Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis

Detalhes bibliográficos
Autor(a) principal: Schultz, Guilhermina
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/14203
Resumo: Galacto-oligosaccharides (GOS) have a high commercial value and are used in the food industry. GOS are non-digestible oligosaccharides (NDO), which are considered prebiotics. They can be produced enzymatically from substrates rich in lactose (whey, whey permeate or lactose) using the enzyme β-galactosidase as a catalyst, through a kinetically controlled reaction characterized by competition between GOS synthesis reactions (transgalactosylation) and lactose hydrolysis. In these substrates there are considerable concentrations of cations (Na+, Mg+2, Ca+2 and K+) that can affect the activity of β-galactosidase. Thus, the objective of this work was to study the kinetics of the enzymatic synthesis of GOS using lactose and whey permeate as substrate, using the free enzyme β-galactosidase from Kluyveromyces lactis (Lactozym 3000 L HP G) as a catalyst, which has GRAS status, to arrive at mathematical models that allow the use of computational tools in order to optimize the process. To achieve this goal, several reactions were conducted. In the initial rate assays, most of the experiments were carried out on 220 g/L of lactose (pure, or present in whey permeate) and chloride salts were added when using pure lactose, to verify their influence on the initial rate of synthesis and hydrolysis, total activity and selectivity. The results obtained indicate that, within the studied region, sodium, among all the ions tested, was the one that caused the greatest selectivity. Calcium inhibited the initial rate of enzyme synthesis and hydrolysis. In media containing potassium together with sodium, the effect of sodium inhibition on the initial rate of hydrolysis was even more pronounced. A similar effect occurred when potassium and calcium were used, since the inhibition caused by calcium, in the total activity, initial rate of synthesis and hydrolysis seems to be potentiated by the presence of potassium. For the kinetic model, fit were made by non-linear regression, in which several initial concentrations of lactose and enzyme were used. The model has nine adjustable parameters and characterizes the formation of Glb, tri, tetrasaccharides and enzymatic inactivation. A bootstrap method, based on the residues obtained during the adjustment procedure, is employed to calculate the confidence intervals for the parameters. This model fits well with the experimental data. As a result, this model can be useful for optimization of bioreactors and process design, as well as process control.
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spelling Schultz, GuilherminaRibeiro, Marcelo Perencin de Arrudahttp://lattes.cnpq.br/0381402687491195Giordano, Roberto de Camposhttp://lattes.cnpq.br/0834668419587001http://lattes.cnpq.br/6565060554031053b1f1bbcd-c222-465b-a65d-fa837f340cc02021-04-30T11:27:48Z2021-04-30T11:27:48Z2020-12-22SCHULTZ, Guilhermina. Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/14203.https://repositorio.ufscar.br/handle/ufscar/14203Galacto-oligosaccharides (GOS) have a high commercial value and are used in the food industry. GOS are non-digestible oligosaccharides (NDO), which are considered prebiotics. They can be produced enzymatically from substrates rich in lactose (whey, whey permeate or lactose) using the enzyme β-galactosidase as a catalyst, through a kinetically controlled reaction characterized by competition between GOS synthesis reactions (transgalactosylation) and lactose hydrolysis. In these substrates there are considerable concentrations of cations (Na+, Mg+2, Ca+2 and K+) that can affect the activity of β-galactosidase. Thus, the objective of this work was to study the kinetics of the enzymatic synthesis of GOS using lactose and whey permeate as substrate, using the free enzyme β-galactosidase from Kluyveromyces lactis (Lactozym 3000 L HP G) as a catalyst, which has GRAS status, to arrive at mathematical models that allow the use of computational tools in order to optimize the process. To achieve this goal, several reactions were conducted. In the initial rate assays, most of the experiments were carried out on 220 g/L of lactose (pure, or present in whey permeate) and chloride salts were added when using pure lactose, to verify their influence on the initial rate of synthesis and hydrolysis, total activity and selectivity. The results obtained indicate that, within the studied region, sodium, among all the ions tested, was the one that caused the greatest selectivity. Calcium inhibited the initial rate of enzyme synthesis and hydrolysis. In media containing potassium together with sodium, the effect of sodium inhibition on the initial rate of hydrolysis was even more pronounced. A similar effect occurred when potassium and calcium were used, since the inhibition caused by calcium, in the total activity, initial rate of synthesis and hydrolysis seems to be potentiated by the presence of potassium. For the kinetic model, fit were made by non-linear regression, in which several initial concentrations of lactose and enzyme were used. The model has nine adjustable parameters and characterizes the formation of Glb, tri, tetrasaccharides and enzymatic