Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis

Detalhes bibliográficos
Autor(a) principal: Albuquerque, Tiago Lima de
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/30944
Resumo: In the last decades, due to the need to assure higher sustainability in industrial processes, the treatment of the waste deriving from the food industry has received unprecedented attention. An important sector in the business is dairy, which holds a prominent ranking due to the large volume of products processed. Whey, the main by-product of cheese, imposes considerable environmental risks, especially due to its the residual lactose, deeming pretreatment operations indispensable before disposal as an effluent. Among the promising alternatives for whey utilization, it can be highlighted its use as a raw material for the biotechnological production of products with high-added value, which includes the enzymatic hydrolysis of the lactose present for synthesizing oligosaccharides and other molecules with prebiotic function, such as lactulose. The latter has received considerable attention in scientific research due to its numerous uses in the food and pharmaceutical industries, for example, in the treatment of constipation and in the prevention and treatment of hepatic encephalopathy. Given the aforementioned scenario, the objective of the present study was to evaluate the employment of whey in the production of lactulose by an enzymatic route, using β-galactosidase from Kluyveromyces lactis. The results led to the determination of optimal conditions for the production of lactulose using a lactose/fructose-based synthetic medium, which initially were: lactose/fructose ratio of 15/30% (w/v); initial substrate concentration (lactose and fructose) of 200 g/L, temperature of 50 °C, enzymatic load of 7 U/mL, pH 7.0 and with the addition of 0.01% w/v Mg2+. The maximum concentration achieved was 22.59 ± 0.11 g/L of lactulose. Thereafter, the enzymatic lactulose production was investigated (with enzymes soluble and immobilized onto the previouslyselected support chitosan) using whey as raw material. The experiments presented promising results, given the high concentrations of lactulose obtained in the whey. It was concluded that the highest production was achieved when the whey had been treated for protein precipitation and elimination, yielding a total concentration of 17.32 ± 0.88 g/L of lactulose. The reuse cycles were also evaluated using the immobilized enzyme, with a high operational stability, with a synthesis of 63% of lactulose in the third cycle and about 25% at the end of the sixth cycle of reuse. From the hydrolysis of lactose, in addition to the liberated galactose, high amounts of glucose were generated in the reaction medium. As such, in an attempt to take advantage of these, the production of lactulose and ethanol was also evaluated in a single batch using enzymatic (by β -galactosidase from K. lactis) and fermentative (by Saccharomyces cerevisiae) routes, respectively. The results were potentially promising, since it was possible to produce lactulose and ethanol in the same batch at the maximum concentrations of 17.50 ± 0.76 g/L and 8.79 ± 0.65 g/L, respectively. Therefore, it can be concluded that it is possible to use the lactose present in the residual whey from the food industry for the production of lactulose, providing an appropriate destination for this waste, and consequently, reducing the environmental impact it can exert. Furthermore, this practice would result in the production of a substance with higher added value due to its extensive use in the food and pharmaceutical industries.
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spelling Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactisEngenharia químicaLeite - ReaproveitamentoLactuloseEnzimasResíduos industriaisAlimentos - Indústria - SubprodutosImmobilizationReutilization of wheyIn the last decades, due to the need to assure higher sustainability in industrial processes, the treatment of the waste deriving from the food industry has received unprecedented attention. An important sector in the business is dairy, which holds a prominent ranking due to the large volume of products processed. Whey, the main by-product of cheese, imposes considerable environmental risks, especially due to its the residual lactose, deeming pretreatment operations indispensable before disposal as an effluent. Among the promising alternatives for whey utilization, it can be highlighted its use as a raw material for the biotechnological production of products with high-added value, which includes the enzymatic hydrolysis of the lactose present for synthesizing oligosaccharides and other molecules with prebiotic function, such as lactulose. The latter has received considerable attention in scientific research due to its numerous uses in the food and pharmaceutical industries, for example, in the treatment of constipation and in the prevention and treatment of hepatic encephalopathy. Given the aforementioned scenario, the objective of the present study was to evaluate the employment of whey in the production of lactulose by an enzymatic route, using β-galactosidase from Kluyveromyces lactis. The results led to the determination of optimal conditions for the production of lactulose using a lactose/fructose-based synthetic medium, which initially were: lactose/fructose ratio of 15/30% (w/v); initial substrate concentration (lactose and fructose) of 200 g/L, temperature of 50 °C, enzymatic load of 7 U/mL, pH 7.0 and with the addition of 0.01% w/v Mg2+. The maximum concentration achieved was 22.59 ± 0.11 g/L of lactulose. Thereafter, the enzymatic lactulose production was investigated (with enzymes soluble and immobilized