Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz

Detalhes bibliográficos
Autor(a) principal: Leonel, Lillian Vieira
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações do UNIOESTE
Texto Completo: http://tede.unioeste.br/handle/tede/5020
Resumo: Rice husk is a waste derived from rice milling and is an abundant byproduct in cereal producing countries. Due to the considerable contents of cellulose in its composition, it is a potential raw material for use in various bioprocesses, such as ethanol production. However, several technical challenges must be overcome to make the second-generation ethanol production process economically viable and competitive. In this context, the process of simultaneous saccharification and fermentation (SSF), associated with the increase of solids concentration, as well as the use of surfactants in enzymatic hydrolysis in fermentation processes that employ lignocellulosic biomass are promising strategies to obtain higher sugar yields and consequently ethanol. Thus, the objective of this work was to evaluate different strategies to obtain an improvement in ethanol production from rice husk cellulose, using the thermotolerant yeast Kluyveromyces marxianus ATCC 36907. The first strategy was to carry out the fed batch SSF to increase the solids load; the second was to evaluate the effect of adding nonionic surfactants during enzymatic hydrolysis in the SSF process. First, rice husk biomass was subjected to chemical characterization for cellulose, hemicellulose, and lignin content, before and after alkaline pretreatment with 8% (w/v) sodium hydroxide (NaOH). The pretreated rice husk was subjected to enzymatic hydrolysis with different enzyme loading (5, 11, 22 FPU/gcelullose) and 10% solids (w/v) to determine the condition with increased release of reducing sugars for subsequent SSF use in batch fed with high solids loading. For the SSF in fed batch, four experiments were performed with different biomass concentrations of rice husk, being E1 and E2 experiments with initial load of 10% and final 20% (w/v) and E3 and E4 with initial load 15% and final 25% (w/v), respectively, at pH 5.5; 40 °C, 200 rpm, 96 hours. Thus, feeding intervals were established, with two feedings every 24 hours (E1 and E3) and four feedings every 12 hours (E2 and E4). As a second strategy, enzymatic hydrolysis was carried out, with the objective of evaluating the effect of different enzyme loads (5; 8; 13.5; 19 and 22 FPU/gcellulose) it was evaluated in association with Polysorbate 80 (PS80) surfactants (7.0, 10, 15, 20 and 23 mg/L), Polyethylene glycol 4000 (PEG) (0.5, 1.0, 2.0, 3.0 and 3.5 g/L) and Triton X-100 (TX-100) (75; 100; 150, 200 and 225 mg/L) in glucose release through three experiments following a central rotational composite design (DCCR) 22, including 4 axial conditions and 4 center point repetitions, totaling 12 assays for each surfactant, besides a control experiment (with enzyme addition and without surfactant), in triplicate. The third strategy consisted of adding the surfactant SSF, to evaluate the influence of the surfactant in the production of ethanol. For this, three experiments were performed: control (without surfactant + 19 FPU/gcellulose), with addition of Polysorbate 80 (10.0 mg/L + 19 FPU/gcellulose) and with Polyethylene Glycol 4000 (3.0 g/L + 19 FPU/gcellulose), the type of surfactant and respective concentrations and enzyme load selected from the results of enzymatic hydrolysis that were significant. The experiments were carried out with a solids ratio of 10% (m / v), pH 5.5, 40 °C, 200 rpm, 96 hours, in triplicate. The chemical composition of rice husk was 36.03% cellulose, 14.71% hemicellulose and 26.47% lignin and after pretreatment with 8% NaOH (w/v) the content of these fractions was 70.27; 8.39 and 21.34% cellulose, hemicellulose, and lignin, respectively. In the fed batch SSF with high solids loading, an ethanol production was obtained of 9.57; 