Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/15134 |
Resumo: | In this work was studied the immobilization process β-galactosidase from Aspergillus oryzae by adsorption and cross-linking with glutaraldehyde, using as carrier ions-exchangers such as Duolite A-568, Duolite S-761, Dowex Marathon A, Dowex Marathon C and Amberlite 252 Na. Through innitial results, Duolite A-568 was choosed for the continuance of the work. The influence of the enzyme concentration and pH in the immobilization process was studied using a Central Composit Design (CCD) for a fixed time of 12 hours and temperature of 25°C. The optimal conditions for enzyme immobilization were pH 4,5 and galactosidase concentration 16 g/L. In the sequence, was studied the influence of glutaraldehyde concentration as cross-linking reagent and reaction time in the activity and in stability of the immobilized biocalyst, which were of 3,5 g/L and 1,50 h. The residual activity of the immobilized enzyme without cross-linking with glutaraldehyde after 30 uses was 51% as compared with the initial activity, while the enzyme immobilized with cross-linking was 90%. The immobilized enzyme with cross-linking presented higher pH stabilyt pH when compared to that without the referred treatment. The simultaneous influence of pH and temperature on the immobilized enzyme activity was studied through a PCC with the biocalyst produced in the optimized conditions of immobilization process. With the technique response surface it was possible to obtain the optimized pH of 4,1 and temperature of 34°C. The influence of the lactose concentration was studied in the range of 5 to 140 g/L and the Michaelis-Menten model was adjusted to the experimental results, with values of Vm and Km of 0,71 U and 12.07 g/L, respectively. In the study of the influence of the galactose as inhibitor of the lactose hydrolysis, the competitive inhibition model was adjusted to the experimental results and the values of Vm, Km and Ki were 0,77 U, 12,07 g/L and 4,94 g/L respectively. |
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2016-06-22T18:41:38Z2010-03-022016-06-22T18:41:38Z2009-08-28GUIDINI, Carla Zanella. Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica. 2009. 136 f. Dissertação (Mestrado em Engenharias) - Universidade Federal de Uberlândia, Uberlândia, 2009.https://repositorio.ufu.br/handle/123456789/15134In this work was studied the immobilization process β-galactosidase from Aspergillus oryzae by adsorption and cross-linking with glutaraldehyde, using as carrier ions-exchangers such as Duolite A-568, Duolite S-761, Dowex Marathon A, Dowex Marathon C and Amberlite 252 Na. Through innitial results, Duolite A-568 was choosed for the continuance of the work. The influence of the enzyme concentration and pH in the immobilization process was studied using a Central Composit Design (CCD) for a fixed time of 12 hours and temperature of 25°C. The optimal conditions for enzyme immobilization were pH 4,5 and galactosidase concentration 16 g/L. In the sequence, was studied the influence of glutaraldehyde concentration as cross-linking reagent and reaction time in the activity and in stability of the immobilized biocalyst, which were of 3,5 g/L and 1,50 h. The residual activity of the immobilized enzyme without cross-linking with glutaraldehyde after 30 uses was 51% as compared with the initial activity, while the enzyme immobilized with cross-linking was 90%. The immobilized enzyme with cross-linking presented higher pH stabilyt pH when compared to that without the referred treatment. The simultaneous influence of pH and temperature on the immobilized enzyme activity was studied through a PCC with the biocalyst produced in the optimized conditions of immobilization process. With the technique response surface it was possible to obtain the optimized pH of 4,1 and temperature of 34°C. The influence of the lactose concentration was studied in the range of 5 to 140 g/L and the Michaelis-Menten model was adjusted to the experimental results, with values of Vm and Km of 0,71 U and 12.07 g/L, respectively. In the study of the influence of the galactose as inhibitor of the lactose hydrolysis, the competitive inhibition model was adjusted to the experimental results and the values of Vm, Km and Ki were 0,77 U, 12,07 g/L and 4,94 g/L respectively.Neste trabalho estudou-se o processo de imobilização da enzima β-galactosidase de Aspergillus oryzae por adsorção e ligação cruzada com glutaraldeído, utilizando como suporte resinas de troca iônica Duolite A-568, Duolite S-761, Dowex Marathon A, Dowex