Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/29839 |
Resumo: | The rice husk (RH) conversion in products with industrial and pharmaco-chemical attention aimed to use this raw lignocellulosic material, renewable and abundant in the south of Brazil, mainly at RS state ─ yet with no commercial value. The main goal was the production of exopolysaccharides (EPS) and, secondarily, to contribute to mitigate the improper disposal of this waste of processing. For this, the RH was submitted to alkaline pretreatment (KOH, Ca(OH)2, NaOH, CaCO3 and NH4OH) and/or by ozonation, followed by acidic pressurized (between 10 – 40 bar) hydrolysis with H2SO4, HCl, HNO3 and H3PO4, according to multivariate statistical design, using a central rotational compound design. The hydrolytical step was evaluated in three different configurations: a system consisting in a heating block (Berghof), a normal lab stove and a lab microwave oven. The hydrolytic experiments in heating block showed the best result at 135 °C and 55 min of reaction, with 4.9% (v v -1) H3PO4, producing 20.9 g L-1 of sugar (yield of 209.4 mg g-1 RH). Employing 2.5% (v v -1) H2SO4, 145 °C and 45 min of reaction, it was produced 16.1 g L-1 of sugar, yield of 161.5 mg g-1 RH in the best experiment with lab stove. Using a microwave oven, the better experiment yielded 15.3 g L-1 of sugar (yield 152.6 mg g-1 RH), using 1.5% (v v-1) HCl 135 °C and 55 min of reaction. Regarding the results, it is possible to distinguish the action profile of each acid over the RH: H2SO4 or HCl mainly release glucose as HNO3 or H3PO4 generate better xylose yields. The best results of the alkaline hydrolysis were 21.7 g L-1, yield increase of 51% by using KOH (65 ° C, 60 min, alkali at 4.25% m v-1), 24.0 g L-1, yield increase of 66% with NaOH (65 ° C, 35 min, alkali at 4.25% m v-1). The use of ozonation, on the other hand, besides its character residue-free, resulted in productivities of 16.1 g L-1 (yield rise of 11%). Previous fermentative experiments for the growth of the bacterium Rhizobium radiobacter (LMG196 and ATCC4720) in the RH hydrolysate were successful. A multivariate design evaluated the culture conditions (10% inoculums volume (1x109 UFC mL-1), pH 7.0, 100 rpm, 72 h) and yeast extract supplementation (9.0 g L-1), KH2PO4 (1.0 g L-1) and FeSO4. Under these conditions it is possible to produce extractable EPS (by using cooled isopropanol) with yields of 69.0 g L-1 (41.6 g 100 g-1 CA), significant value from 72 h culture growth (with sulfuric hydrolysate detoxified with 4.0% m v-1 active carbon). The analytical techniques used were liquid chromatography, HPLC-RID and HPLC-DAD, and UV-vis spectrophotometry, discussing the figures-of-merit and validation methodologies developed. For the product characterization it was used the analytical techniques of NMR 1H, FT-IR and GPC. It was demonstrated the viability of the fermentable sugar production process from the RH, on a laboratory scale and, also, the feasibility of its bioconversion in EPS (purity of 91.2%), which contributes markedly to the effort of the development of biotechnology research and scientific innovation. |
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Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arrozBiotechnological production of exopolysaccharide by chemical-fermentative conversion of rice huskCasca de arrozHidrólise ácida à pressãoRhizobium radiobacterExopolissacarídeosOtimização multivariada de processosRice husksPressurized acid hydrolysisExopolysaccharidesMultivariate optimization processCNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICAThe rice husk (RH) conversion in products with industrial and pharmaco-chemical attention aimed to use this raw lignocellulosic material, renewable and abundant in the south of Brazil, mainly at RS state ─ yet with no commercial value. The main goal was the production of exopolysaccharides (EPS) and, secondarily, to contribute to mitigate the improper disposal of this waste of