Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato

Detalhes bibliográficos
Ano de defesa: 2013
Autor(a) principal: Aquino, Patrícia Marina de
Orientador(a): Giordano, Raquel de Lima Camargo lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de São Carlos
Programa de Pós-Graduação: Programa de Pós-Graduação em Engenharia Química - PPGEQ
Departamento: Não Informado pela instituição
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: https://repositorio.ufscar.br/handle/ufscar/4125
Resumo: In this work, it was studied the simultaneous isomerization and fermentation of xylose to ethanol (SIF) using xylose isomerase (XI) and S. cerevisiae co-immobilized in calcium alginate gel. XI was immobilized on chitosan gel activated with glutaraldehyde (IXI-Ch). The influence of the concentration of enzyme/yeast in the reactor, the pH, temperature and yeast strain on yield and selectivity in ethanol was studied. The concentrations of enzyme and yeast in the reactor were varied by changing the mass of IXI-Ch and yeast per gram of alginate solution, maintaining fixed the ratios of biocatalyst weight: volume of medium in the reactor (1:1). The SIFs were carried out in batch with xylose (65g.L-1), antibiotics and other salts. The first experiment, with 16% Itaiquara® yeast and 5% enzyme biocatalyst (% wenzyme or yeast/wbiocatalyst) showed that pH drop occurred during the test, preventing full conversion of xylose, due to reduced enzyme activity. calcium carbonate (0.5-1.0%) was then included in the biocatalyst, which maintained the pH between 5,2 to 5,6, allowing complete conversion of the sugar at all concentrations tested (%Yeast -Enzyme in biocatalyst): 5-20, 17-5, and 10 yeast (Itaiquara ®) with 5, 10 and 20-. The maximum ethanol productivity, 2,44 ± 0,26g.L-1.h- 1 was obtained for the highest cell concentration and the highest selectivity ethanol/xylitol, 2,57 ± 0.4 and 2,42 ± 0,01 for the highest enzyme concentrations (10 and 20% with 10% yeast). These results indicated that the highest concentration of xylulose favored more selectivity to ethanol. Fermentation was then performed using no enzyme in biocatalyst with a prior isomerized syrup concentrated in xylulose containing 58g.L-1 xylulose and 9g.L-1 xylose and another with xylose only. At first, xylulose was completely assimilated in 5 hours, xylose was barely consumed in both assays, and ethanol selectivity was lower than that obtained in the SIF tests. Xylitol show thus to be produced mainly from xylulose and selectivity contrary to expectations did not directly increase with increasing xylulose concentration, indicating that the formation of ethanol/xylitol depends not only on external xylulose, and it is probably finely regulated in yeast. The concentrations of enzyme and yeast 20 and 10% (equivalent to 100gderived.L-1 reactor and 50gwd.L-1 reactor) were selected as the best, which were used to study the influence of pH and temperature, and also different strains. The increase of initial pH from 5.6 to 6.5 did not improve the productivity, yield, neither selectivity in ethanol. Temperatures tested for Itaiquara ® were 32, 35 and 37 ° C, and for industrial strains CAT-1 and BG-1: 32, 37 and 40 ° C. Viability remained above 90% for all assays at 24 hours. All three strains showed increased selectivity in ethanol with temperature reduction, obtaining the maximum selectivity for industrial strains (3,06 ± 0,24 - CAT-1 and 3,19 ± 0,11 BG-1) with yield and productivity equal or greater than those obtained in higher temperatures. At 32 ° C and pH 5.6, Itaiquara ® showed lower conversion time, but lower selectivity, while the BG-1, demonstrated the highest selectivity, but low conversion and productivity. The strain CAT-1 combines high productivity, 2,17 ± 0,17 g.L-1.h-1, and selectivity, 3,06 ± 0,24, with 90% conversion in 9 hours, 32 ° C, which is apparently the best performance among the tested yeasts. The results were very promising, indicating the technical feasibility of producing ethanol from xylose with the biocatalyst developed.
