Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium
Ano de defesa: | 2017 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de São Carlos
Câmpus São Carlos |
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Engenharia Química - PPGEQ
|
Departamento: |
Não Informado pela instituição
|
País: |
Não Informado pela instituição
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | https://repositorio.ufscar.br/handle/ufscar/8888 |
Resumo: | In recent years, the application of attenuated Salmonella spp. has been investigated for development of several biotechnological products, mainly vaccines. However, the implementation of industrial processes to obtain these products depends on the development of strategies for this microorganism high-cell density cultures (HCDC). One of the HCDC’s greatest challenges is overcoming Salmonella’s metabolic limitations, as it presents a high organic acids production (mainly acetic acid) that inhibits biomass formation. In this context, this work proposes two approaches to deal with this problem and implement Salmonella’s HCDC: studying the cultivation of S. typhimurium using glycerol as carbon source to reduce the generation of organic acids; studying the cultivation of a recombinant strain of S. typhimurium expressing the enzyme acetyl-CoA synthetase (ACS) from E. coli, for improved acetate assimilation). Initially, cultures were grown in agitated flasks in minimal media for two carbon sources (glucose or glycerol) for the wild-type and the recombinant strain. After the preliminary experiments, the recombinant strain was cultivated in bioreactor operated in batch mode with minimal medium formulated with glycerol, glucose or acetic acid as carbon source, to evaluate acetate production and assimilation. The wild-type strain was cultivated in a continuous-mode bioreactor on minimal medium with glycerol at D=0.10; 0.17 and 0.22 h-1 to evaluate the S. typhimurium glycerol metabolism. During the cultivations, samples were collected and analyzed by high-performance liquid chromatography to quantify the production of organic acids and substrate consumption. To quantify the concentration of biomass, optical density (600 nm) readings of culture broth and dry cell weight measurements were performed. Agitated flasks and batch cultivations results indicated that the acetate production is reduced in medium with glycerol for both strains, and that the genetically modified cells present a lower acetate accumulation phenotype compared to the wild-type. Continuous cultures of the wild-type strain showed no acetate accumulation for a 0.1 h-1 dilution rate. At rates of 0.17 h-1 and 0.22 h-1 acetate accumulation was observed, but acetate flux was 2-fold lower than the flux reached in chemostat with glucose-formulated medium. Simulations were performed with the STM_v1.0 model using as input data the glycerol and oxygen fluxes estimated from the experimental results. Good predictions were obtained for the biomass, CO2 and acetate fluxes at the higher dilution rates. The results suggest that fed-batch culture using glycerol as carbon source along with an exponential feed to maintain a 0.1 h-1 specific growth rate as a promising strategy to obtain high cellular concentrations of wild-type S. typhimurium. The efficient acetate uptake observed for the recombinant S. typhimurium cells may allow higher values of specific growth rate to be used for this strain, resulting in a higher productivity of biomass. |
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Fuzer Neto, José RobertoSilva, Adilson José dahttp://lattes.cnpq.br/3447469350644179Zangirolami, Teresa Cristinahttp://lattes.cnpq.br/4546701843297248http://lattes.cnpq.br/65433293937583912f1d5ee5-5569-42d5-8027-d615d60598612017-06-28T08:24:37Z2017-06-28T08:24:37Z2017-02-23FUZER NETO, José Roberto. Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium. 2017. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2017. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8888.https://repositorio.ufscar.br/handle/ufscar/8888In recent years, the application of attenuated Salmonella spp. has been investigated for development of several biotechnological products, mainly vaccines. However, the implementation of industrial processes to obtain these products depends on the development of strategies for this microorganism high-cell density cultures (HCDC). One of the HCDC’s greatest challenges is overcoming Salmonella’s metabolic limitations, as it presents a high organic acids production (mainly acetic acid) that inhibits biomass formation. In this context, this work proposes two approaches to deal with this problem and implement Salmonella’s HCDC: studying the cultivation of S. typhimurium using glycerol as carbon source to reduce the generation of organic acids; studying the cultivation of a recombinant strain of S. typhimurium expressing the enzyme acetyl-CoA synthetase (ACS) from E. coli, for improved acetate assimilation). Initially, cultures were grown in agitated flasks in minimal media for two carbon sources (glucose or glycerol) for the wild-type and the recombinant strain. After the preliminary experiments, the recombinant strain was cultivated in bioreactor operated in batch mode with minimal medium formulated with glycerol, glucose or acetic acid as carbon source, to evaluate acetate production and assimilation. The wild-type strain was cultivated in a continuous-mode bioreactor on minimal medium with glycerol at D=0.10; 0.17 and 0.22 h-1 to evaluate the S. typhimurium glycerol metabolism. During the cultivations, samples were collected and analyzed by high-performance liquid chromatography to quantify the production of organic acids and substrate consumption. To quantify the concentration of biomass, optical density (600 nm) readings of culture broth and dry cell weight measurements were performed. Agitated flasks and batch cultivations results indicated that the acetate production is reduced in medium with glycerol for both strains, and that the genetically modified cells present a lower acetate accumulation phenotype compared to the wild-type. Continuous cultures of the wild-type strain showed no acetate accumulation for a 0.1 h-1 dilution rate. At rates of 0.17 h-1 and 0.22 h-1 acetate accumulation was observed, but acetate flux was 2-fold lower than the flux reached in chemostat with glucose-formulated medium. Simulations were performed with the STM_v1.0 model using as input data the glycerol and oxygen fluxes estimated from the experimental results. Good predictions were obtained for the biomass, CO2 and acetate fluxes at the higher dilution rates. The results suggest that fed-batch culture using glycerol as carbon source along with an exponential feed to maintain a 0.1 h-1 specific growth rate as a promising strategy to obtain high cellular concentrations of wild-type S. typhimurium. The efficient acetate uptake observed for the recombinant S. typhimurium cells may allow higher values of specific growth rate to be used for this strain, resulting in a higher productivity of biomass.Nos últimos anos, a aplicação de linhagens atenuadas de Salmonella spp. vem sendo amplamente investigada para o desenvolvimento de diversos produtos biotecnológicos, principalmente vacinas. No entanto, a implementação de processos industriais para a obtenção destes produtos depende do desenvolvimento de estratégias para o cultivo em alta densidade celular (CADC) deste microrganismo. Para isso, um dos grandes desafios a ser superado se refere às limitações metabólicas da Salmonella, uma vez que esta apresenta elevada produção de ácidos orgânicos (principalmente ácido acético) que inibem a formação de biomassa. Neste contexto, este trabalho propõe duas abordagens para lidar com este problema e implementar CADC de Salmonella: estudar o crescimento de Salmonella typhimurium em glicerol avaliando seu metabolismo como uma fonte de carbono menos propensa à geração de ácidos orgânicos; e estudar a produção destes ácidos por uma cepa de S. typhimurium geneticamente modificada para diminuir o acúmulo de acetato (super-expressão do gene acs, de Escherichia coli, que codifica a enzima acetil-CoA sintetase (ACS) responsável pela assimilação de acetato). Inicialmente foram realizados cultivos em frascos agitados em meio mínimo para duas fontes de carbono (glicose ou glicerol), para a cepa selvagem e para a cepa recombinante. Após os experimentos preliminares, foram realizados cultivos em biorreator operado em modo batelada com a cepa modificada em meio mínimo formulado com glicerol, glicose ou ácido acético, como fonte de carbono, a fim de avaliar a produção e a assimilação de acetato. Foram realizados cultivos em biorreator em modo contínuo com a cepa selvagem em meio mínimo com glicerol com D=0,10; 0,17 e 0,22 h-1 para avaliar o metabolismo do glicerol pela S. typhimurium. Ao longo dos cultivos foram coletadas amostras do caldo e analisadas por cromatografia líquida de alta eficiência para quantificar a produção de ácidos orgânicos e o consumo de substrato. Para quantificar a biomassa produzida, foram realizadas medidas de densidade ótica (600 nm) e de