9.92; 13.99 and 12.77 g/L, yields (YE/C) of 0.16; 0.17; 0.18 and 0.18 g/g and volumetric productivity (QP) of 0.27; 0.28; 0.39 and 0.53 g/L.h-1 for the experiments E1, E2, E3 and E4, respectively. For the enzymatic hydrolysis, glucose concentrations ranged from 32.36 to 79.57 g/L with PS80 addition, from 45.14 to 74.60 g/L with PEG and 32.28 to 47.89 g/L with addition of TX-100, while the control showed from 20.36 to 47.77 g/L in 96 hours. Only the ix surfactants PS80 and PEG in association with different enzyme loads promoted significant effects (p<0.05), the enzymatic load being the factor that most influenced the increase of final glucose production. Regarding SSF with surfactant, the addition of PS80 (10 mg/L) and PEG (3.0 g/L) in association with 19 FPU/gcellulose resulted in 6.73 and 8.17 g/L of ethanol, YE/C of 0.17 and 0.21 g / g and QP of 0.56 and 0.68 g/Lh-1 , in 12 hours of fermentation, respectively. The results suggest that rice husk is a raw material that can be used as a substrate in bioprocesses because it has a cellulose content similar to that of other lignocellulosic biomasses. The different rice husk feeding strategies used in fed batch SSF did not result in substantial increases in ethanol production. The addition of surfactants PS80 and PEG during enzymatic hydrolysis resulted in significant increases in final glucose production. In SSF, the same surfactants contributed to a slight increase in ethanol production in a shorter fermentation time, compared to the control.
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spelling Sene, Lucianehttp://lattes.cnpq.br/2582084888410031Arruda, Priscila Vaz dehttp://lattes.cnpq.br/1583339937667600Cunha, Mário Antônio Alves dahttp://lattes.cnpq.br/3151576713472624Kadowaki, Marina Kimikohttp://lattes.cnpq.br/1819723253019762Gomes, Simone Damascenohttp://lattes.cnpq.br/3362104483832351http://lattes.cnpq.br/9257508293525181Leonel, Lillian Vieira2020-10-09T18:37:45Z2020-03-12LEONEL, Lillian Vieira. Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz. 2020. 102 f. Tese (Programa de Pós-Graduação em Engenharia Agrícola) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.http://tede.unioeste.br/handle/tede/5020Rice husk is a waste derived from rice milling and is an abundant byproduct in cereal producing countries. Due to the considerable contents of cellulose in its composition, it is a potential raw material for use in various bioprocesses, such as ethanol production. However, several technical challenges must be overcome to make the second-generation ethanol production process economically viable and competitive. In this context, the process of simultaneous saccharification and fermentation (SSF), associated with the increase of solids concentration, as well as the use of surfactants in enzymatic hydrolysis in fermentation processes that employ lignocellulosic biomass are promising strategies to obtain higher sugar yields and consequently ethanol. Thus, the objective of this work was to evaluate different strategies to obtain an improvement in ethanol production from rice husk cellulose, using the thermotolerant yeast Kluyveromyces marxianus ATCC 36907. The first strategy was to carry out the fed batch SSF to increase the solids load; the second was to evaluate the effect of adding nonionic surfactants during enzymatic hydrolysis in the SSF process. First, rice husk biomass was subjected to chemical characterization for cellulose, hemicellulose, and lignin content, before and after alkaline pretreatment with 8% (w/v) sodium hydroxide (NaOH). The pretreated rice husk was subjected to enzymatic hydrolysis with different enzyme loading (5, 11, 22 FPU/gcelullose) and 10% solids (w/v) to determine the condition with increased release of reducing sugars for subsequent SSF use in batch fed with high solids loading. For the SSF in fed batch, four experiments were performed with different biomass concentrations of rice husk, being