Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju

Detalhes bibliográficos
Ano de defesa: 2016
Autor(a) principal: Serpa, Juliana de França
Orientador(a): Rocha, Maria Valderez Ponte
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/21442
Resumo: Xylose reductase enzyme (EC 1.1.1.21) has potential application in the production of xylitol, and is an intracellular enzyme commonly found in yeast. Xylitol is a carbohydrate-alcohol used in food, dental, pharmaceutical and cosmetic industries. The large-scale production of this enzyme, as well as its industrial application for the production of xylitol and other bioproducts, has been limited due to the high price of commercial xylose. This problem has motivated researchers to develop better techniques for reducing the costs of obtaining XR, for example, the search for new raw material and the process conditions. Among possible feedstocks, lignocellulosic materials, as cashew apple bagasse, are low cost sources with potential applications in bioprocesses. In this context, the aim of this study was to evaluate the production of xylose reductase enzyme (XR) and xylitol by biotechnological processes by the yeast Candida tropicalis ATCC750 and hemicellulose hydrolysate from cashew apple bagasse (CAB) as substrate. The hemicellulose hydrolysate from cashew apple bagasse (HBCD) was obtained by acid hydrolysis with diluted sulfuric acid (H2SO4) and it used as a culture medium. The processes were conducted in orbital shaker at 150 rpm and different temperatures (25 °C, 30 °C, 35 °C and 45 °C). The xylose reductase enzyme extracts were used to reduce xylose to xylitol and the XR activity was determined by NADPH oxidation reaction at 25 °C. According to the analyzed data using hemicellulose hydrolysate (HBCD) medium, the highest microbial growth was observed at 25 °C and 30 °C and slightly growth at 40 °C. Under the experimental conditions evaluated, the production of xylitol was not observed, but there were the production of xylose reductase enzyme and ethanol. The XR from Candida tropicalis resulted in a crude extract with a higher enzymatic activity in the temperature of production at 25 °C (0.265 U/mL). In the partial characterization of the enzyme, the optimum pH and temperature were 7.0 and 50 °C, respectively. Subsequently, the gel electrophoresis was performed under denaturing conditions for identification of the enzyme fractions, which identify a heterodimeric structure with a molecular weight of approximately 30 kDa. The results showed that the cashew apple bagasse hydrolysate did not favor the xylitol production in the studied conditions; however, it is a potential means for biotechnological production of the enzyme xylose reductase (XR).
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spelling Serpa, Juliana de FrançaRocha, Maria Valderez Ponte2016-12-26T14:16:48Z2016-12-26T14:16:48Z2016SERPA, J. F. Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju. 2016. 68 f, Dissertação (Mestrado em Engenharia Química)–Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2016.http://www.repositorio.ufc.br/handle/riufc/21442Xylose reductase enzyme (EC 1.1.1.21) has potential application in the production of xylitol, and is an intracellular enzyme commonly found in yeast. Xylitol is a carbohydrate-alcohol used in food, dental, pharmaceutical and cosmetic industries. The large-scale production of this enzyme, as well as its industrial application for the production of xylitol and other bioproducts, has been limited due to the high price of commercial xylose. This problem has motivated researchers to develop better techniques for reducing the costs of obtaining XR, for example, the search for new raw material and the process conditions. Among possible feedstocks, lignocellulosic materials, as cashew apple bagasse, are low cost sources with potential applications in bioprocesses. In this context, the aim of this study was to evaluate the production of xylose reductase enzyme (XR) and xylitol by biotechnological processes by the yeast Candida tropicalis ATCC750 and hemicellulose hydrolysate from cashew apple bagasse (CAB) as substrate. The hemicellulose hydrolysate from cashew apple bagasse (HBCD) was obtained by acid hydrolysis with diluted sulfuric acid (H2SO4) and it used as a culture medium. The processes were conducted in orbital shaker at 150 rpm and different temperatures (25 °C, 30 °C, 35 °C and 45 °C). The xylose reductase enzyme extracts were