massa seca. Os resultados dos cultivos em frascos agitados e das bateladas indicam que, para ambas as cepas, a produção de acetato é reduzida em meio formulado com glicerol, e que as células modificadas geneticamente apresentam um fenótipo de menor acúmulo de acetato comparadas à linhagem selvagem. Os cultivos contínuos realizados com a cepa selvagem mostraram que não houve acúmulo de acetato para a taxa de diluição de 0,1 h-1. Já nas taxas de 0,17 h-1 e 0,22 h-1, apesar de haver acúmulo, o fluxo de produção de acetato foi cerca de 2 vezes menor que o observado em quimiostato em meio formulado com glicose. Foram realizadas simulações com o modelo STM_v1.0, tendo como dados de entrada os fluxos de glicerol e de oxigênio estimados a partir dos dados experimentais. O modelo descreveu bem os fluxos de biomassa, CO2 e acetato para as taxas de diluição mais altas. Os resultados obtidos sugerem o cultivo em batelada alimentada com glicerol como fonte de carbono, e alimentação exponencial definida para manter a velocidade específica de crescimento em 0,1 h-1, como uma estratégia promissora para obter altas concentrações celulares de S. typhimurium selvagem. Para a S. typhimurium recombinante, devido à sua eficiente assimilação de acetato, valores ainda maiores de velocidade específica de crescimento poderiam ser impostos, com elevado aumento de produtividade em biomassa.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Engenharia Química - PPGEQUFSCarSalmonella typhimuriumCultivos em biorreatorQuimiostatoMetabolismo do carbono centralGlicerolBioreactor cultivationChemostatCentral carbon metabolismGlycerolENGENHARIAS::ENGENHARIA QUIMICAEstratégias para redução da produção de acetato em cultivos de Salmonella typhimuriuminfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline6006001c7296d8-a6a1-4938-92ff-1182b3437864info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDissJRFN.pdfDissJRFN.pdfapplication/pdf1477527https://repositorio.ufscar.br/bitstream/ufscar/8888/1/DissJRFN.pdfcb09da4bfdd14f1e2c076f57d365b7a1MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8888/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTDissJRFN.pdf.txtDissJRFN.pdf.txtExtracted texttext/plain167348https://repositorio.ufscar.br/bitstream/ufscar/8888/3/DissJRFN.pdf.txt39f694183b259b468d9ec12e7b524e5bMD53THUMBNAILDissJRFN.pdf.jpgDissJRFN.pdf.jpgIM Thumbnailimage/jpeg6013https://repositorio.ufscar.br/bitstream/ufscar/8888/4/DissJRFN.pdf.jpg33932e8fa60857607373386a79eeb244MD54ufscar/88882023-09-18 18:31:24.771oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:24Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
title |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
spellingShingle |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium Fuzer Neto, José Roberto Salmonella typhimurium Cultivos em biorreator Quimiostato Metabolismo do carbono central Glicerol Bioreactor cultivation Chemostat Central carbon metabolism Glycerol ENGENHARIAS::ENGENHARIA QUIMICA |
title_short |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
title_full |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
title_fullStr |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
title_full_unstemmed |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
title_sort |
Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium |
author |
Fuzer Neto, José Roberto |
author_facet |
Fuzer Neto, José Roberto |
author_role |
author |
dc.contributor.advisor.none.fl_str_mv |
|
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/6543329393758391 |
dc.contributor.author.fl_str_mv |
Fuzer Neto, José Roberto |
dc.contributor.advisor1.fl_str_mv |
Silva, Adilson José da |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3447469350644179 |
dc.contributor.advisor-co1.fl_str_mv |
Zangirolami, Teresa Cristina |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/4546701843297248 |
dc.contributor.authorID.fl_str_mv |
2f1d5ee5-5569-42d5-8027-d615d6059861 |
contributor_str_mv |
Silva, Adilson José da Zangirolami, Teresa Cristina |
dc.subject.por.fl_str_mv |
Salmonella typhimurium Cultivos em biorreator Quimiostato Metabolismo do carbono central Glicerol |
topic |
Salmonella typhimurium Cultivos em biorreator Quimiostato Metabolismo do carbono central Glicerol Bioreactor cultivation Chemostat Central carbon metabolism Glycerol ENGENHARIAS::ENGENHARIA QUIMICA |
dc.subject.eng.fl_str_mv |
Bioreactor cultivation Chemostat Central carbon metabolism Glycerol |
dc.subject.cnpq.fl_str_mv |
ENGENHARIAS::ENGENHARIA QUIMICA |
description |