E1 and E2 experiments with initial load of 10% and final 20% (w/v) and E3 and E4 with initial load 15% and final 25% (w/v), respectively, at pH 5.5; 40 °C, 200 rpm, 96 hours. Thus, feeding intervals were established, with two feedings every 24 hours (E1 and E3) and four feedings every 12 hours (E2 and E4). As a second strategy, enzymatic hydrolysis was carried out, with the objective of evaluating the effect of different enzyme loads (5; 8; 13.5; 19 and 22 FPU/gcellulose) it was evaluated in association with Polysorbate 80 (PS80) surfactants (7.0, 10, 15, 20 and 23 mg/L), Polyethylene glycol 4000 (PEG) (0.5, 1.0, 2.0, 3.0 and 3.5 g/L) and Triton X-100 (TX-100) (75; 100; 150, 200 and 225 mg/L) in glucose release through three experiments following a central rotational composite design (DCCR) 22, including 4 axial conditions and 4 center point repetitions, totaling 12 assays for each surfactant, besides a control experiment (with enzyme addition and without surfactant), in triplicate. The third strategy consisted of adding the surfactant SSF, to evaluate the influence of the surfactant in the production of ethanol. For this, three experiments were performed: control (without surfactant + 19 FPU/gcellulose), with addition of Polysorbate 80 (10.0 mg/L + 19 FPU/gcellulose) and with Polyethylene Glycol 4000 (3.0 g/L + 19 FPU/gcellulose), the type of surfactant and respective concentrations and enzyme load selected from the results of enzymatic hydrolysis that were significant. The experiments were carried out with a solids ratio of 10% (m / v), pH 5.5, 40 °C, 200 rpm, 96 hours, in triplicate. The chemical composition of rice husk was 36.03% cellulose, 14.71% hemicellulose and 26.47% lignin and after pretreatment with 8% NaOH (w/v) the content of these fractions was 70.27; 8.39 and 21.34% cellulose, hemicellulose, and lignin, respectively. In the fed batch SSF with high solids loading, an ethanol production was obtained of 9.57; 9.92; 13.99 and 12.77 g/L, yields (YE/C) of 0.16; 0.17; 0.18 and 0.18 g/g and volumetric productivity (QP) of 0.27; 0.28; 0.39 and 0.53 g/L.h-1 for the experiments E1, E2, E3 and E4, respectively. For the enzymatic hydrolysis, glucose concentrations ranged from 32.36 to 79.57 g/L with PS80 addition, from 45.14 to 74.60 g/L with PEG and 32.28 to 47.89 g/L with addition of TX-100, while the control showed from 20.36 to 47.77 g/L in 96 hours. Only the ix surfactants PS80 and PEG in association with different enzyme loads promoted significant effects (p<0.05), the enzymatic load being the factor that most influenced the increase of final glucose production. Regarding SSF with surfactant, the addition of PS80 (10 mg/L) and PEG (3.0 g/L) in association with 19 FPU/gcellulose resulted in 6.73 and 8.17 g/L of ethanol, YE/C of 0.17 and 0.21 g / g and QP of 0.56 and 0.68 g/Lh-1 , in 12 hours of fermentation, respectively. The results suggest that rice husk is a raw material that can be used as a substrate in bioprocesses because it has a cellulose content similar to that of other lignocellulosic biomasses. The different rice husk feeding strategies used in fed batch SSF did not result in substantial increases in ethanol production. The addition of surfactants PS80 and PEG during enzymatic hydrolysis resulted in significant increases in final glucose production. In SSF, the same surfactants contributed to a slight increase in ethanol production in a shorter fermentation time, compared to the control.A casca de arroz é um resíduo proveniente do beneficiamento do arroz, sendo um subproduto abundante em países produtores do cereal. Por apresentar teores consideráveis de celulose em sua composição, é uma potencial matéria-prima para utilização em vários bioprocessos, como a produção de etanol. No entanto, vários desafios técnicos precisam ser superados a fim de tornar o processo de produção de etanol de segunda geração economicamente viável e competitivo. Nesse contexto, o processo de