Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso

Detalhes bibliográficos
Autor(a) principal: Pinheiro, Maísa Pessoa
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/31829
Resumo: The aim of this work was to immobilize Lecitase Ultra ® (LU) onto Immobead-350 (IB-350) support through different immobilization protocols, using unmodified support and modified support (activated with functional groups by several strategies) and also through immobilization by cross-linking. The different immobilization strategies in this work were carried out in order to modulate enzyme properties and to evaluate the effects of three parameters in the final enzyme conformation and biocatalyst properties: incubation time; different pH values in the ionization of functional groups of the enzyme and of the support; and blocking stage. In addition, biocatalysts formed by immobilization of LU onto supports activated with aldehyde, divinylsulfone or glutaraldehyde groups were evaluated for their catalytic versatility versus two different substrates. The results showed that all biocatalysts were active versus the hydrolysis of p-nitrophenyl butyrate (p-NPB), but Leci-GLU showed the highest derivative activity (17.39 U / g) and was the most stable preparation both at 50 °C (1.9 h) and in the presence of 30% acetonitrile (42.6 h). The derivatives were also tested for operational and storage stability. In this first assay, the biocatalysts maintained 100 % of their initial activity during 10 consecutive reaction cycles, while in storage stability all the derivatives remained with 80 % of the initial activity in 30 days of storage at 4 °C, only Leci-DVS reduced about 50 % of its initial activity. In order to test catalytic versatility, biocatalysts were applied in hydrolysis (methyl mandelate substrate) and transesterification (acetylation of benzyl alcohol) reactions. The product obtained from this last reaction is known as jasmine aroma and has high commercial value, in addition, it can be used in detergent and perfume industry. In relation to methyl mandelate hydrolysis, the biocatalysts were tested at different reaction conditions (pHs 5, 7 and 9) and the highest activity was obtained with the Leci-crossA derivative at pH 7 (28.4 U / mg). In benzyl alcohol acetylation the biocatalyst with the highest activity was Leci-GLU (2.1 mg).
id UFC-7_47ee2b084635ff266503214a31ce510b
oai_identifier_str oai:repositorio.ufc.br:riufc/31829
network_acronym_str UFC-7
network_name_str Repositório Institucional da Universidade Federal do Ceará (UFC)
repository_id_str
spelling Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporosoEngenharia químicaFosfolipases ABiocatalisadoresEnzimasLecitase UltraStability enzymeBiocatalystsThe aim of this work was to immobilize Lecitase Ultra ® (LU) onto Immobead-350 (IB-350) support through different immobilization protocols, using unmodified support and modified support (activated with functional groups by several strategies) and also through immobilization by cross-linking. The different immobilization strategies in this work were carried out in order to modulate enzyme properties and to evaluate the effects of three parameters in the final enzyme conformation and biocatalyst properties: incubation time; different pH values in the ionization of functional groups of the enzyme and of the support; and blocking stage. In addition, biocatalysts formed by immobilization of LU onto supports activated with aldehyde, divinylsulfone or glutaraldehyde groups were evaluated for their catalytic versatility versus two different substrates. The results showed that all biocatalysts were active versus the hydrolysis of p-nitrophenyl butyrate (p-NPB), but Leci-GLU showed the highest derivative activity (17.39 U / g) and was the most stable preparation both at 50 °C (1.9 h) and in the presence of 30% acetonitrile (42.6 h). The derivatives were also tested for operational and storage stability. In this first assay, the biocatalysts maintained 100 % of their initial activity during 10 consecutive reaction cycles, while in storage stability all the derivatives remained with 80 % of the initial activity in 30 days of storage at 4 °C, only Leci-DVS reduced about 50 % of its initial activity. In order to test catalytic versatility, biocatalysts were applied in hydrolysis (methyl mandelate substrate) and transesterification (acetylation of benzyl alcohol) reactions. The product obtained from this last reaction is known as jasmine aroma and has high commercial value, in addition, it can be used in detergent and perfume industry. In relation to methyl mandelate hydrolysis, the biocatalysts were tested at different reaction conditions (pHs 5, 7 and 9) and the highest activity was obtained with the Leci-crossA derivative at pH 7 (28.4 U / mg). In benzyl alcohol acetylation the biocatalyst with the highest activity was Leci-GLU (2.1 mg).A proposta desse trabalho foi imobilizar a Lecitase Ultra ® (LU) no suporte Immobead-350 (IB- 350) através de diferentes protocolos de imobilização, utilizando o suporte sem modificação e o suporte modificado (ativado com grupos funcionais por diversas estratégias) e, também, através da imobilização por cross-linking. Os diferentes protocolos foram executados com o intuito de modular as propriedades da enzima e avaliar o efeito que o tempo de incubação, a utilização de diferentes pHs na ionização dos grupos funcionais da enzima e do suporte, assim como a etapa de bloqueio, podem exercer na conformação final da enzima e nas propriedades dos biocatalisadores. Além disso, avaliou-se a versatilidade dos biocatalisadores imobilizados em suportes ativados com grupos epóxi, aldeído, divinilsulfona e glutaraldeído. Os resultados obtidos mostraram que todos os biocatalisadores produzidos foram ativos frente à