ABO erythrocytes, with preference for type A and higher HA (1024) for FPD 40-80. After the thermal denaturation of the samples, the activities suffered a considerable decrease, showing that most of the HA is due to the presence of lectins, which presented affinity mainly for D (+)-galactose, D (+)-arabinose, D (+)- lactose monohydrate and D (+)- galactosamine. The antibacterial tests of the native samples obtained formation of inhibition halos for the strains Staphylococcus aureus, Streptococcus uberis, Enterococcus faecalis, Shigella flexneri, Escherichia coli, Klebsiella sp. and Serratia sp., for all tested fractions. The loss of antibacterial activity in denatures samples shows the proteic nature of the compounds responsible for the effect. The diversity of the specificity of carbohydrates in each sample suggests the presence of at least two lectins in the seeds.Lectinas são proteínas de origem não imune, que despertam interesse biotecnológico na área da saúde, devido à sua capacidade de ligação seletiva e reversível aos carboidratos, mediando processos de reconhecimento celular. O objetivo deste trabalho foi extrair lectinas, determinar sua especificidade aos carboidratos e avaliar o potencial antibacteriano no extrato e frações precipitadas e dialisadas das sementes de Eugenia pyriformis. A extração aquosa foi realizada em tampão fosfato pH 7,0, 50 mM com NaCl 0,15 M, seguida de fracionamento com sulfato de amônio nas saturações de 040 (FPD 0-40) e 40-80% (FPD 40-80). As dosagens fenólicas e proteicas foram realizadas pelos métodos de Folin-Cicalteau e Bradford, respectivamente. Para a detecção das lectinas, as amostras nativas e termicamente desnaturadas foram submetidas a testes de atividade hemaglutinante (AH) em eritrócitos humanos (ABO), pelo método de diluição seriada. A inibição da AH das lectinas foi ensaiada com a pré-incubação das amostras nativas com os carboidratos D(+)galactose, D(+)glicose, D(+)maltose, D(+)manose, D(+) sacarose, D(+) ribose, D(+) fucose, D(+) arabinose, D(+) glicosamina, D(+) lactose monohidratada, D(+) galactosamina, N-acetil D-glicosamina, D(+) trealose di-hidratada, D(+) ramnose e D-(+)-raffinose penta-hidratada. Os ensaios antibacterianos foram realizados por meio de disco-difusão para as cepas: Staphylococcus aureus, Geobacillus stearothermophilus, Pseudomonas aeruginosa, Shigella flexneri, Escherichia coli, Klebsiella sp., Morganella sp., Serratia sp., Proteus Mirabilis, Enterococcus faecalis, Bacillus cereus, Streptococcus uberis, Streptococcus pyogenes e Streptococcus agalactiae. As dosagens revelaram que as sementes contêm baixo teor de proteínas (1%) e fenóis (0,1%). Os ensaios de AH das amostras nativas resultaram positivos para hemácias ABO, com preferência para o tipo A e maior AH (1024) para a FPD 40-80. Após a desnaturação térmica das amostras, as atividades sofreram considerável diminuição (de 1024 para 16), mostrando que a maior parte da AH é devida à presença de lectinas, que apresentaram afinidade principalmente por D (+) -galactose, D (+) -arabinose, D(+)lactose e D(+)-galactosamina. Os testes antibacterianos das amostras nativas apresentaram formação de halos de inibição para as cepas Staphylococcus aureus, Streptococcus uberis, Enterococcus faecalis, Shigella flexneri, Escherichia coli, Klebsiella sp. e Serratia sp., para todas as frações testadas. A perda da atividade antibacteriana nas amostras desnaturadas mostra a natureza proteica dos compostos responsáveis pelo efeito. A diversidade da especificidade dos carboidratos em cada amostra sugere a presença de no mínimo duas lectinas nas sementes.Submitted by Almir Squinsani (almir.squinsani@unioeste.br) on 2020-02-10T11:19:15Z No. of bitstreams: 2 Adriano_Favero_2019.pdf: 1556553 bytes, checksum: 6198503cced6b15067a27aba3807563c (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2020-02-10T11:19:15Z (GMT). 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dc.title.por.fl_str_mv Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
dc.title.alternative.eng.fl_str_mv Isolation, characterization of the lectin of Eugenia pyriformis seeds and antimicrobial potential
title Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
spellingShingle Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
Favero, Adriano
Antibacteriano
Antifúngico
Hemaglutinação
Myrtacea
Uvaia
Antibacterial,
Antifungal
Hemagglutination
Myrtacea
Uvaia
CIÊNCIAS DA SAÚDE
title_short Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
title_full Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
title_fullStr Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
title_full_unstemmed Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
title_sort Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano
author Favero, Adriano
author_facet Favero, Adriano
author_role author
dc.contributor.advisor1.fl_str_mv Lucca, Rosemeire Aparecida da Silva de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7573562459264964
dc.contributor.referee1.fl_str_mv Lucca, Rosemeire Aparecida da Silva de
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/7573562459264964
dc.contributor.referee2.fl_str_mv Follador, Franciele Ani Caovilla