sacarificação, simultânea à fermentação (SSF), associado ao aumento da concentração de sólidos, bem como à utilização de surfactantes na hidrólise enzimática, em processos fermentativos, que empregam biomassa lignocelulósica, são estratégias promissoras para a obtenção de maiores rendimentos de açúcares e, consequentemente, de etanol. Sendo assim, o objetivo deste trabalho foi avaliar diferentes estratégias, a fim de obter melhoria na produção de etanol, a partir da fração de celulose presente na casca de arroz, utilizando a levedura termotolerante Kluyveromyces marxianus ATCC 36907. A primeira estratégia consistiu em realizar a SSF em batelada alimentada, objetivando o aumento da carga de sólidos; a segunda foi avaliar o efeito da adição de surfactantes não iônicos durante a hidrólise enzimática no processo de SSF. Primeiramente, a biomassa de casca de arroz foi submetida à caracterização química quanto aos teores de celulose, hemicelulose e lignina, antes e após pré-tratamento alcalino com hidróxido de sódio (NaOH) 8% (m/v). A casca de arroz tratada foi submetida à hidrólise enzimática com diferentes cargas enzimáticas (5, 11 e 22 FPU/gcelulose) e 10% de sólidos (m/v), a fim de determinar a condição com maior liberação de açúcares redutores para posterior utilização na SSF em batelada alimentada com alta carga de sólidos. Para a SSF em batelada alimentada, foram realizados quatro experimentos com diferentes concentrações de biomassa de cascas de arroz, sendo os experimentos E1 e E2, com carga inicial de 10% e final de 20% (m/v) e E3 e E4 com carga inicial de 15% e final de 25% (m/v), respectivamente, em pH 5,5, 40 °C, 200 rpm, 96 horas. Para isso, foram estabelecidos intervalos de alimentação, sendo duas alimentações a cada 24 horas (E1 e E3) e quatro alimentações a cada 12 horas (E2 e E4). Como segunda estratégia, foi realizada a hidrólise enzimática, com o objetivo de avaliar o efeito de diferentes cargas de enzima (5; 8; 13,5; 19 e 22 FPU/gcelulose), em associação aos surfactantes Polisorbato 80 (PS80) (7,0; 10; 15; 20 e 23 mg/L), Polietilenoglicol 4000 (PEG) (0,5; 1,0; 2,0; 3,0 e 3,5 g/L) e Triton X-100 (TX-100) (75; 100; 150; 200 e 225 mg/L), na liberação de glicose, por meio de três experimentos, seguindo um delineamento composto central rotacional (DCCR) 22, que inclui 4 ensaios nas condições axiais e 4 repetições no ponto central, totalizando 12 ensaios para cada surfactante, além de um experimento controle (com adição de enzima e sem surfactante), em triplicata. A terceira estratégia consistiu na adição de surfactante na SSF, objetivando avaliar a influência do surfactante na produção de etanol. Para isso, foram realizados três experimentos: controle (sem surfactante+19 FPU/gcelulose), com adição de Polisorbato 80 (10,0 mg/L+19 FPU/gcelulose) e com Polietilenoglicol 4000 (3,0 g/L+19 FPU/gcelulose), sendo o tipo de surfactante e respectivas concentrações e carga enzimática selecionados a partir dos resultados da vii hidrólise enzimática, que foram significativos. Os experimentos foram feitos com uma proporção de sólidos de 10% (m/v), pH 5,5, 40 °C, 200 rpm, 96 horas, em triplicata. A casca de arroz in natura apresentou 36,03% de celulose, 14,71% de hemicelulose e 26,47% lignina e, após o tratamento com NaOH 8% (m/v), os teores dessas frações foram 70,27; 8,39 e 21,34% de celulose, hemicelulose e lignina, respectivamente. Na SSF em batelada alimentada com alta carga de sólidos, obteve-se produção de etanol de 9,57; 9,92; 13,99 e 12,77 g/L, rendimentos (YE/C) de 0,16;0,17; 0,18 e 0,18 g/g e produtividade volumétrica (QP) de 0,27; 0,28; 0,39 e 0,53 g/L.h -1 , para os experimentos E1, E2, E3 e E4, respectivamente. Com relação ao efeito da adição de surfactantes na hidrólise enzimática, as concentrações de glicose variaram de 32,36 a 79,57 g/L, com adição de PS80, de 45,14 a 74,60 g/L, com PEG e 32,28 a 47,89 g/L, com adição de TX-100, enquanto o controle apresentou de 20,36 a 47,77 