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Doctoral Thesis
DOI
https://doi.org/10.11606/T.18.2019.tde-17062019-111032
Document
Author
Full name
Raissa Cristina da Silva Mazareli
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2019
Supervisor
Committee
Silva, Maria Bernadete Amancio Varesche (President)
Delforno, Tiago Palladino
Gomes, Simone Damasceno
Gorni, Guilherme Rossi
Sivieri, Katia
Title in Portuguese
Otimização da produção de hidrogênio a partir de resíduos de banana: avaliação da diversidade de bactérias autóctones e distribuição funcional
Keywords in Portuguese
bioenergia
enzimas
metagenômica
reator anaeróbio
resíduo frutícola
Abstract in Portuguese
A banana (Musa spp.) está entre as culturas mais abundantes no mundo, e devido ao seu descarte, desde a colheita até a sua comercialização gera-se grande quantidade de resíduos sólidos. Neste cenário, buscou-se avaliar o potencial uso do resíduo de banana (RB) como substrato e fonte de bactérias fermentativas para produção de H2 e metabólitos solúveis. Obteve-se a partir da fermentação natural do RB biomassa autóctone produtora de H2 sem necessidade do uso de fonte exógena de bactérias e custo adicional de fontes de carbono. Nesse consórcio foram identificadas bactérias semelhantes a Lactobacillus e Clostridium. Ensaios em reatores em batelada foram conduzidos para seleção do meio de cultivo (BAC, Noparati e PCS), das variáveis independentes (pH, temperatura, concentração do substrato, headspace e inóculo), seguido do delineamento composto central rotacional (DCCR) para otimização da produção de hidrogênio. Verificou-se que as condições nutricionais do meio PCS (extrato de levedura, peptona, NaCl e CaCO3) e RB resultaram na produção (P) e rendimento (YH2) máximo de 15,05 mL e 10,03 mL H2.g-1 CT (carboidratos totais), respectivamente. Em relação às variáveis independentes (pH, temperatura, concentração de substrato, volume do headspace e porcentagem de inóculo) obteve-se maiores valores de P e velocidade de produção (Rm) de H2 de 38,08 mL e 3,07 mL.h-1, respectivamente, em pH 7,5, 15 g.L-1, 44ºC, 40% de headspace e 15% inóculo. Em relação às variáveis estatisticamente significativas (pH e temperatura) via realização do DCCR observou-se que o aumento do pH (de 5,09 para 7,5) favoreceu, tanto P, quanto Rm, todavia, maior temperatura (de 27,1 para 46,9ºC) associada ao menor pH (<6,5) foram condições desfavoráveis para esses parâmetros. Por outro lado, obteve-se redução do tempo de início de produção de hidrogênio (λH2) para maior temperatura (44-46,9ºC) associada ao menor pH (5,5) As condições operacionais ótimas estimadas via modelo foram em pH 7,0 e 37ºC, obtendo-se 70,09 mL H2, 12,43 mL H2.h-1 e 93 mL.g-1 CT, para P, Rm e YH2, respectivamente. Bacilllus sp. isolada do RB cresceu em variedade de substratos (glicose, xilose, manose, galactose, frutose, maltose, celobiose, sacarose, amido e RB), e 71 mL H2 foi obtido em 5 g.L-1 de RB, pH 7 a 37ºC. Em todos os ensaios em reatores em batelada, independentemente das condições operacionais, verificou-se que as principais vias metabólicas foram do ácido acético butírico e ácido lático, principalmente com glicose e frutose como fonte de carbono. A acidificação dos reatores resultou na diminuição do pH inicial para valores <4,0 favorecendo a rota solventogênica. Na biomassa fermentativa autóctone e aquela dos reatores em batelada foi possível inferir sobre elevado potencial metabólico, devido a identificação de genes relacionados com enzimas do metabolismo de carboidratos.
Title in English
Optimization of hydrogen production from banana residue: evaluation of autochthonous bacteria diversity and functional distribution
Keywords in English
anaerobic reactor
bioenergy
enzymes
fruit waste
metagenomics
Abstract in English
Banana (Musa spp.) is among the most abundant crops in the world and, due to its discard, great quantity of solid waste is generated from the harvest until its commercialization. In this scenario, the potential use of banana residue (RB) as substrate and source of fermentative bacteria for the production of H2 and soluble metabolites was evaluated. It was obtained from RB natural fermentation native autochthonous H2-producing biomass without the need to use exogenous source of bacteria and additional costs from carbon sources. In this consortium, bacteria similar to Lactobacillus and Clostridium were identified. Batch reactor experiments were conducted to select culture medium (BAC, Noparati and PCS) and independent variables (pH, temperature, substrate concentration, headspace and inoculum), followed by central composite rotatable design (CCRD) experiments for optimization of hydrogen production. It was verified that the nutritional conditions provided by PCS medium (yeast extract, peptone, NaCl and CaCO3) and RB resulted in the maximum production (P) and yield (YH2) of 15.05 mL and 10.03 mL H2 g-1 CT (total carbohydrates), respectively. Regarding the independent variables (pH, temperature, substrate concentration, headspace volume and percentage of inoculum), higher P and H2 production rate (Rm) were obtained (38.08 mL and 3.07 mL h-1, respectively) at pH 7.5, 15 g L-1 of substrate, 44 °C, 40% headspace and 15% inoculum. In relation to the statistically significant variables (pH and temperature), it was observed through CCRD that the increase in pH (from 5.09 to 7.5) favored both P and Rm, however, a higher temperature (from 27.1 to 46.9ºC) associated with lower pH (<6.5) were unfavorable conditions for these parameters. On the other hand, a reduction in the hydrogen production start time (H2) was obtained with higher temperature (44-46.9ºC) associated to lower pH (5.5). The optimum operational conditions, estimated by modeling, were pH 7.0 and 37 °C, yielding 70.09 mL H2, 12.43 mL H2 h-1 and 93 mL gCT-1 for P, Rm and YH2, respectively. Bacillus sp. isolated from RB grown on a variety of substrates (glucose, xylose, mannose, galactose, fructose, maltose, cellobiose, sucrose, starch and RB), and 71 mL H2 was obtained with 5 g L-1 RB, pH 7 at 37 °C. In all batch reactor experiments, regardless of operating conditions, the main metabolic pathways were acetic acid, butyric acid and lactic acid, mainly with glucose and fructose as carbon sources. Acidification of the reactors resulted in a decrease of the initial pH to values <4.0, favoring the solventogenic pathway. It was possible to infer about the high metabolic potential in the autochthonous fermentative biomass and in the biomass from the batch reactors due to the identification of genes related to enzymes of the carbohydrate metabolism.
 
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Publishing Date
2019-07-01
 
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