Caracterização parcial e imobilização covalente da Neutrase® comercial
Autor(a) principal: | |
---|---|
Data de Publicação: | 2023 |
Tipo de documento: | Trabalho de conclusão de curso |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/17477 |
Resumo: | The use of enzymes as agents for modifying the functional properties of proteins has been widespread, mainly in the food industry. One of the most studied groups for this purpose are the proteases. Proteases are responsible for breaking peptide bonds between amino acids in proteins, producing smaller peptides and functional free amino acids. The use of the Neutrase® enzyme is explained by the fact that it is a protease that hydrolyses whey proteins, a highly polluting by-product and, most of the time, discarded in the dairy industries, aiming at obtaining derivatives with better functional properties. The general objective of this work was to partially characterize the crude extract of commercial Neutrase® derived from a Bacillus sp. The results obtained showed in the first enzymatic assay of free neutrase, its activity, determined through spectrophotometric analysis, was 27.9 U/mL. The total protein dosage found in the assay resulted in a value of 0.94 mg/mL. At the optimal activity temperature, 55 °C, the protease remained stable for a few hours. In addition, with the use of metallic ions, a characteristic of the existence of metalloproteases in the extract was observed. Regarding covalent immobilization, two supports were tested and the activity values of the Neutrase-Glyoxyl-Cob Corn (NGSM) and Neutrase Glyoxyl-Agarose (NGA) derivatives were 1.04 U/g and 1.15U/g, respectively. In addition to immobilization yields below expectations, unsatisfactory values for the methodology used. Despite this, future studies with other types of immobilization and functionalization of the support may favor the hydrolytic activity of the enzyme, a fact that still makes it a possible potential for new industrial applications. |
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Manzolli, Bruna Luiza de JesusMonti, Rubenshttp://lattes.cnpq.br/5962867835836749Novo-Mansur, Maria Teresa Marqueshttp://lattes.cnpq.br/1198926223396748http://lattes.cnpq.br/3652881556946374042fd67b-bff3-4842-bc63-7987bc90851a2023-03-13T11:58:58Z2023-03-13T11:58:58Z2023-02-02MANZOLLI, Bruna Luiza de Jesus. Caracterização parcial e imobilização covalente da Neutrase® comercial. 2023. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17477.https://repositorio.ufscar.br/handle/ufscar/17477The use of enzymes as agents for modifying the functional properties of proteins has been widespread, mainly in the food industry. One of the most studied groups for this purpose are the proteases. Proteases are responsible for breaking peptide bonds between amino acids in proteins, producing smaller peptides and functional free amino acids. The use of the Neutrase® enzyme is explained by the fact that it is a protease that hydrolyses whey proteins, a highly polluting by-product and, most of the time, discarded in the dairy industries, aiming at obtaining derivatives with better functional properties. The general objective of this work was to partially characterize the crude extract of commercial Neutrase® derived from a Bacillus sp. The results obtained showed in the first enzymatic assay of free neutrase, its activity, determined through spectrophotometric analysis, was 27.9 U/mL. The total protein dosage found in the assay resulted in a value of 0.94 mg/mL. At the optimal activity temperature, 55 °C, the protease remained stable for a few hours. In addition, with the use of metallic ions, a characteristic of the existence of metalloproteases in the extract was observed. Regarding covalent immobilization, two supports were tested and the activity values of the Neutrase-Glyoxyl-Cob Corn (NGSM) and Neutrase Glyoxyl-Agarose (NGA) derivatives were 1.04 U/g and 1.15U/g, respectively. In addition to immobilization yields below expectations, unsatisfactory values for the methodology used. Despite this, future studies with other types of immobilization and functionalization of the support may favor the hydrolytic activity of the enzyme, a fact that still makes it a possible potential for new industrial applications.A utilização de enzimas como agentes de modificação de propriedades funcionais de proteínas tem se difundido bastante, principalmente, nas indústrias de alimentos. Um dos grupos mais estudados para este fim são as proteases. Proteases são responsáveis pelas quebras de ligações peptídicas entre os aminoácidos das proteínas, produzindo peptídeos menores e aminoácidos livres funcionais. O uso da enzima Neutrase® se explica por ser uma protease que hidrolisa proteínas do soro de leite, um subproduto altamente poluente e, na maioria das vezes, descartado nas indústrias de laticínios, visando à obtenção de derivados com melhores propriedades funcionais. O objetivo geral deste trabalho consistiu em caracterizar parcialmente o extrato bruto da Neutrase® comercial derivada de um Bacillus sp. Os resultados obtidos mostraram no primeiro ensaio enzimático da neutrase livre, sua atividade, determinada através de análise espectrofométrica, foi de 27,9 U/mL. Já a dosagem de proteínas totais encontrada no ensaio resultou em um valor de 0,94 mg/mL. Na temperatura ótima de atividade, 55 °C, a protease manteve-se estável por algumas horas. Além disso, com o uso de íons metálicos observou-se uma característica da existência de metaloproteases no extrato. Em relação a imobilização covalente, foram testados dois suportes e os valores de atividade dos derivados Neutrase-Glioxil-Sabugo de milho (NGSM) e Neutrase-Glioxil-Agarose (NGA) foram 1,04 U/g e 1,15 U/g, respectivamente. Além de rendimentos de imobilização abaixo do esperado, valores não satisfatórios para a metodologia utilizada. Apesar disso, estudos futuros com outros tipos de imobilização e funcionalização do suporte podem favorecer a atividade hidrolítica da enzima, fato que ainda a torna um possível potencial para novas aplicações industriais.Não recebi financiamentoporUniversidade Federal de São CarlosCâmpus São CarlosBiotecnologia - BiotecUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessProteaseNeutraseImobilização covalenteHidróliseEnzimaEnzymeHydrolysisCovalent immobilizationCIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIACaracterização parcial e imobilização