In recent years, the application of attenuated Salmonella spp. has been investigated for development of several biotechnological products, mainly vaccines. However, the implementation of industrial processes to obtain these products depends on the development of strategies for this microorganism high-cell density cultures (HCDC). One of the HCDC’s greatest challenges is overcoming Salmonella’s metabolic limitations, as it presents a high organic acids production (mainly acetic acid) that inhibits biomass formation. In this context, this work proposes two approaches to deal with this problem and implement Salmonella’s HCDC: studying the cultivation of S. typhimurium using glycerol as carbon source to reduce the generation of organic acids; studying the cultivation of a recombinant strain of S. typhimurium expressing the enzyme acetyl-CoA synthetase (ACS) from E. coli, for improved acetate assimilation). Initially, cultures were grown in agitated flasks in minimal media for two carbon sources (glucose or glycerol) for the wild-type and the recombinant strain. After the preliminary experiments, the recombinant strain was cultivated in bioreactor operated in batch mode with minimal medium formulated with glycerol, glucose or acetic acid as carbon source, to evaluate acetate production and assimilation. The wild-type strain was cultivated in a continuous-mode bioreactor on minimal medium with glycerol at D=0.10; 0.17 and 0.22 h-1 to evaluate the S. typhimurium glycerol metabolism. During the cultivations, samples were collected and analyzed by high-performance liquid chromatography to quantify the production of organic acids and substrate consumption. To quantify the concentration of biomass, optical density (600 nm) readings of culture broth and dry cell weight measurements were performed. Agitated flasks and batch cultivations results indicated that the acetate production is reduced in medium with glycerol for both strains, and that the genetically modified cells present a lower acetate accumulation phenotype compared to the wild-type. Continuous cultures of the wild-type strain showed no acetate accumulation for a 0.1 h-1 dilution rate. At rates of 0.17 h-1 and 0.22 h-1 acetate accumulation was observed, but acetate flux was 2-fold lower than the flux reached in chemostat with glucose-formulated medium. Simulations were performed with the STM_v1.0 model using as input data the glycerol and oxygen fluxes estimated from the experimental results. Good predictions were obtained for the biomass, CO2 and acetate fluxes at the higher dilution rates. The results suggest that fed-batch culture using glycerol as carbon source along with an exponential feed to maintain a 0.1 h-1 specific growth rate as a promising strategy to obtain high cellular concentrations of wild-type S. typhimurium. The efficient acetate uptake observed for the recombinant S. typhimurium cells may allow higher values of specific growth rate to be used for this strain, resulting in a higher productivity of biomass. |
publishDate |
2017 |
dc.date.accessioned.fl_str_mv |
2017-06-28T08:24:37Z |
dc.date.available.fl_str_mv |
2017-06-28T08:24:37Z |
dc.date.issued.fl_str_mv |
2017-02-23 |
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.citation.fl_str_mv |
FUZER NETO, José Roberto. Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium. 2017. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2017. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8888. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/8888 |
identifier_str_mv |
FUZER NETO, José Roberto. Estratégias para redução da produção de acetato em cultivos de Salmonella typhimurium. 2017. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2017. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8888. |
url |
https://repositorio.ufscar.br/handle/ufscar/8888 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.confidence.fl_str_mv |
600 600 |
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1c7296d8-a6a1-4938-92ff-1182b3437864 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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 |
publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFSCAR instname:Universidade Federal de São Carlos (UFSCAR) instacron:UFSCAR |
instname_str |
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instacron_str |
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institution |
UFSCAR |
reponame_str |
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collection |
Repositório Institucional da UFSCAR |
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