processing. For this, the RH was submitted to alkaline pretreatment (KOH, Ca(OH)2, NaOH, CaCO3 and NH4OH) and/or by ozonation, followed by acidic pressurized (between 10 – 40 bar) hydrolysis with H2SO4, HCl, HNO3 and H3PO4, according to multivariate statistical design, using a central rotational compound design. The hydrolytical step was evaluated in three different configurations: a system consisting in a heating block (Berghof), a normal lab stove and a lab microwave oven. The hydrolytic experiments in heating block showed the best result at 135 °C and 55 min of reaction, with 4.9% (v v -1) H3PO4, producing 20.9 g L-1 of sugar (yield of 209.4 mg g-1 RH). Employing 2.5% (v v -1) H2SO4, 145 °C and 45 min of reaction, it was produced 16.1 g L-1 of sugar, yield of 161.5 mg g-1 RH in the best experiment with lab stove. Using a microwave oven, the better experiment yielded 15.3 g L-1 of sugar (yield 152.6 mg g-1 RH), using 1.5% (v v-1) HCl 135 °C and 55 min of reaction. Regarding the results, it is possible to distinguish the action profile of each acid over the RH: H2SO4 or HCl mainly release glucose as HNO3 or H3PO4 generate better xylose yields. The best results of the alkaline hydrolysis were 21.7 g L-1, yield increase of 51% by using KOH (65 ° C, 60 min, alkali at 4.25% m v-1), 24.0 g L-1, yield increase of 66% with NaOH (65 ° C, 35 min, alkali at 4.25% m v-1). The use of ozonation, on the other hand, besides its character residue-free, resulted in productivities of 16.1 g L-1 (yield rise of 11%). Previous fermentative experiments for the growth of the bacterium Rhizobium radiobacter (LMG196 and ATCC4720) in the RH hydrolysate were successful. A multivariate design evaluated the culture conditions (10% inoculums volume (1x109 UFC mL-1), pH 7.0, 100 rpm, 72 h) and yeast extract supplementation (9.0 g L-1), KH2PO4 (1.0 g L-1) and FeSO4. Under these conditions it is possible to produce extractable EPS (by using cooled isopropanol) with yields of 69.0 g L-1 (41.6 g 100 g-1 CA), significant value from 72 h culture growth (with sulfuric hydrolysate detoxified with 4.0% m v-1 active carbon). The analytical techniques used were liquid chromatography, HPLC-RID and HPLC-DAD, and UV-vis spectrophotometry, discussing the figures-of-merit and validation methodologies developed. For the product characterization it was used the analytical techniques of NMR 1H, FT-IR and GPC. It was demonstrated the viability of the fermentable sugar production process from the RH, on a laboratory scale and, also, the feasibility of its bioconversion in EPS (purity of 91.2%), which contributes markedly to the effort of the development of biotechnology research and scientific innovation.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqA conversão da casca de arroz (CA) em produtos de interesse industrial e farmoquímico objetivou o aproveitamento desta matéria-prima lignocelulósica, renovável e abundante na região sul brasileira, ainda sem valor comercial. A principal meta foi a produção de exopolissacarídeo (EPS) e, secundariamente, contribuir para mitigar a questão da disposição imprópria deste resíduo de beneficiamento do arroz. Para tanto, a CA foi submetida à pré-tratamento alcalino (KOH, Ca(OH)2, NaOH, CaCO3 e NH4OH) e/ou por ozonização, seguido de hidrólise ácida à pressão (entre 10-40 bar) com H2SO4, HCl, HNO3 e H3PO4, segundo planejamentos estatísticos multivariados. A etapa hidrolítica foi avaliada em três diferentes configurações: bloco de aquecimento (Berghof), estufa e forno micro-ondas. Os experimentos hidrolíticos, em bloco de aquecimento, apresentaram o melhor resultado a 135 °C e 55 min de reação, com 4,9% (v v-1) H3PO4, tendo sido produzidos 20,9 g L-1 de açúcar (rendimento de 209,4 mg g-1 de CA). Empregando-se estufa, o experimento com 2,5% (v v-1) H2SO4, 145 ºC e 45 min de reação, produziu 16,1 g L-1 de açúcar, com rendimento de 161,5 mg g-1 CA. Utilizando-se forno micro-ondas, o melhor experimento produziu 15,3 g L-1 de açúcar (rendimento de 