used to reduce xylose to xylitol and the XR activity was determined by NADPH oxidation reaction at 25 °C. According to the analyzed data using hemicellulose hydrolysate (HBCD) medium, the highest microbial growth was observed at 25 °C and 30 °C and slightly growth at 40 °C. Under the experimental conditions evaluated, the production of xylitol was not observed, but there were the production of xylose reductase enzyme and ethanol. The XR from Candida tropicalis resulted in a crude extract with a higher enzymatic activity in the temperature of production at 25 °C (0.265 U/mL). In the partial characterization of the enzyme, the optimum pH and temperature were 7.0 and 50 °C, respectively. Subsequently, the gel electrophoresis was performed under denaturing conditions for identification of the enzyme fractions, which identify a heterodimeric structure with a molecular weight of approximately 30 kDa. The results showed that the cashew apple bagasse hydrolysate did not favor the xylitol production in the studied conditions; however, it is a potential means for biotechnological production of the enzyme xylose reductase (XR).A enzima xilose-redutase (EC 1.1.1.21) tem potencial aplicação na produção de xilitol a partir de xilose, e é uma enzima intracelular comumente encontrada em leveduras. O xilitol é um açúcar-álcool utilizado em indústrias alimentícia, odontológica, farmacêutica e de cosmético. A produção em larga escala desta enzima, bem como a sua aplicação industrial para a fabricação de xilitol e outros bioprodutos de valor agregado, tem sido limitado devido ao alto preço da xilose comercial. Este problema tem incentivado aos pesquisadores a trabalhar para o desenvolvimento de melhores técnicas para reduzir os custos de obtenção da XR, como, por exemplo, a busca por novas fontes de matéria-prima e condições do processo. Entre as possíveis matérias-primas, os materiais lignocelulósicos como, o bagaço de caju, são fontes de baixo custo com potenciais aplicações em bioprocessos. Neste contexto, o objetivo deste estudo foi avaliar a produção da enzima xilose redutase (XR) e de xilitol por processos biotecnológicos, através da levedura Candida tropicalis ATCC750 utilizando o hidrolisado hemicelulósico de bagaço de caju (BC) como substrato. O hidrolisado hemicelulósico do bagaço de caju (HBCD) foi obtido por hidrólise com ácido sulfúrico (H2SO4) diluído e aplicado como meio de cultivo. A produção da enzima foi realizado por bioprocessos conduzidos em agitador orbital a 150 rpm a diferentes temperaturas (25 °C, 30 °C, 35 °C e 45 °C). O extrato enzimático produzido foi utilizado para reduzir a xilose em xilitol e determinou-se a atividade da XR através da reação de oxidação de NADPH a 25 °C. De acordo com os dados analisados, o maior crescimento microbiano foi observado nas temperaturas de 25 °C e 30 °C, e diminuiu consideravelmente a 40 °C usando o meio hidrolisado hemicelulósico do bagaço de caju (HBCD). Nas condições avaliadas, não houve a produção de xilitol ocorrendo, no entanto, a produção da enzima xilose redutase e de etanol. A XR da Candida tropicalis resultou em um extrato bruto com uma maior atividade enzimática na temperatura de produção de 25 °C (0,265 U/mL). Na caracterização parcial da enzima, pH e a temperatura ótima da atividade enzimática foram pH 7,0 e 50 °C, respectivamente. Posteriormente, realizou-se a eletroforese em gel de poliacrilamida em condições desnaturantes para a identificação das frações da enzima, obtendo uma indicação de uma estrutura dimérica com peso molecular de, aproximadamente, 30 KDa. Com os resultados apresentados, pode-se considerar que o hidrolisado de bagaço de caju não favoreceu a produção de xilitol, contudo, é um meio potencial para a produção biotecnológica da enzima xilose redutase (XR).Engenharia químicaCandida tropicalisEnzimas - ProduçãoProdução da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de cajuProduction of xylose reductase enzyme by candida tropicalis ATCC750 using hemicellulose hydrolyzate from cashew apple bagasseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2016_dis_jfserpa.pdf2016_dis_jfserpa.pdfapplication/pdf1668343http://repositorio.ufc.br/bitstream/riufc/21442/1/2016_dis_jfserpa.pdf23472e000e232c0bab4b717ec91b621aMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/21442/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/214422020-09-17 11:46:32.368oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2020-09-17T14:46:32Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