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spelling Aquino, Patrícia Marina deGiordano, Raquel de Lima Camargohttp://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4780181P0http://lattes.cnpq.br/60038058403771882016-06-02T19:56:52Z2013-09-042016-06-02T19:56:52Z2013-06-20https://repositorio.ufscar.br/handle/ufscar/4125In this work, it was studied the simultaneous isomerization and fermentation of xylose to ethanol (SIF) using xylose isomerase (XI) and S. cerevisiae co-immobilized in calcium alginate gel. XI was immobilized on chitosan gel activated with glutaraldehyde (IXI-Ch). The influence of the concentration of enzyme/yeast in the reactor, the pH, temperature and yeast strain on yield and selectivity in ethanol was studied. The concentrations of enzyme and yeast in the reactor were varied by changing the mass of IXI-Ch and yeast per gram of alginate solution, maintaining fixed the ratios of biocatalyst weight: volume of medium in the reactor (1:1). The SIFs were carried out in batch with xylose (65g.L-1), antibiotics and other salts. The first experiment, with 16% Itaiquara® yeast and 5% enzyme biocatalyst (% wenzyme or yeast/wbiocatalyst) showed that pH drop occurred during the test, preventing full conversion of xylose, due to reduced enzyme activity. calcium carbonate (0.5-1.0%) was then included in the biocatalyst, which maintained the pH between 5,2 to 5,6, allowing complete conversion of the sugar at all concentrations tested (%Yeast -Enzyme in biocatalyst): 5-20, 17-5, and 10 yeast (Itaiquara ®) with 5, 10 and 20-. The maximum ethanol productivity, 2,44 ± 0,26g.L-1.h- 1 was obtained for the highest cell concentration and the highest selectivity ethanol/xylitol, 2,57 ± 0.4 and 2,42 ± 0,01 for the highest enzyme concentrations (10 and 20% with 10% yeast). These results indicated that the highest concentration of xylulose favored more selectivity to ethanol. Fermentation was then performed using no enzyme in biocatalyst with a prior isomerized syrup concentrated in xylulose containing 58g.L-1 xylulose and 9g.L-1 xylose and another with xylose only. At first, xylulose was completely assimilated in 5 hours, xylose was barely consumed in both assays, and ethanol selectivity was lower than that obtained in the SIF tests. Xylitol show thus to be produced mainly from xylulose and selectivity contrary to expectations did not directly increase with increasing xylulose concentration, indicating that the formation of ethanol/xylitol depends not only on external xylulose, and it is probably finely regulated in yeast. The concentrations of enzyme and yeast 20 and 10% (equivalent to 100gderived.L-1 reactor and 50gwd.L-1 reactor) were selected as the best, which were used to study the influence of pH and temperature, and also different strains. The increase of initial pH from 5.6 to 6.5 did not improve the productivity, yield, neither selectivity in ethanol. Temperatures tested for Itaiquara ® were 32, 35 and 37 ° C, and for industrial strains CAT-1 and BG-1: 32, 37 and 40 ° C. Viability remained above 90% for all assays at 24 hours. All three strains showed increased selectivity in ethanol with temperature reduction, obtaining the maximum selectivity for industrial strains (3,06 ± 0,24 - CAT-1 and 3,19 ± 0,11 BG-1) with yield and productivity equal or greater than those obtained in higher temperatures. At 32 ° C and pH 5.6, Itaiquara ® showed lower conversion time, but lower selectivity, while the BG-1, demonstrated the highest selectivity, but low conversion and productivity. The strain CAT-1 combines high productivity, 2,17 ± 0,17 g.L-1.h-1, and selectivity, 3,06 ± 0,24, with 90% conversion in 9 hours, 32 ° C, which is apparently the best performance among the tested yeasts. The results were very promising, indicating the technical feasibility of producing ethanol from xylose with the biocatalyst developed.Neste trabalho foi estudada a simultânea isomerização e fermentação de xilose a etanol (SIF) usando xilose isomerase (XI) e S. cerevisiae coimobilizadas em gel de alginato de cálcio. XI foi imobilizada em gel de quitosana ativado com glutaraldeido (IXI-Ch).. Foram estudadas as influências das concentrações de enzima/levedura no reator, do pH, da temperatura e da linhagem de levedura na produtividade e na seletividade em etanol. As concentrações de enzima e levedura no reator foram variadas mudando-se a massa de IXI-Ch e levedura por grama de solução de alginato, mantendo-se fixas as proporções 1:1 massa de biocatalisador:volume de meio no reator. As SIFs foram realizadas em batelada com xilose (~65g.L-1), antibiótico e outros sais. O primeiro experimento realizado, biocatalisador com 16% levedura Itaiquara® e 5% enzima (% menzima ou levedura/mbiocatalisador), mostrou que ocorria queda de pH durante o ensaio, impedindo conversão total da xilose, devido à redução da atividade enzimática. Foi