Marathon C e Amberlite 252 Na. Resultados preliminares da resina Duolite A-568 apresentaram melhor retenção de atividade enzimática, sendo assim a mesma foi utilizada para os trabalhos subsequentes. A influência da concentração da enzima e do pH no processo de imobilização foi estudada empregando um Planejamento Composto Central (PCC) fixando tempo de 12 horas e temperatura de 25°C. As condições ótimas de imobilização foram pH 4,5 e concentração de β-galactosidase de 16 g/L. Na seqüência, foi realizado um estudo da influência da concentração de glutaraldeído como agente reticulante e do tempo de reação na atividade e na estabilidade do biocatalisador imobilizado. A concentração de glutaraldeído que implicou em um biocatalisador de maior atividade enzimática foi de 3,5 g/L, num tempo de reação de 1,50 horas. A atividade da enzima imobilizada sem tratamento com glutaraldeído após 30 usos foi de 51% em relação à inicial, enquanto que a da enzima imobilizada com reticulação foi de 90%. A enzima imobilizada empregando ligação cruzada com glutaraldeído foi muito mais estável em relação ao pH quando comparada àquela sem o referido tratamento. A seguir foi realizado um estudo da influência conjunta da temperatura e do pH na atividade da enzima imbilizada por meio de um PCC, utilizando o biocatalisador produzido nas condições otimizadas de imobilização e ligação cruzada. Com o emprego da técnica de superfície de resposta foi possível otimizar, dentro das faixas estudadas, as variáveis pH igual a 4,1 e temperatura de 34°C. A influência da concentração de lactose foi estudada para uma faixa de 5 a 140 g/L e o modelo de Michaelis-Menten ajustou-se bem aos resultados experimentais, com valores de Vm e Km de 0,71 U e 12,07 g/L, respectivamente. No estudo da influência da galactose como inibidor da reação de hidrólise, o modelo que melhor se ajustou aos resultados experimentais foi o modelo de inibição competitivo, com valores de Vm, Km e Ki iguais 0,77 U, 12,07 g/L e 4,94 g/L respectivamente.Mestre em Engenharia Químicaapplication/pdfporUniversidade Federal de UberlândiaPrograma de Pós-graduação em Engenharia QuímicaUFUBREngenhariasgalactosidaseHidrólise da lactoseImobilizaçãoDuolite A-568GlutaraldeídoHidróliseLactose hydrolysisImmobilizationDuolite A-568GlutaraldehydeCNPQ::ENGENHARIAS::ENGENHARIA QUIMICAImobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônicainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisCardoso, Vicelma Luizhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787074J7Ribeiro, Eloízio Júliohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721952Y1Coutinho Filho, Ubirajarahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797915J2Resende, Miriam Maria dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703538D3Servulo, Eliana Flavia Camporesehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797632Y9http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4212184E2Guidini, Carla Zanella81756759info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFUTHUMBNAILcarla.pdf.jpgcarla.pdf.jpgGenerated Thumbnailimage/jpeg1368https://repositorio.ufu.br/bitstream/123456789/15134/3/carla.pdf.jpg13d16b1ebca40a2861faff42abbd7668MD53ORIGINALcarla.pdfapplication/pdf2903087https://repositorio.ufu.br/bitstream/123456789/15134/1/carla.pdfa7bddf585a85bf245d6d0e90cf17f0f8MD51TEXTcarla.pdf.txtcarla.pdf.txtExtracted texttext/plain243837https://repositorio.ufu.br/bitstream/123456789/15134/2/carla.pdf.txt02491ede73bdf93b9b5878c266f62695MD52123456789/151342021-09-29 13:57:18.156oai:repositorio.ufu.br:123456789/15134Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2021-09-29T16:57:18Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.por.fl_str_mv |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
title |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
spellingShingle |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica Guidini, Carla Zanella galactosidase Hidrólise da lactose Imobilização Duolite A-568 Glutaraldeído Hidrólise Lactose hydrolysis Immobilization Duolite A-568 Glutaraldehyde CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA |
title_short |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
title_full |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
title_fullStr |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
title_full_unstemmed |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
title_sort |
Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica |
author |
Guidini, Carla Zanella |
author_facet |
Guidini, Carla Zanella |
author_role |
author |