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/9329326606646660
dc.contributor.referee3.fl_str_mv Moura, Alexandre Carvalho de
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/1811737564247729
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9971263606030846
dc.contributor.author.fl_str_mv Favero, Adriano
contributor_str_mv Lucca, Rosemeire Aparecida da Silva de
Lucca, Rosemeire Aparecida da Silva de
Follador, Franciele Ani Caovilla
Moura, Alexandre Carvalho de
dc.subject.por.fl_str_mv Antibacteriano
Antifúngico
Hemaglutinação
Myrtacea
Uvaia
topic Antibacteriano
Antifúngico
Hemaglutinação
Myrtacea
Uvaia
Antibacterial,
Antifungal
Hemagglutination
Myrtacea
Uvaia
CIÊNCIAS DA SAÚDE
dc.subject.eng.fl_str_mv Antibacterial,
Antifungal
Hemagglutination
Myrtacea
Uvaia
dc.subject.cnpq.fl_str_mv CIÊNCIAS DA SAÚDE
description Lectins are proteins of non-immune origin, which arouse biotechnological interest in the health area, due to their selective and reversible carbohydrate binding capacity, mediating cellular recognition processes. The objective of this work was to extract lectins, determine their specificity to carbohydrates and evaluate the antibacterial potential in the extract and precipitated and dialysated fractions of the seeds of Eugenia pyriformis. The aqueous extraction was performed in phosphate Buffer pH 7.0, 50 mM with NaCl 0.15 M, followed by fractionation with ammonium sulfate in the saturations of 0-40 (FPD 0-40) and 40-80% (FPD 40-80). The phenolic and proteic dosages were performed by the FolinCicalteau and Bradford methods, respectively. For the detection of lectins, the native and thermally denatured samples were subjected to hemagglutinating activity (HA) tests in Human erythrocytes (ABO), by the serial dilution method. The inhibition of the HA of the lectins was rehearsed with the pre-incubation of the native samples with the carbohydrates D (+)-glucose, D(+)galactose, D(+)glucose, D(+)maltose, D(+)mannose, D(+) sucrose, D(+) ribose, D(+) fucose, D(+) arabinose, D(+) glucosamina, D(+) lactose monohydrate , D(+) galactosamine, N-Acetyl-D-glucosamine, D(+) Trehalose dihydrate, D(+) rhamnose e D-(+)-raffinose pentahydrate. The antibacterial assays were performed by means of disc diffusion for the strains: Staphylococcus aureus, Geobacillus stearothermophilus, Pseudomonas aeruginosa, Shigella flexneri, Escherichia coli, Klebsiella sp., Morganella Sp., Serratia sp., Proteus Mirabilis, Enterococcus faecalis, Bacillus cereus, Streptococcus uberis, Streptococcus pyogenes and Streptococcus agalactiae. The dosages revealed that the seeds contain low protein content (1%) and phenols (0.1%). The HA assays of the native samples resulted positive for ABO erythrocytes, with preference for type A and higher HA (1024) for FPD 40-80. After the thermal denaturation of the samples, the activities suffered a considerable decrease, showing that most of the HA is due to the presence of lectins, which presented affinity mainly for D (+)-galactose, D (+)-arabinose, D (+)- lactose monohydrate and D (+)- galactosamine. The antibacterial tests of the native samples obtained formation of inhibition halos for the strains Staphylococcus aureus, Streptococcus uberis, Enterococcus faecalis, Shigella flexneri, Escherichia coli, Klebsiella sp. and Serratia sp., for all tested fractions. The loss of antibacterial activity in denatures samples shows the proteic nature of the compounds responsible for the effect. The diversity of the specificity of carbohydrates in each sample suggests the presence of at least two lectins in the seeds.
publishDate 2019
dc.date.issued.fl_str_mv 2019-09-26
dc.date.accessioned.fl_str_mv 2020-02-10T11:19:15Z
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.citation.fl_str_mv FAVERO, Adriano. Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano. 2019. 83 f. Dissertação( Mestrado em Ciências Aplicadas à Saúde) - Universidade Estadual do Oeste do Paraná, Francisco Beltrão, 2019.
dc.identifier.uri.fl_str_mv http://tede.unioeste.br/handle/tede/4673
identifier_str_mv FAVERO, Adriano. Isolamento, caracterização da lectina das sementes de Eugenia pyriformis e potencial antimicrobiano. 2019. 83 f. Dissertação( Mestrado em Ciências Aplicadas à Saúde) - Universidade Estadual do Oeste do Paraná, Francisco Beltrão, 2019.
url http://tede.unioeste.br/handle/tede/4673
dc.language.iso.fl_str_mv por
language por
dc.relation.program.fl_str_mv 6290525253230630664
dc.relation.confidence.fl_str_mv 600
600
dc.relation.department.fl_str_mv 292441653440865123
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual do Oeste do Paraná
Francisco Beltrão
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Ciências Aplicadas à Saúde
dc.publisher.initials.fl_str_mv UNIOESTE
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Estadual do Oeste do Paraná
Francisco Beltrão
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações do UNIOESTE
instname:Universidade Estadual do Oeste do Paraná (UNIOESTE)
instacron:UNIOESTE
instname_str Universidade Estadual do Oeste do Paraná (UNIOESTE)
instacron_str UNIOESTE
institution UNIOESTE
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