hidrólise do p-nitrofenolbutirato (p-NPB), porém Leci-GLU apresentou a maior atividade do derivado (17,39 U/g) e foi a preparação mais estável na temperatura de 50 °C (1,9 h) e na presença de acetonitrila-30 % (42,6 h). Os derivados também foram testados quanto à estabilidade operacional e de estocagem. Nesse primeiro ensaio, os biocatalisadores mantiveram 100 % da sua atividade inicial durante 10 ciclos reacionais consecutivos, enquanto na estabilidade de estocagem, todos os derivados permaneceram com 80 % da atividade inicial em 30 dias de armazenamento a 4°C, exceto Leci-DVS que reduziu cerca de 50 % da sua atividade inicial. Para testar a versatilidade dos biocatalisadores, eles foram aplicados em reações de hidrólise (substrato mandelato de metila) e transesterificação (acetilação do álcool benzílico). O produto obtido dessa última reação é conhecido como aroma de jasmim e possui alto valor comercial, podendo ser utilizado na indústria de detergentes e perfumaria. Para esses ensaios, a hidrólise do mandelato de metila foi testada em diferentes pHs (5, 7 e 9) e a maior atividade foi obtida com o derivado Leci-crossA a pH 7 (28,4 U/mg), enquanto na acetilação do álcool benzílico o biocatalisador com maior atividade foi Leci-GLU (2,1 mg).Santos, José Cleiton Sousa dosGonçalves, Luciana Rocha BarrosPinheiro, Maísa Pessoa2018-05-10T12:44:56Z2018-05-10T12:44:56Z2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfPINHEIRO, M. P. Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso. 2018. 93 f. Dissertação (Mestrado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.http://www.repositorio.ufc.br/handle/riufc/31829porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-09-15T18:55:23Zoai:repositorio.ufc.br:riufc/31829Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:41:52.614097Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
title Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
spellingShingle Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
Pinheiro, Maísa Pessoa
Engenharia química
Fosfolipases A
Biocatalisadores
Enzimas
Lecitase Ultra
Stability enzyme
Biocatalysts
title_short Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
title_full Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
title_fullStr Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
title_full_unstemmed Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
title_sort Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso
author Pinheiro, Maísa Pessoa
author_facet Pinheiro, Maísa Pessoa
author_role author
dc.contributor.none.fl_str_mv Santos, José Cleiton Sousa dos
Gonçalves, Luciana Rocha Barros
dc.contributor.author.fl_str_mv Pinheiro, Maísa Pessoa
dc.subject.por.fl_str_mv Engenharia química
Fosfolipases A
Biocatalisadores
Enzimas
Lecitase Ultra
Stability enzyme
Biocatalysts
topic Engenharia química
Fosfolipases A
Biocatalisadores
Enzimas
Lecitase Ultra
Stability enzyme
Biocatalysts
description The aim of this work was to immobilize Lecitase Ultra ® (LU) onto Immobead-350 (IB-350) support through different immobilization protocols, using unmodified support and modified support (activated with functional groups by several strategies) and also through immobilization by cross-linking. The different immobilization strategies in this work were carried out in order to modulate enzyme properties and to evaluate the effects of three parameters in the final enzyme conformation and biocatalyst properties: incubation time; different pH values in the ionization of functional groups of the enzyme and of the support; and blocking stage. In addition, biocatalysts formed by immobilization of LU onto supports activated with aldehyde, divinylsulfone or glutaraldehyde groups were evaluated for their catalytic versatility versus two different substrates. The results showed that all biocatalysts were active versus the hydrolysis of p-nitrophenyl butyrate (p-NPB), but Leci-GLU showed the highest derivative activity (17.39 U / g) and was the most stable preparation both at 50 °C (1.9 h) and in the presence of 30% acetonitrile (42.6 h). The derivatives were also tested for operational and storage stability. In this first assay, the biocatalysts maintained 100 % of their initial activity during 10 consecutive reaction cycles, while in storage stability all the derivatives remained with 80 % of the initial activity in 30 days of storage at 4 °C, only Leci-DVS reduced about 50 % of its initial activity. In order to test catalytic versatility, biocatalysts were applied in hydrolysis (methyl mandelate substrate) and transesterification (acetylation of benzyl alcohol) reactions. The product obtained from this last reaction is known as jasmine aroma and has high commercial value, in addition, it can be used in detergent and perfume industry. In relation to methyl mandelate hydrolysis, the biocatalysts were tested at different reaction conditions (pHs 5, 7 and 9) and the highest activity was obtained with the Leci-crossA derivative at pH 7 (28.4 U / mg). In benzyl alcohol acetylation the biocatalyst with the highest activity was Leci-GLU (2.1 mg).
publishDate 2018
dc.date.none.fl_str_mv 2018-05-10T12:44:56Z
2018-05-10T12:44:56Z
2018
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 PINHEIRO, M. P. Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso. 2018. 93 f. Dissertação (Mestrado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.
http://www.repositorio.ufc.br/handle/riufc/31829
identifier_str_mv PINHEIRO, M. P. Desenvolvimento de biocatalisadores enzimáticos pela estabilização de Lecitase Ultra em suporte macroporoso. 2018. 93 f. Dissertação (Mestrado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.
url http://www.repositorio.ufc.br/handle/riufc/31829
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
_version_ 1813028910494908416