152,6 mg g-1 CA), empregando-se 1,5 % (v v-1) HCl, 135 ºC e 55 min de reação. Com base nos resultados, distingue-se o perfil de ação de cada ácido sobre a CA: H2SO4 e HCl liberam majoritariamente glicose, enquanto HNO3 e H3PO4 geram melhores rendimentos em xilose. Os melhores resultados da etapa alcalina resultaram em 22,7 g L-1, acréscimo de rendimento de 58% com emprego de KOH (65 °C, 60 min, 6,5% m v-1 do álcali), 24,0 g L-1, acréscimo de 67% com NaOH (80 °C, 35 min, 6,5% m v-1 do álcali). O emprego de ozonização, por outro lado, além de ser livre de resíduos, resultou em produtividades de 16,1 g L-1 (ganho de 11%). Ensaios fermentativos prévios atestaram o crescimento de Rhizobium radiobacter (LMG196 e ATCC4720) no hidrolisado de CA. Com abordagem multivariada sobre as condições de cultivo (10% de inóculo (1x109 UFC mL-1), pH 7,0, 100 rpm, 72 h), avaliou-se a suplementação com extrato de levedura (até 9,0 g L-1), KH2PO4 (1,0 g L-1) e FeSO4. Nestas condições ocorre a produção de EPS (succinoglucanas) passíveis de extração (com isopropanol resfriado), rendendo até 69 g L-1 (41,6 g 100 g-1 CA) de EPS, valor significativo obtido em 72 h de cultivo do hidrolisado (destoxificado com carvão ativo, 4% m v-1). As técnicas analíticas utilizadas foram cromatografia líquida, HPLC-RID e HPLC-DAD, e espectrofotometria UV-vis, apresentando-se figuras-de-mérito e validação das metodologias desenvolvidas. Para a caracterização do produto foram utilizadas as técnicas de RMN 1H, FT-IR, MEV e GPC. Foi demonstrada a viabilidade do processo de produção de açúcares fermentescíveis a partir da CA, em escala de laboratório e, igualmente, a viabilidade de sua bioconversão em EPS (91,2% de pureza), contribuindo acentuadamente para o esforço do desenvolvimento da pesquisa biotecnológica e da inovação científica.Universidade Federal de Santa MariaBrasilAdministraçãoUFSMPrograma de Pós-Graduação em AdministraçãoCentro de Ciências Naturais e ExatasMartins, Ayrton Figueiredohttp://lattes.cnpq.br/2113532494494821Mortari, SérgioMontipó, SheilaPaniz, José Neri GottfriedBolzan, RodrigoPedroso, Giovanni Bressiani2023-08-03T16:06:45Z2023-08-03T16:06:45Z2018-07-13info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/29839porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2023-08-03T16:06:45Zoai:repositorio.ufsm.br:1/29839Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2023-08-03T16:06:45Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz Biotechnological production of exopolysaccharide by chemical-fermentative conversion of rice husk |
title |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
spellingShingle |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz Pedroso, Giovanni Bressiani Casca de arroz Hidrólise ácida à pressão Rhizobium radiobacter Exopolissacarídeos Otimização multivariada de processos Rice husks Pressurized acid hydrolysis Exopolysaccharides Multivariate optimization process CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA |
title_short |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
title_full |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
title_fullStr |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
title_full_unstemmed |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
title_sort |
Produção biotecnológica de exopolissacarídeo por meio de conversão químico-fermentativa de casca de arroz |
author |
Pedroso, Giovanni Bressiani |
author_facet |
Pedroso, Giovanni Bressiani |
author_role |
author |
dc.contributor.none.fl_str_mv |
Martins, Ayrton Figueiredo http://lattes.cnpq.br/2113532494494821 Mortari, Sérgio Montipó, Sheila Paniz, José Neri Gottfried Bolzan, Rodrigo |
dc.contributor.author.fl_str_mv |
Pedroso, Giovanni Bressiani |
dc.subject.por.fl_str_mv |
Casca de arroz Hidrólise ácida à pressão Rhizobium radiobacter Exopolissacarídeos Otimização multivariada de processos Rice husks Pressurized acid hydrolysis Exopolysaccharides Multivariate optimization process CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA |
topic |
Casca de arroz Hidrólise ácida à pressão Rhizobium radiobacter Exopolissacarídeos Otimização multivariada de processos Rice husks Pressurized acid hydrolysis Exopolysaccharides Multivariate optimization process CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA |
description |
The rice husk (RH) conversion in products with industrial and pharmaco-chemical attention aimed to use this raw lignocellulosic material, renewable and abundant in the south of Brazil, mainly at RS state ─ yet with no commercial value. The main goal was the production of exopolysaccharides (EPS) and, secondarily, to contribute to mitigate the improper disposal of this waste of processing. For this, the RH was submitted to alkaline pretreatment (KOH, Ca(OH)2, NaOH, CaCO3 and NH4OH) and/or by ozonation, followed by acidic pressurized (between 10 – 40 bar) hydrolysis with H2SO4, HCl, HNO3 and H3PO4, according to multivariate statistical design, using a central rotational compound design. The hydrolytical step was evaluated in three different configurations: a system consisting in a heating block (Berghof), a normal lab stove and a lab microwave oven. The hydrolytic experiments in heating block showed the best result at 135 °C and 55 min of reaction, with 4.9% (v v -1) H3PO4, producing 20.9 g L-1 of sugar (yield of 209.4 mg g-1 RH). Employing 2.5% (v v -1) H2SO4, 145 °C and 45 min of reaction, it was produced 16.1 g L-1 of sugar, yield of 161.5 mg g-1 RH in the best experiment with lab stove. Using a microwave oven, the better experiment yielded 15.3 g L-1 of sugar (yield 152.6 mg g-1 RH), using 1.5% (v v-1) HCl 135 °C and 55 min of reaction. Regarding the results, it is possible to distinguish the action profile of each acid over the RH: H2SO4 or HCl mainly release glucose as HNO3 or H3PO4 generate better xylose yields. The best results of the alkaline hydrolysis were 21.7 g L-1, yield increase of 51% by using KOH (65 ° C, 60 min, alkali at 4.25% m v-1), 24.0 g L-1, yield increase of 66% with NaOH (65 ° C, 35 min, alkali at 4.25% m v-1). The use of ozonation, on the other hand, besides its character residue-free, resulted in productivities of 16.1 g L-1 (yield rise of 11%). Previous fermentative experiments for the growth of the bacterium Rhizobium radiobacter (LMG196 and ATCC4720) in the RH hydrolysate were successful. A multivariate design evaluated the culture conditions (10% inoculums volume (1x109 UFC mL-1), pH 7.0, 100 rpm, 72 h) and yeast extract supplementation (9.0 g L-1), KH2PO4 (1.0 g L-1) and FeSO4. Under these conditions it is possible to produce extractable EPS (by using cooled isopropanol) with yields of 69.0 g L-1 (41.6 g 100 g-1 CA), significant value from 72 h culture growth (with sulfuric hydrolysate detoxified with 4.0% m v-1 active carbon). The analytical techniques used were liquid chromatography, HPLC-RID and HPLC-DAD, and UV-vis spectrophotometry, discussing the figures-of-merit and validation methodologies developed. For the product characterization it was used the analytical techniques of NMR 1H, FT-IR and GPC. It was demonstrated the viability of the fermentable sugar production process from the RH, on a laboratory scale and, also, the feasibility of its bioconversion in EPS (purity of 91.2%), which contributes markedly to the effort of the development of biotechnology research and scientific innovation. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-07-13 2023-08-03T16:06:45Z 2023-08-03T16:06:45Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/29839 |
url |
http://repositorio.ufsm.br/handle/1/29839 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Administração UFSM Programa de Pós-Graduação em Administração Centro de Ciências Naturais e Exatas |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Administração UFSM Programa de Pós-Graduação em Administração Centro de Ciências Naturais e Exatas |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
_version_ |
1805922147868082176 |