inactivation. A bootstrap method, based on the residues obtained during the adjustment procedure, is employed to calculate the confidence intervals for the parameters. This model fits well with the experimental data. As a result, this model can be useful for optimization of bioreactors and process design, as well as process control.Galacto-oligossacarídeos (GOS) apresentam um alto valor comercial e são usados na indústria de alimentos. GOS são oligossacarídeos não digeríveis (NDO), que são considerados prebióticos. Podem ser produzidos enzimaticamente a partir de substratos ricos em lactose (soro de leite, permeado do soro de leite ou lactose) usando a enzima β-galactosidase como catalisador, por meio de uma reação controlada cineticamente caracterizada pela competição entre as reações de síntese de GOS (transgalactosilação) e hidrólise da lactose. Nesses substratos há concentrações consideráveis de cátions (Na+, Mg+2, Ca+2 e K+) que podem afetar a atividade da β-galactosidase. Dessa forma, o objetivo desse trabalho foi estudar a cinética da síntese enzimática de GOS utilizando como substrato lactose e permeado do soro de leite empregando como catalisador a enzima β-galactosidase livre de Kluyveromyces lactis (Lactozym 3000 L HP G) que possui status GRAS para se chegar a modelos matemáticos que viabilizem empregar ferramentas computacionais com intenção de otimização do processo. Para atingir esse objetivo foram conduzidas diversas reações. Nos ensaios de velocidades iniciais, a maior parte dos experimentos foram realizados em 220 g/L de lactose (pura, ou presente em permeado do soro de leite) e sais de cloreto foram adicionados quando utilizada lactose pura, para verificar a influência deles sobre a velocidade inicial de síntese e hidrólise, atividade total e seletividade. Os resultados obtidos indicam que, dentro da região estudada, o sódio, dentre todos os íons testados, foi o que provocou maior seletividade. O cálcio inibiu a velocidade inicial de síntese e hidrólise da enzima. Em meios contendo potássio juntamente com sódio, o efeito de inibição pelo sódio na velocidade inicial de hidrólise foi ainda mais acentuado. Efeito parecido ocorreu, quando foi utilizado potássio e cálcio, pois a inibição causada pelo cálcio, na atividade total, velocidade inicial de síntese e hidrólise parece ser potencializado pela presença do potássio. Para o modelo cinético, os ajustes foram feitos por regressão não linear, no qual foram utilizadas diversas concentrações inciais de lactose e enzima. O modelo apresenta nove parâmetros ajustáveis e caracteriza a formação de Glb, tri, tetrassacarídeos e a inativação enzimática. Um método de bootstrap, com base nos resíduos obtidos durante o procedimento de ajuste, é empregado para calcular os intervalos de confiança dos parâmetros. Este modelo se ajusta bem aos dados experimentais. Como resultado, este modelo pode ser útil para otimização de biorreatores e projeto de processos, bem como controle de processos.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)código financeiro 001FAPESP: 2018/04933-5porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Engenharia Química - PPGEQUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessGalactobioseGalacto-oligossacarídeosβ-galactosidaseKluyveromyces lactisModelagemGalacto-oligosaccharidesModelingENGENHARIAS::ENGENHARIA QUIMICAModelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactisKinetic modeling of synthesis galacto-oligosaccharides using β-galactosidase from Kluyveromyces lactisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisf459889e-82d5-45b8-9703-0d4926219d13reponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALSCHULTZ, 2020 - versão final.pdfSCHULTZ, 2020 - versão final.pdfapplication/pdf2378996https://repositorio.ufscar.br/bitstream/ufscar/14203/5/SCHULTZ%2c%202020%20-%20vers%c3%a3o%20final.pdf14f66c65550949adc6d36c07bb53e3b5MD55carta versão final-Guilhermina.pdfcarta versão final-Guilhermina.pdfapplication/pdf145747https://repositorio.ufscar.br/bitstream/ufscar/14203/6/carta%20vers%c3%a3o%20final-Guilhermina.pdf2b65aba5a18c2088d061bd7c569db26aMD56CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufscar.br/bitstream/ufscar/14203/7/license_rdfe39d27027a6cc9cb039ad269a5db8e34MD57TEXTSCHULTZ, 2020 - versão final.pdf.txtSCHULTZ, 2020 - versão final.pdf.txtExtracted texttext/plain232021https://repositorio.ufscar.br/bitstream/ufscar/14203/8/SCHULTZ%2c%202020%20-%20vers%c3%a3o%20final.pdf.txtb4258000cec492e4bc8d51fc1077a009MD58carta versão final-Guilhermina.pdf.txtcarta versão final-Guilhermina.pdf.txtExtracted texttext/plain1486https://repositorio.ufscar.br/bitstream/ufscar/14203/10/carta%20vers%c3%a3o%20final-Guilhermina.pdf.txt247846f6a261ca637074874e6d68b919MD510THUMBNAILSCHULTZ, 2020 - versão final.pdf.jpgSCHULTZ, 2020 - versão final.pdf.jpgIM Thumbnailimage/jpeg5171https://repositorio.ufscar.br/bitstream/ufscar/14203/9/SCHULTZ%2c%202020%20-%20vers%c3%a3o%20final.pdf.jpg5464a5fdcd583f038e3f9e22d51c5eb8MD59carta versão final-Guilhermina.pdf.jpgcarta versão final-Guilhermina.pdf.jpgIM Thumbnailimage/jpeg6545https://repositorio.ufscar.br/bitstream/ufscar/14203/11/carta%20vers%c3%a3o%20final-Guilhermina.pdf.jpg48f61700dfcfa706774a6a40d2036e36MD511ufscar/142032023-09-18 18:32:09.77oai:repositorio.ufscar.br:ufscar/14203Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:32:09Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