incluído carbonato de cálcio 0,5-1,0% no biocatalisador, o que manteve o pH entre 5,2-5,6, permitindo total conversão do açúcar, em todas as concentrações testadas (%Levedura-Enzima no biocatalisador): 5-20, 17-5 e 10 levedura (Itaiquara®)com 5, 10 e 20% enzima. A máxima produtividade em etanol, 2,44 ± 0,26g.L-1.h-1, foi obtida para a mais alta concentração celular e a mais alta seletividade etanol/xilitol, 2,57± 0,4 e 2,42± 0,01, para as mais altas concentrações de enzima (10 e 20% com 10% levedura). Esses resultados indicavam que quanto mais alta a concentração de xilulose, mais favorecida a seletividade em etanol. Foi então realizada uma fermentação usando o biocatalisador sem enzima, com um xarope previamente isomerizado e concentrado em xilulose contendo 58g/L de xilulose e 9g/L xilose e outro apenas com xilose. No primeiro, xilulose foi totalmente assimilada em 5 horas, xilose foi pouco consumida nos dois ensaios, e a seletividade em etanol foi menor que a obtida nos ensaios SIF. Xilitol mostrou, assim, ser produzido majoritariamente a partir de xilulose e contrariamente ao esperado a seletividade não aumenta diretamente com o aumento da concentração de xilulose, indicando que o metabolismo etanol/xilitol não depende apenas da concentração externa de xilulose, devendo ser finamente regulado dentro da levedura. Selecionaram-se as concentrações de enzima e levedura de 20 e 10% (equivalente a 100gderivado.L-1 reator e 50gms.L-1 reator) como as melhores, as quais foram utilizadas para estudo da influência do pH e da temperatura e ainda de diferentes linhagens. O aumento do pH inicial do meio de 5,6 para 6,5 não favoreceu a produtividade, rendimento e nem a seletividade em etanol. As temperaturas testadas para Itaiquara® foram: 32, 35 e 37°C; e para as linhagens industriais CAT-1 e BG-1: 32, 37 e 40°C. A viabilidade manteve-se acima de 90% para todos os ensaios em 24 horas. As três linhagens mostraram aumento da seletividade em etanol com a redução da temperatura, obtendo-se a máxima seletividade para as linhagens industriais (3,06± 0,24- CAT-1 e 3,19± 0,11 BG-1), com rendimento e produtividade iguais ou maiores que os obtidos nas temperaturas maiores. A 32°C e pH 5,6, Itaiquara® apresentou menor tempo de conversão, mas a menor seletividade, já a BG-1, obteve maior seletividade, mas baixa conversão, rendimento e produtividade. A linhagem CAT-1 alia alta produtividade, 2,17 ± 0,17 (g.L-1.h-1), e seletividade, 3,06 ± 0,24, com 90% de conversão em 9 horas, 32°C, sendo aparentemente a de melhor desempenho dentre as testadas. Os resultados foram muito promissores, indicando viabilidade técnica de produção de etanol a partir de xilose com o biocatalisador desenvolvido.Universidade Federal de Minas Geraisapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Engenharia Química - PPGEQUFSCarBRFermentaçãoIsomerizaçãoXiloseXilose isomeraseSaccharomyces cerevisiaeSimultânea isomerização e fermentação (SIF), EtanolSimultaneous isomerization and fermentation (SIF)EthanolENGENHARIAS::ENGENHARIA QUIMICAProdução de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginatoEthanol Production from Xylose with xylose isomerase and Saccharomyces cerevisiae co-immobilized alginate gelinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL5338.pdfapplication/pdf2812584https://{{ getenv "DSPACE_HOST" "repositorio.ufscar.br" }}/bitstream/ufscar/4125/1/5338.pdfc4ea2f65f591bec0e6ad7105c6ec591eMD51TEXT5338.pdf.txt5338.pdf.txtExtracted texttext/plain0https://{{ getenv "DSPACE_HOST" "repositorio.ufscar.br" }}/bitstream/ufscar/4125/2/5338.pdf.txtd41d8cd98f00b204e9800998ecf8427eMD52THUMBNAIL5338.pdf.jpg5338.pdf.jpgIM Thumbnailimage/jpeg6866https://{{ getenv "DSPACE_HOST" "repositorio.ufscar.br" }}/bitstream/ufscar/4125/3/5338.pdf.jpg2a8b6d351bbb5afa8b9834ea5ac7c42fMD53ufscar/41252019-09-11 04:13:05.661oai:repositorio.ufscar.br:ufscar/4125Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-05-25T12:48:14.476712Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
dc.title.alternative.eng.fl_str_mv Ethanol Production from Xylose with xylose isomerase and Saccharomyces cerevisiae co-immobilized alginate gel
title Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
spellingShingle Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
Aquino, Patrícia Marina de
Fermentação
Isomerização
Xilose
Xilose isomerase
Saccharomyces cerevisiae
Simultânea isomerização e fermentação (SIF), Etanol
Simultaneous isomerization and fermentation (SIF)
Ethanol
ENGENHARIAS::ENGENHARIA QUIMICA
title_short Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
title_full Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
title_fullStr Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
title_full_unstemmed Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
title_sort Produção de etanol a partir de xilose com glicose isomerase e Saccharomyces cerevisiae coimobilizadas em gel de alginato