g/L, em 96 horas. Apenas os surfactantes PS80 e PEG, em associação a diferentes cargas de enzimas, promoveram efeitos significativos (p<0,05), sendo a carga enzimática o fator que mais influenciou o aumento da produção final de glicose. Em relação à SSF com surfactante, a adição de PS80 (10 mg/L) e PEG (3,0 g/L), em associação a 19 FPU/gcelulose, resultou em 6,73 e 8,17 g/L de etanol, YE/C de 0,17 e 0,21 g/g e QP de 0,56 e 0,68 g/L.h-1 , em 12 horas de fermentação, respectivamente. No controle, com adição da mesma carga enzimática (19 FPU/gcelulose) e sem surfactante, a produção de etanol foi 5,02 g/L, com YE/C de 0,13 g/g e QP de 0,14 g/L.h-1, em 36 horas. Os resultados sugerem que a casca de arroz é uma matéria-prima que pode ser utilizada como substrato em bioprocessos, por apresentar teor de celulose semelhante ao de outras biomassas lignocelulósicas. As diferentes estratégias de alimentação com casca de arroz, utilizadas na SSF em batelada alimentada, não resultaram aumentos substanciais na produção de etanol. A adição dos surfactantes PS80 e PEG, durante a hidrólise enzimática, resultou aumentos significativos à produção final de glicose. Na SSF, os mesmos surfactantes contribuíram para um ligeiro aumento na produção de etanol em um tempo menor de fermentação, comparado ao controle.Submitted by Neusa Fagundes (neusa.fagundes@unioeste.br) on 2020-10-09T18:37:45Z No. of bitstreams: 2 Lillian_Leonel2020.pdf: 4361117 bytes, checksum: 9cd9adff8da0bb3e0c2b696210dddf3a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2020-10-09T18:37:45Z (GMT). No. of bitstreams: 2 Lillian_Leonel2020.pdf: 4361117 bytes, checksum: 9cd9adff8da0bb3e0c2b696210dddf3a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2020-03-12Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfpor6588633818200016417500Universidade Estadual do Oeste do ParanáCascavelPrograma de Pós-Graduação em Engenharia AgrícolaUNIOESTEBrasilCentro de Ciências Exatas e Tecnológicashttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessBiomassa lignocelulósicaHidrólise enzimáticaBatelada alimentadaSurfactantesLignocellulosic biomassEnzymatic hydrolysisFed batchSurfactantsCIENCIAS AGRARIAS::ENGENHARIA AGRICOLAEstratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arrozEstrategies for the improvement of ethanol production by Kluyveromyces marxianus by simultaneous saccharification and fermentation process of rice husksinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-5347692450416052129600600600600221437444286838201591854457215887615552075167498588264571reponame:Biblioteca Digital de Teses e Dissertações do UNIOESTEinstname:Universidade Estadual do Oeste do Paraná (UNIOESTE)instacron:UNIOESTEORIGINALLillian_Leonel2020.pdfLillian_Leonel2020.pdfapplication/pdf4361117http://tede.unioeste.br:8080/tede/bitstream/tede/5020/5/Lillian_Leonel2020.pdf9cd9adff8da0bb3e0c2b696210dddf3aMD55CC-LICENSElicense_urllicense_urltext/plain; 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dc.title.por.fl_str_mv Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
dc.title.alternative.eng.fl_str_mv Estrategies for the improvement of ethanol production by Kluyveromyces marxianus by simultaneous saccharification and fermentation process of rice husks
title Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
spellingShingle Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
Leonel, Lillian Vieira
Biomassa lignocelulósica
Hidrólise enzimática
Batelada alimentada
Surfactantes
Lignocellulosic biomass
Enzymatic hydrolysis
Fed batch
Surfactants
CIENCIAS AGRARIAS::ENGENHARIA AGRICOLA
title_short Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
title_full Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
title_fullStr Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
title_full_unstemmed Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
title_sort Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz
author Leonel, Lillian Vieira