dc.contributor.advisor-co1.fl_str_mv |
Cardoso, Vicelma Luiz |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787074J7 |
dc.contributor.advisor1.fl_str_mv |
Ribeiro, Eloízio Júlio |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721952Y1 |
dc.contributor.referee1.fl_str_mv |
Coutinho Filho, Ubirajara |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797915J2 |
dc.contributor.referee2.fl_str_mv |
Resende, Miriam Maria de |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703538D3 |
dc.contributor.referee3.fl_str_mv |
Servulo, Eliana Flavia Camporese |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797632Y9 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4212184E2 |
dc.contributor.author.fl_str_mv |
Guidini, Carla Zanella |
contributor_str_mv |
Cardoso, Vicelma Luiz Ribeiro, Eloízio Júlio Coutinho Filho, Ubirajara Resende, Miriam Maria de Servulo, Eliana Flavia Camporese |
dc.subject.por.fl_str_mv |
galactosidase Hidrólise da lactose Imobilização Duolite A-568 Glutaraldeído Hidrólise |
topic |
galactosidase Hidrólise da lactose Imobilização Duolite A-568 Glutaraldeído Hidrólise Lactose hydrolysis Immobilization Duolite A-568 Glutaraldehyde CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA |
dc.subject.eng.fl_str_mv |
Lactose hydrolysis Immobilization Duolite A-568 Glutaraldehyde |
dc.subject.cnpq.fl_str_mv |
CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA |
description |
In this work was studied the immobilization process β-galactosidase from Aspergillus oryzae by adsorption and cross-linking with glutaraldehyde, using as carrier ions-exchangers such as Duolite A-568, Duolite S-761, Dowex Marathon A, Dowex Marathon C and Amberlite 252 Na. Through innitial results, Duolite A-568 was choosed for the continuance of the work. The influence of the enzyme concentration and pH in the immobilization process was studied using a Central Composit Design (CCD) for a fixed time of 12 hours and temperature of 25°C. The optimal conditions for enzyme immobilization were pH 4,5 and galactosidase concentration 16 g/L. In the sequence, was studied the influence of glutaraldehyde concentration as cross-linking reagent and reaction time in the activity and in stability of the immobilized biocalyst, which were of 3,5 g/L and 1,50 h. The residual activity of the immobilized enzyme without cross-linking with glutaraldehyde after 30 uses was 51% as compared with the initial activity, while the enzyme immobilized with cross-linking was 90%. The immobilized enzyme with cross-linking presented higher pH stabilyt pH when compared to that without the referred treatment. The simultaneous influence of pH and temperature on the immobilized enzyme activity was studied through a PCC with the biocalyst produced in the optimized conditions of immobilization process. With the technique response surface it was possible to obtain the optimized pH of 4,1 and temperature of 34°C. The influence of the lactose concentration was studied in the range of 5 to 140 g/L and the Michaelis-Menten model was adjusted to the experimental results, with values of Vm and Km of 0,71 U and 12.07 g/L, respectively. In the study of the influence of the galactose as inhibitor of the lactose hydrolysis, the competitive inhibition model was adjusted to the experimental results and the values of Vm, Km and Ki were 0,77 U, 12,07 g/L and 4,94 g/L respectively. |
publishDate |
2009 |
dc.date.issued.fl_str_mv |
2009-08-28 |
dc.date.available.fl_str_mv |
2010-03-02 2016-06-22T18:41:38Z |
dc.date.accessioned.fl_str_mv |
2016-06-22T18:41:38Z |
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 |
GUIDINI, Carla Zanella. Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica. 2009. 136 f. Dissertação (Mestrado em Engenharias) - Universidade Federal de Uberlândia, Uberlândia, 2009. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufu.br/handle/123456789/15134 |
identifier_str_mv |
GUIDINI, Carla Zanella. Imobilização de β-galactosidase de Aspergillus oryzae em resinas de troca iônica. 2009. 136 f. Dissertação (Mestrado em Engenharias) - Universidade Federal de Uberlândia, Uberlândia, 2009. |
url |
https://repositorio.ufu.br/handle/123456789/15134 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Engenharia Química |
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UFU |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Engenharias |
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Universidade Federal de Uberlândia |
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UFU |
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