covalente da Neutrase® comercialPartial characterization and covalent immobilization of commercial Neutrase®info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesis6006005ded81db-48d2-4de7-b91a-256d87412f75reponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdf564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdfTrabalho de Conclusão de Cursoapplication/pdf1159154https://repositorio.ufscar.br/bitstream/ufscar/17477/1/564486_Bruna%20Luiza%20de%20Jesus%20Manzolli_TCCFINAL.pdf501880a23d9c1e39b8e6c7379aecda0eMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8810https://repositorio.ufscar.br/bitstream/ufscar/17477/2/license_rdff337d95da1fce0a22c77480e5e9a7aecMD52TEXT564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdf.txt564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdf.txtExtracted texttext/plain73089https://repositorio.ufscar.br/bitstream/ufscar/17477/3/564486_Bruna%20Luiza%20de%20Jesus%20Manzolli_TCCFINAL.pdf.txtc3552f4a1b77a0fe3399bee0977f0b97MD53THUMBNAIL564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdf.jpg564486_Bruna Luiza de Jesus Manzolli_TCCFINAL.pdf.jpgIM Thumbnailimage/jpeg5649https://repositorio.ufscar.br/bitstream/ufscar/17477/4/564486_Bruna%20Luiza%20de%20Jesus%20Manzolli_TCCFINAL.pdf.jpgb9d4ee810b476bbf5b48e60e17d6aeb2MD54ufscar/174772023-09-18 18:32:29.604oai:repositorio.ufscar.br:ufscar/17477Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:32:29Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
dc.title.alternative.eng.fl_str_mv |
Partial characterization and covalent immobilization of commercial Neutrase® |
title |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
spellingShingle |
Caracterização parcial e imobilização covalente da Neutrase® comercial Manzolli, Bruna Luiza de Jesus Protease Neutrase Imobilização covalente Hidrólise Enzima Enzyme Hydrolysis Covalent immobilization CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
title_short |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
title_full |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
title_fullStr |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
title_full_unstemmed |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
title_sort |
Caracterização parcial e imobilização covalente da Neutrase® comercial |
author |
Manzolli, Bruna Luiza de Jesus |
author_facet |
Manzolli, Bruna Luiza de Jesus |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/3652881556946374 |
dc.contributor.author.fl_str_mv |
Manzolli, Bruna Luiza de Jesus |
dc.contributor.advisor1.fl_str_mv |
Monti, Rubens |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5962867835836749 |
dc.contributor.advisor-co1.fl_str_mv |
Novo-Mansur, Maria Teresa Marques |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/1198926223396748 |
dc.contributor.authorID.fl_str_mv |
042fd67b-bff3-4842-bc63-7987bc90851a |
contributor_str_mv |
Monti, Rubens Novo-Mansur, Maria Teresa Marques |
dc.subject.por.fl_str_mv |
Protease Neutrase Imobilização covalente Hidrólise Enzima |
topic |
Protease Neutrase Imobilização covalente Hidrólise Enzima Enzyme Hydrolysis Covalent immobilization CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
dc.subject.eng.fl_str_mv |
Enzyme Hydrolysis Covalent immobilization |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
description |
The use of enzymes as agents for modifying the functional properties of proteins has been widespread, mainly in the food industry. One of the most studied groups for this purpose are the proteases. Proteases are responsible for breaking peptide bonds between amino acids in proteins, producing smaller peptides and functional free amino acids. The use of the Neutrase® enzyme is explained by the fact that it is a protease that hydrolyses whey proteins, a highly polluting by-product and, most of the time, discarded in the dairy industries, aiming at obtaining derivatives with better functional properties. The general objective of this work was to partially characterize the crude extract of commercial Neutrase® derived from a Bacillus sp. The results obtained showed in the first enzymatic assay of free neutrase, its activity, determined through spectrophotometric analysis, was 27.9 U/mL. The total protein dosage found in the assay resulted in a value of 0.94 mg/mL. At the optimal activity temperature, 55 °C, the protease remained stable for a few hours. In addition, with the use of metallic ions, a characteristic of the existence of metalloproteases in the extract was observed. Regarding covalent immobilization, two supports were tested and the activity values of the Neutrase-Glyoxyl-Cob Corn (NGSM) and Neutrase Glyoxyl-Agarose (NGA) derivatives were 1.04 U/g and 1.15U/g, respectively. In addition to immobilization yields below expectations, unsatisfactory values for the methodology used. Despite this, future studies with other types of immobilization and functionalization of the support may favor the hydrolytic activity of the enzyme, a fact that still makes it a possible potential for new industrial applications. |
publishDate |
2023 |
dc.date.accessioned.fl_str_mv |
2023-03-13T11:58:58Z |
dc.date.available.fl_str_mv |
2023-03-13T11:58:58Z |
dc.date.issued.fl_str_mv |
2023-02-02 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
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bachelorThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
MANZOLLI, Bruna Luiza de Jesus. Caracterização parcial e imobilização covalente da Neutrase® comercial. 2023. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17477. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/17477 |
identifier_str_mv |
MANZOLLI, Bruna Luiza de Jesus. Caracterização parcial e imobilização covalente da Neutrase® comercial. 2023. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2023. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17477. |
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https://repositorio.ufscar.br/handle/ufscar/17477 |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
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openAccess |
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Universidade Federal de São Carlos Câmpus São Carlos Biotecnologia - Biotec |
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Universidade Federal de São Carlos Câmpus São Carlos Biotecnologia - Biotec |
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