author_facet Leonel, Lillian Vieira
author_role author
dc.contributor.advisor1.fl_str_mv Sene, Luciane
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/2582084888410031
dc.contributor.referee1.fl_str_mv Arruda, Priscila Vaz de
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/1583339937667600
dc.contributor.referee2.fl_str_mv Cunha, Mário Antônio Alves da
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/3151576713472624
dc.contributor.referee3.fl_str_mv Kadowaki, Marina Kimiko
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/1819723253019762
dc.contributor.referee4.fl_str_mv Gomes, Simone Damasceno
dc.contributor.referee4Lattes.fl_str_mv http://lattes.cnpq.br/3362104483832351
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9257508293525181
dc.contributor.author.fl_str_mv Leonel, Lillian Vieira
contributor_str_mv Sene, Luciane
Arruda, Priscila Vaz de
Cunha, Mário Antônio Alves da
Kadowaki, Marina Kimiko
Gomes, Simone Damasceno
dc.subject.por.fl_str_mv Biomassa lignocelulósica
Hidrólise enzimática
Batelada alimentada
Surfactantes
topic Biomassa lignocelulósica
Hidrólise enzimática
Batelada alimentada
Surfactantes
Lignocellulosic biomass
Enzymatic hydrolysis
Fed batch
Surfactants
CIENCIAS AGRARIAS::ENGENHARIA AGRICOLA
dc.subject.eng.fl_str_mv Lignocellulosic biomass
Enzymatic hydrolysis
Fed batch
Surfactants
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::ENGENHARIA AGRICOLA
description Rice husk is a waste derived from rice milling and is an abundant byproduct in cereal producing countries. Due to the considerable contents of cellulose in its composition, it is a potential raw material for use in various bioprocesses, such as ethanol production. However, several technical challenges must be overcome to make the second-generation ethanol production process economically viable and competitive. In this context, the process of simultaneous saccharification and fermentation (SSF), associated with the increase of solids concentration, as well as the use of surfactants in enzymatic hydrolysis in fermentation processes that employ lignocellulosic biomass are promising strategies to obtain higher sugar yields and consequently ethanol. Thus, the objective of this work was to evaluate different strategies to obtain an improvement in ethanol production from rice husk cellulose, using the thermotolerant yeast Kluyveromyces marxianus ATCC 36907. The first strategy was to carry out the fed batch SSF to increase the solids load; the second was to evaluate the effect of adding nonionic surfactants during enzymatic hydrolysis in the SSF process. First, rice husk biomass was subjected to chemical characterization for cellulose, hemicellulose, and lignin content, before and after alkaline pretreatment with 8% (w/v) sodium hydroxide (NaOH). The pretreated rice husk was subjected to enzymatic hydrolysis with different enzyme loading (5, 11, 22 FPU/gcelullose) and 10% solids (w/v) to determine the condition with increased release of reducing sugars for subsequent SSF use in batch fed with high solids loading. For the SSF in fed batch, four experiments were performed with different biomass concentrations of rice husk, being E1 and E2 experiments with initial load of 10% and final 20% (w/v) and E3 and E4 with initial load 15% and final 25% (w/v), respectively, at pH 5.5; 40 °C, 200 rpm, 96 hours. Thus, feeding intervals were established, with two feedings every 24 hours (E1 and E3) and four feedings every 12 hours (E2 and E4). As a second strategy, enzymatic hydrolysis was carried out, with the objective of evaluating the effect of different enzyme loads (5; 8; 13.5; 19 and 22 FPU/gcellulose) it was evaluated in association with Polysorbate 80 (PS80) surfactants (7.0, 10, 15, 20 and 23 mg/L), Polyethylene glycol 4000 (PEG) (0.5, 1.0, 2.0, 3.0 and 3.5 g/L) and Triton X-100 (TX-100) (75; 100; 150, 200 and 225 mg/L) in glucose release through three experiments following a central rotational