onto the previouslyselected support chitosan) using whey as raw material. The experiments presented promising results, given the high concentrations of lactulose obtained in the whey. It was concluded that the highest production was achieved when the whey had been treated for protein precipitation and elimination, yielding a total concentration of 17.32 ± 0.88 g/L of lactulose. The reuse cycles were also evaluated using the immobilized enzyme, with a high operational stability, with a synthesis of 63% of lactulose in the third cycle and about 25% at the end of the sixth cycle of reuse. From the hydrolysis of lactose, in addition to the liberated galactose, high amounts of glucose were generated in the reaction medium. As such, in an attempt to take advantage of these, the production of lactulose and ethanol was also evaluated in a single batch using enzymatic (by β -galactosidase from K. lactis) and fermentative (by Saccharomyces cerevisiae) routes, respectively. The results were potentially promising, since it was possible to produce lactulose and ethanol in the same batch at the maximum concentrations of 17.50 ± 0.76 g/L and 8.79 ± 0.65 g/L, respectively. Therefore, it can be concluded that it is possible to use the lactose present in the residual whey from the food industry for the production of lactulose, providing an appropriate destination for this waste, and consequently, reducing the environmental impact it can exert. Furthermore, this practice would result in the production of a substance with higher added value due to its extensive use in the food and pharmaceutical industries.Nas últimas décadas, devido a necessidade de alcançar maior sustentabilidade em processos industriais, mais atenção vem sendo conferida ao tratamento de resíduos provenientes da Indústria de Alimentos. Um importante setor dessa indústria é o de laticínios, que ocupa lugar de destaque devido ao grande volume de produtos processados. O soro de leite, principal subproduto da fabricação de queijos, possui nocividade ambiental considerável, especialmente pela lactose residual presente, tornando indispensável seu pré-tratamento antes de ser descartado como efluente. Dentre as alternativas promissoras para a utilização desse soro, destaca-se seu emprego como matéria-prima para a produção biotecnológica de produtos com maior valor agregado, o que poderia incluir a hidrólise enzimática da lactose presente para síntese de oligossacarídeos e substâncias com função prebiótica, como a lactulose. Essa última substância tem recebido muita atenção nas pesquisas científicas devido aos seus diversos usos na indústria de alimentos e farmacêutica, atuando, por exemplo, no tratamento de constipação e na prevenção e o tratamento de encefalopatia hepática. O objetivo do presente estudo, de acordo com as alegações citadas anteriormente, foi avaliar o aproveitamento de soro de leite para a produção de lactulose por via enzimática, utilizando a enzima β-galactosidase de Kluyveromyces lactis. Os resultados levaram a determinação das condições ótimas para a produção de lactulose utilizando, inicialmente, meio sintético a base de lactose e frutose, que foram: razão lactose/frutose de 15/30 % (m/v); concentração inicial dos substratos (lactose e frutose) de 200 g/L, temperatura de 50 °C, carga enzimática de 7 U/mL, pH 7 e adição de 0,01% m/v de Mg2+. A concentração máxima alcançada foi de 22,59 ± 0,11 g/L de lactulose. A partir de então, investigou-se a produção de lactulose por via enzimática (tanto com a enzima solúvel como imobilizada no suporte previamente selecionado – quitosana) utilizando-se o soro de leite como matéria-pima. Os experimentos apresentaram resultados promissores, uma vez que elevadas concentrações de lactulose foram obtidas a partir do soro. Concluiu-se que a maior produção foi alcançada no soro de leite tratado para precipitação e eliminação de proteína, atingindo-se uma concentração total de 17,32 ± 0,88 g/L de lactulose. Também se avaliaram os ciclos de reuso utilizando a enzima imobilizada, observando-se elevada estabilidade operacional, com síntese, em relação ao ciclo inicial, de 63% de lactulose no terceiro, e cerca 25% ao final do sexto ciclo de reuso. A partir da hidrólise da lactose, além da galactose liberada, geraram-se altas quantidades de glicose no meio reacional, assim, no intento de aproveitar essa glicose, avaliou-se a produção em uma única batelada de lactulose e etanol via enzimática (por β-galactosidase de K. lactis) e fermentativa (por Saccharomyces cerevisiae), respectivamente. Os resultados encontrados foram potencialmente promissores, visto que foi possível produzir lactulose e etanol em uma mesma batelada nas concentrações máximas de 17,50 ± 0,76 g/L e 8,79 ± 0,65 g/L, respectivamente. Conclui-se, portanto, que é possível a utilização da lactose presente no soro de leite residual da Indústria de Alimentos para a produção de lactulose, o que proporcionaria um destino adequado a esse resíduo, com a redução dos danos ambientais e de custos que pode causar. Além da produção de uma substância com maior valor agregado devido a seu extenso uso em indústrias alimentícias e farmacêuticas.Rocha, Maria Valderez PonteGonçalves, Luciana Rocha BarrosAlbuquerque, Tiago Lima de2018-04-11T17:16:49Z2018-04-11T17:16:49Z2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfALBUQUERQUE, T. L. Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis. 2018. 155 f. Tese (Doutorado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.http://www.repositorio.ufc.br/handle/riufc/30944porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-09-23T13:34:55Zoai:repositorio.ufc.br:riufc/30944Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:27:49.104277Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
title Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
spellingShingle Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
Albuquerque, Tiago Lima de
Engenharia química
Leite - Reaproveitamento
Lactulose
Enzimas
Resíduos industriais
Alimentos - Indústria - Subprodutos
Immobilization
Reutilization of whey
title_short Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
title_full Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
title_fullStr Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
title_full_unstemmed Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
title_sort Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis
author Albuquerque, Tiago Lima de
author_facet Albuquerque, Tiago Lima de
author_role author
dc.contributor.none.fl_str_mv Rocha, Maria Valderez Ponte
Gonçalves, Luciana Rocha Barros
dc.contributor.author.fl_str_mv Albuquerque, Tiago Lima de
dc.subject.por.fl_str_mv Engenharia química
Leite - Reaproveitamento
Lactulose
Enzimas
Resíduos industriais
Alimentos - Indústria - Subprodutos
Immobilization
Reutilization of whey
topic Engenharia química
Leite - Reaproveitamento
Lactulose
Enzimas
Resíduos industriais
Alimentos - Indústria - Subprodutos
Immobilization
Reutilization of whey
description In the last decades, due to the need to assure higher sustainability in industrial processes, the treatment of the waste deriving from the food industry has received unprecedented attention. An important sector in the business is dairy, which holds a prominent ranking due to the large volume of products processed. Whey, the main by-product of cheese, imposes considerable environmental risks, especially due to its the residual lactose, deeming pretreatment operations indispensable before disposal as an effluent. Among the promising alternatives for whey utilization, it can be highlighted its use as a raw material for the biotechnological production of products with high-added value, which includes the enzymatic hydrolysis of the lactose present for synthesizing oligosaccharides and other molecules with prebiotic function, such as lactulose. The latter has received considerable attention in scientific research due to its numerous uses in the food and pharmaceutical industries, for example, in the treatment of constipation and in the prevention and treatment of hepatic encephalopathy. Given the aforementioned scenario, the objective of the present study was to evaluate the employment of whey in the production of lactulose by an enzymatic route, using β-galactosidase from Kluyveromyces lactis. The results led to the determination of optimal conditions for the production of lactulose using a lactose/fructose-based synthetic medium, which initially were: lactose/fructose ratio of 15/30% (w/v); initial substrate concentration (lactose and fructose) of 200 g/L, temperature of 50 °C, enzymatic load of 7 U/mL, pH 7.0 and with the addition of 0.01% w/v Mg2+. The maximum concentration achieved was 22.59 ± 0.11 g/L of lactulose. Thereafter, the enzymatic lactulose production was investigated (with enzymes soluble and immobilized onto the previouslyselected support chitosan) using whey as raw material. The experiments presented promising results, given the high concentrations of lactulose obtained in the whey. It was concluded that the highest production was achieved when the whey had been treated for protein precipitation and elimination, yielding a total concentration of 17.32 ± 0.88 g/L of lactulose. The reuse cycles were also evaluated using the immobilized enzyme, with a high operational stability, with a synthesis of 63% of lactulose in the third cycle and about 25% at the end of the sixth cycle of reuse. From the hydrolysis of lactose, in addition to the liberated galactose, high amounts of glucose were generated in the reaction medium. As such, in an attempt to take advantage of these, the production of lactulose and ethanol was also evaluated in a single batch using enzymatic (by β -galactosidase from K. lactis) and fermentative (by Saccharomyces cerevisiae) routes, respectively. The results were potentially promising, since it was possible to produce lactulose and ethanol in the same batch at the maximum concentrations of 17.50 ± 0.76 g/L and 8.79 ± 0.65 g/L, respectively. Therefore, it can be concluded that it is possible to use the lactose present in the residual whey from the food industry for the production of lactulose, providing an appropriate destination for this waste, and consequently, reducing the environmental impact it can exert. Furthermore, this practice would result in the production of a substance with higher added value due to its extensive use in the food and pharmaceutical industries.
publishDate 2018
dc.date.none.fl_str_mv 2018-04-11T17:16:49Z
2018-04-11T17:16:49Z
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 ALBUQUERQUE, T. L. Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis. 2018. 155 f. Tese (Doutorado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.
http://www.repositorio.ufc.br/handle/riufc/30944
identifier_str_mv ALBUQUERQUE, T. L. Produção enzimática de lactulose a partir de soro de leite utilizando a enzima β-galactosidase de Kluyveromyces lactis. 2018. 155 f. Tese (Doutorado em Engenharia Química)-Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2018.
url http://www.repositorio.ufc.br/handle/riufc/30944
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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)
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reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
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repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
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