dc.title.en.pt_BR.fl_str_mv Production of xylose reductase enzyme by candida tropicalis ATCC750 using hemicellulose hydrolyzate from cashew apple bagasse
title Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
spellingShingle Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
Serpa, Juliana de França
Engenharia química
Candida tropicalis
Enzimas - Produção
title_short Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
title_full Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
title_fullStr Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
title_full_unstemmed Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
title_sort Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju
author Serpa, Juliana de França
author_facet Serpa, Juliana de França
author_role author
dc.contributor.author.fl_str_mv Serpa, Juliana de França
dc.contributor.advisor1.fl_str_mv Rocha, Maria Valderez Ponte
contributor_str_mv Rocha, Maria Valderez Ponte
dc.subject.por.fl_str_mv Engenharia química
Candida tropicalis
Enzimas - Produção
topic Engenharia química
Candida tropicalis
Enzimas - Produção
description Xylose reductase enzyme (EC 1.1.1.21) has potential application in the production of xylitol, and is an intracellular enzyme commonly found in yeast. Xylitol is a carbohydrate-alcohol used in food, dental, pharmaceutical and cosmetic industries. The large-scale production of this enzyme, as well as its industrial application for the production of xylitol and other bioproducts, has been limited due to the high price of commercial xylose. This problem has motivated researchers to develop better techniques for reducing the costs of obtaining XR, for example, the search for new raw material and the process conditions. Among possible feedstocks, lignocellulosic materials, as cashew apple bagasse, are low cost sources with potential applications in bioprocesses. In this context, the aim of this study was to evaluate the production of xylose reductase enzyme (XR) and xylitol by biotechnological processes by the yeast Candida tropicalis ATCC750 and hemicellulose hydrolysate from cashew apple bagasse (CAB) as substrate. The hemicellulose hydrolysate from cashew apple bagasse (HBCD) was obtained by acid hydrolysis with diluted sulfuric acid (H2SO4) and it used as a culture medium. The processes were conducted in orbital shaker at 150 rpm and different temperatures (25 °C, 30 °C, 35 °C and 45 °C). The xylose reductase enzyme extracts were used to reduce xylose to xylitol and the XR activity was determined by NADPH oxidation reaction at 25 °C. According to the analyzed data using hemicellulose hydrolysate (HBCD) medium, the highest microbial growth was observed at 25 °C and 30 °C and slightly growth at 40 °C. Under the experimental conditions evaluated, the production of xylitol was not observed, but there were the production of xylose reductase enzyme and ethanol. The XR from Candida tropicalis resulted in a crude extract with a higher enzymatic activity in the temperature of production at 25 °C (0.265 U/mL). In the partial characterization of the enzyme, the optimum pH and temperature were 7.0 and 50 °C, respectively. Subsequently, the gel electrophoresis was performed under denaturing conditions for identification of the enzyme fractions, which identify a heterodimeric structure with a molecular weight of approximately 30 kDa. The results showed that the cashew apple bagasse hydrolysate did not favor the xylitol production in the studied conditions; however, it is a potential means for biotechnological production of the enzyme xylose reductase (XR).
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-12-26T14:16:48Z
dc.date.available.fl_str_mv 2016-12-26T14:16:48Z
dc.date.issued.fl_str_mv 2016
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dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv SERPA, J. F. Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju. 2016. 68 f, Dissertação (Mestrado em Engenharia Química)–Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2016.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/21442
identifier_str_mv SERPA, J. F. Produção da enzima xilose redutase por Candida tropicalis ATCC750 usando hidrolisado hemicelulósico do bagaço de caju. 2016. 68 f, Dissertação (Mestrado em Engenharia Química)–Centro de Tecnologia, Universidade Federal do Ceará, Fortaleza, 2016.
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