composite design (DCCR) 22, including 4 axial conditions and 4 center point repetitions, totaling 12 assays for each surfactant, besides a control experiment (with enzyme addition and without surfactant), in triplicate. The third strategy consisted of adding the surfactant SSF, to evaluate the influence of the surfactant in the production of ethanol. For this, three experiments were performed: control (without surfactant + 19 FPU/gcellulose), with addition of Polysorbate 80 (10.0 mg/L + 19 FPU/gcellulose) and with Polyethylene Glycol 4000 (3.0 g/L + 19 FPU/gcellulose), the type of surfactant and respective concentrations and enzyme load selected from the results of enzymatic hydrolysis that were significant. The experiments were carried out with a solids ratio of 10% (m / v), pH 5.5, 40 °C, 200 rpm, 96 hours, in triplicate. The chemical composition of rice husk was 36.03% cellulose, 14.71% hemicellulose and 26.47% lignin and after pretreatment with 8% NaOH (w/v) the content of these fractions was 70.27; 8.39 and 21.34% cellulose, hemicellulose, and lignin, respectively. In the fed batch SSF with high solids loading, an ethanol production was obtained of 9.57; 9.92; 13.99 and 12.77 g/L, yields (YE/C) of 0.16; 0.17; 0.18 and 0.18 g/g and volumetric productivity (QP) of 0.27; 0.28; 0.39 and 0.53 g/L.h-1 for the experiments E1, E2, E3 and E4, respectively. For the enzymatic hydrolysis, glucose concentrations ranged from 32.36 to 79.57 g/L with PS80 addition, from 45.14 to 74.60 g/L with PEG and 32.28 to 47.89 g/L with addition of TX-100, while the control showed from 20.36 to 47.77 g/L in 96 hours. Only the ix surfactants PS80 and PEG in association with different enzyme loads promoted significant effects (p<0.05), the enzymatic load being the factor that most influenced the increase of final glucose production. Regarding SSF with surfactant, the addition of PS80 (10 mg/L) and PEG (3.0 g/L) in association with 19 FPU/gcellulose resulted in 6.73 and 8.17 g/L of ethanol, YE/C of 0.17 and 0.21 g / g and QP of 0.56 and 0.68 g/Lh-1 , in 12 hours of fermentation, respectively. The results suggest that rice husk is a raw material that can be used as a substrate in bioprocesses because it has a cellulose content similar to that of other lignocellulosic biomasses. The different rice husk feeding strategies used in fed batch SSF did not result in substantial increases in ethanol production. The addition of surfactants PS80 and PEG during enzymatic hydrolysis resulted in significant increases in final glucose production. In SSF, the same surfactants contributed to a slight increase in ethanol production in a shorter fermentation time, compared to the control.
publishDate 2020
dc.date.accessioned.fl_str_mv 2020-10-09T18:37:45Z
dc.date.issued.fl_str_mv 2020-03-12
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.identifier.citation.fl_str_mv LEONEL, Lillian Vieira. Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz. 2020. 102 f. Tese (Programa de Pós-Graduação em Engenharia Agrícola) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.
dc.identifier.uri.fl_str_mv http://tede.unioeste.br/handle/tede/5020
identifier_str_mv LEONEL, Lillian Vieira. Estratégias visando à melhoria da produção de etanol por Kluyveromyces marxianus pelo processo de sacarificação simultânea à fermentação de cascas de arroz. 2020. 102 f. Tese (Programa de Pós-Graduação em Engenharia Agrícola) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.
url http://tede.unioeste.br/handle/tede/5020
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dc.publisher.none.fl_str_mv Universidade Estadual do Oeste do Paraná
Cascavel
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dc.publisher.initials.fl_str_mv UNIOESTE
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Centro de Ciências Exatas e Tecnológicas
publisher.none.fl_str_mv Universidade Estadual do Oeste do Paraná
Cascavel
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