author Aquino, Patrícia Marina de
author_facet Aquino, Patrícia Marina de
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/6003805840377188
dc.contributor.author.fl_str_mv Aquino, Patrícia Marina de
dc.contributor.advisor1.fl_str_mv Giordano, Raquel de Lima Camargo
dc.contributor.advisor1Lattes.fl_str_mv http://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4780181P0
contributor_str_mv Giordano, Raquel de Lima Camargo
dc.subject.por.fl_str_mv Fermentação
Isomerização
Xilose
Xilose isomerase
Saccharomyces cerevisiae
Simultânea isomerização e fermentação (SIF), Etanol
topic Fermentação
Isomerização
Xilose
Xilose isomerase
Saccharomyces cerevisiae
Simultânea isomerização e fermentação (SIF), Etanol
Simultaneous isomerization and fermentation (SIF)
Ethanol
ENGENHARIAS::ENGENHARIA QUIMICA
dc.subject.eng.fl_str_mv Simultaneous isomerization and fermentation (SIF)
Ethanol
dc.subject.cnpq.fl_str_mv ENGENHARIAS::ENGENHARIA QUIMICA
description In this work, it was studied the simultaneous isomerization and fermentation of xylose to ethanol (SIF) using xylose isomerase (XI) and S. cerevisiae co-immobilized in calcium alginate gel. XI was immobilized on chitosan gel activated with glutaraldehyde (IXI-Ch). The influence of the concentration of enzyme/yeast in the reactor, the pH, temperature and yeast strain on yield and selectivity in ethanol was studied. The concentrations of enzyme and yeast in the reactor were varied by changing the mass of IXI-Ch and yeast per gram of alginate solution, maintaining fixed the ratios of biocatalyst weight: volume of medium in the reactor (1:1). The SIFs were carried out in batch with xylose (65g.L-1), antibiotics and other salts. The first experiment, with 16% Itaiquara® yeast and 5% enzyme biocatalyst (% wenzyme or yeast/wbiocatalyst) showed that pH drop occurred during the test, preventing full conversion of xylose, due to reduced enzyme activity. calcium carbonate (0.5-1.0%) was then included in the biocatalyst, which maintained the pH between 5,2 to 5,6, allowing complete conversion of the sugar at all concentrations tested (%Yeast -Enzyme in biocatalyst): 5-20, 17-5, and 10 yeast (Itaiquara ®) with 5, 10 and 20-. The maximum ethanol productivity, 2,44 ± 0,26g.L-1.h- 1 was obtained for the highest cell concentration and the highest selectivity ethanol/xylitol, 2,57 ± 0.4 and 2,42 ± 0,01 for the highest enzyme concentrations (10 and 20% with 10% yeast). These results indicated that the highest concentration of xylulose favored more selectivity to ethanol. Fermentation was then performed using no enzyme in biocatalyst with a prior isomerized syrup concentrated in xylulose containing 58g.L-1 xylulose and 9g.L-1 xylose and another with xylose only. At first, xylulose was completely assimilated in 5 hours, xylose was barely consumed in both assays, and ethanol selectivity was lower than that obtained in the SIF tests. Xylitol show thus to be produced mainly from xylulose and selectivity contrary to expectations did not directly increase with increasing xylulose concentration, indicating that the formation of ethanol/xylitol depends not only on external xylulose, and it is probably finely regulated in yeast. The concentrations of enzyme and yeast 20 and 10% (equivalent to 100gderived.L-1 reactor and 50gwd.L-1 reactor) were selected as the best, which were used to study the influence of pH and temperature, and also different strains. The increase of initial pH from 5.6 to 6.5 did not improve the productivity, yield, neither selectivity in ethanol. Temperatures tested for Itaiquara ® were 32, 35 and 37 ° C, and for industrial strains CAT-1 and BG-1: 32, 37 and 40 ° C. Viability remained above 90% for all assays at 24 hours. All three strains showed increased selectivity in ethanol with temperature reduction, obtaining the maximum selectivity for industrial strains (3,06 ± 0,24 - CAT-1 and 3,19 ± 0,11 BG-1) with yield and productivity equal or greater than those obtained in higher temperatures. At 32 ° C and pH 5.6, Itaiquara ® showed lower conversion time, but lower selectivity, while the BG-1, demonstrated the highest selectivity, but low conversion and productivity. The strain CAT-1 combines high productivity, 2,17 ± 0,17 g.L-1.h-1, and selectivity, 3,06 ± 0,24, with 90% conversion in 9 hours, 32 ° C, which is apparently the best performance among the tested yeasts. The results were very promising, indicating the technical feasibility of producing ethanol from xylose with the biocatalyst developed.
publishDate 2013
dc.date.available.fl_str_mv 2013-09-04
2016-06-02T19:56:52Z
dc.date.issued.fl_str_mv 2013-06-20
dc.date.accessioned.fl_str_mv 2016-06-02T19:56:52Z
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dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Engenharia Química - PPGEQ
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