dc.title.alternative.eng.fl_str_mv Kinetic modeling of synthesis galacto-oligosaccharides using β-galactosidase from Kluyveromyces lactis
title Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
spellingShingle Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
Schultz, Guilhermina
Galactobiose
Galacto-oligossacarídeos
β-galactosidase
Kluyveromyces lactis
Modelagem
Galacto-oligosaccharides
Modeling
ENGENHARIAS::ENGENHARIA QUIMICA
title_short Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
title_full Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
title_fullStr Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
title_full_unstemmed Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
title_sort Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis
author Schultz, Guilhermina
author_facet Schultz, Guilhermina
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/6565060554031053
dc.contributor.author.fl_str_mv Schultz, Guilhermina
dc.contributor.advisor1.fl_str_mv Ribeiro, Marcelo Perencin de Arruda
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/0381402687491195
dc.contributor.advisor-co1.fl_str_mv Giordano, Roberto de Campos
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/0834668419587001
dc.contributor.authorID.fl_str_mv b1f1bbcd-c222-465b-a65d-fa837f340cc0
contributor_str_mv Ribeiro, Marcelo Perencin de Arruda
Giordano, Roberto de Campos
dc.subject.por.fl_str_mv Galactobiose
Galacto-oligossacarídeos
β-galactosidase
Kluyveromyces lactis
Modelagem
topic Galactobiose
Galacto-oligossacarídeos
β-galactosidase
Kluyveromyces lactis
Modelagem
Galacto-oligosaccharides
Modeling
ENGENHARIAS::ENGENHARIA QUIMICA
dc.subject.eng.fl_str_mv Galacto-oligosaccharides
Modeling
dc.subject.cnpq.fl_str_mv ENGENHARIAS::ENGENHARIA QUIMICA
description Galacto-oligosaccharides (GOS) have a high commercial value and are used in the food industry. GOS are non-digestible oligosaccharides (NDO), which are considered prebiotics. They can be produced enzymatically from substrates rich in lactose (whey, whey permeate or lactose) using the enzyme β-galactosidase as a catalyst, through a kinetically controlled reaction characterized by competition between GOS synthesis reactions (transgalactosylation) and lactose hydrolysis. In these substrates there are considerable concentrations of cations (Na+, Mg+2, Ca+2 and K+) that can affect the activity of β-galactosidase. Thus, the objective of this work was to study the kinetics of the enzymatic synthesis of GOS using lactose and whey permeate as substrate, using the free enzyme β-galactosidase from Kluyveromyces lactis (Lactozym 3000 L HP G) as a catalyst, which has GRAS status, to arrive at mathematical models that allow the use of computational tools in order to optimize the process. To achieve this goal, several reactions were conducted. In the initial rate assays, most of the experiments were carried out on 220 g/L of lactose (pure, or present in whey permeate) and chloride salts were added when using pure lactose, to verify their influence on the initial rate of synthesis and hydrolysis, total activity and selectivity. The results obtained indicate that, within the studied region, sodium, among all the ions tested, was the one that caused the greatest selectivity. Calcium inhibited the initial rate of enzyme synthesis and hydrolysis. In media containing potassium together with sodium, the effect of sodium inhibition on the initial rate of hydrolysis was even more pronounced. A similar effect occurred when potassium and calcium were used, since the inhibition caused by calcium, in the total activity, initial rate of synthesis and hydrolysis seems to be potentiated by the presence of potassium. For the kinetic model, fit were made by non-linear regression, in which several initial concentrations of lactose and enzyme were used. The model has nine adjustable parameters and characterizes the formation of Glb, tri, tetrasaccharides and enzymatic inactivation. A bootstrap method, based on the residues obtained during the adjustment procedure, is employed to calculate the confidence intervals for the parameters. This model fits well with the experimental data. As a result, this model can be useful for optimization of bioreactors and process design, as well as process control.
publishDate 2020
dc.date.issued.fl_str_mv 2020-12-22
dc.date.accessioned.fl_str_mv 2021-04-30T11:27:48Z
dc.date.available.fl_str_mv 2021-04-30T11:27:48Z
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dc.identifier.citation.fl_str_mv SCHULTZ, Guilhermina. Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/14203.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/14203
identifier_str_mv SCHULTZ, Guilhermina. Modelagem cinética da síntese de galacto-oligossacarídeos usando a β-galactosidase de Kluyveromyces lactis. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/14203.
url https://repositorio.ufscar.br/handle/ufscar/14203
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http://creativecommons.org/licenses/by-nc-nd/3.0/br/
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dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Engenharia Química - PPGEQ
dc.publisher.initials.fl_str_mv UFSCar
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