Orientador: Maria Helena Andrade Santana / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-20T01:59:45Z (GMT). No. of bitstreams: 1 Benedini_LeandroJunqueira_M.pdf: 2379873 bytes, checksum: 6e878845e2b38c395cbc006441849b4c (MD5) Previous issue date: 2012 / Resumo: O ácido hialurônico (AH) é um polissacarídeo linear com vasta gama de aplicações médicas e farmacêuticas. Por esta razão, é essencial a obtenção do produto com elevado grau de pureza. Atualmente, há grande foco de pesquisa direcionado ao AH produzido por rota microbiana, utilizando extrato de leveduras (EL) como fonte de nitrogênio. Disso resulta uma quantidade relevante de proteínas e peptídeos totais (TPP) remanescentes no meio de cultivo. Este trabalho visa estudar o efeito da substituição do EL e de fontes de origem animal, por peptonas vegetais no cultivo do Streptococcus zooepidemicus para a produção do AH, com finalidades de reduzir a quantidade de TPP nos meios e eliminar possíveis contaminantes. Na primeira etapa deste trabalho, foram realizadas fermentações com peptonas de trigo (PT), de batata (PB), de soja (PS) e uma mistura de peptonas vegetais (MPV). O extrato de leveduras (EL) foi usado como controle. Os melhores resultados foram obtidos em meio suplementado com PS e com PT. Meio composto com a primeira peptona (PS) produziu 0,28 g.'L POT. -1'de AH com 1,4 g de TPP por g de AH, enquanto meio suplementado com EL apresentou 0,21 g.'L POT. -1' de AH com 4,1 g de TPP por g de AH. Meio composto com a segunda peptona (PT) produziu menor quantidade de AH (0,05 g.'L POT. -1'), porém com maior massa molar (5,06.106 Da) que a observada em AH proveniente de outras fontes de nitrogênio (3,2 - 3,7 x '10 POT. 6' Da). Numa segunda etapa, foram realizadas novas fermentações com finalidade de se realizar um estudo cinético do metabolismo microbiano com as peptonas mais promissoras da etapa anterior (PS e PT). Novamente, meio suplementado com PS apresentou o melhor desempenho para a produção. Enquanto isso, PT apresentou menor desempenho para produção de AH, mas AH com maior massa molar média (4,76 x '10 POT. 6' Da). Além disso, análises cinéticas demonstraram que quanto maior quantidade de AH produzido, menor a sua massa molar. Estes resultados reforçam os benefícios de utilização PS para produção de grandes quantidades de AH, e os benefícios da utilização de PT para produção de AH com elevado peso molecular médio. Numa terceira etapa, foi estudado o efeito da substituição dos meios BHI (Brain and Heart Infusion) e sangue de carneiro por PS no preparo de placas utilizadas para adaptação e crescimento celular em etapa inicial da inoculação. Quando somente BHI foi substituído por PS, e a presença de sangue de carneiro foi mantida, quantidade semelhante de AH foi produzida em relação ao controle. A remoção de sangue de carneiro das placas com a melhor concentração de de quantidade semelhante de AH com maior massa molar média (3,50 - 3,60 x '10 POT. 6' Da). Portanto, foi concluído que PS e PT substituem EL em meio de fermentação, produzindo maior quantidade de AH, e AH com maior massa molar, respectivamente. Além disso, PS se demonstrou efetiva para crescimento e desenvolvimento de células em placas, já que substitui BHI e sangue de carneiro / Abstract: Hyaluronic acid (HA) is a linear polysaccharide with a wide range of medical and pharmaceutical applications. For this reason, it is essential to obtain a product with high purity. Currently, HA is produced by microbial route, using yeast extract (YE) as nitrogen source. This results in a significant amount of total proteins and peptides (TPP) remaining in the culture medium. It was studied effects of replacing YE by vegetable peptones in the cultivation of Streptococcus zooepidemicus for HA production, with purposes of reducing the amount of TPP in the media and to remove potential contaminants. In the first step of this work, fermentations were performed during 24 hours in 100 ml of medium with wheat (WP), potato (PP) and soy (SP) peptones and a mixture of vegetable peptones (MPV). The yeast extract (YE) was used as control. The best results were obtained in medium supplemented with SP and WP. The first peptone (SP) produced 0.30 g.'L POT. -1' of HA with 1.4 g of TPP per g of HA, while medium supplemented with YE produced 0.21 g.'L POT. -1' of HA with 4.1 g of TPP per g of HA. The second peptone (WP) produced less amount of HA (0.05 g.'L POT. -1'), but with higher molecular weight (5.06 x '10 POT. 6' Da) than observed in HA from other sources of nitrogen (3.2 - 3.7 x '10 POT. 6' Da). In a second step, fermentation was carried out with 340 ml of medium, during 33 hours, with the purpose of conducting a microbial kinetics study with the most promising peptones of the previous step (SP and WP). Again, SP showed the best performance for production. Meanwhile, WP showed lower performance for HA production, but higher HA average molar mass. These results underscore the benefits of using SP for the production of large amounts of HA and WP utilization for production of HA with high average molecular weight. In a third step, it was studied the effect of replacing the BHI media ("Brain and Heart Infusion") and sheep blood by SP in the preparation of Petri plates used for adaptation and cell growth in the initial stage of inoculation. When only BHI was replaced by 67 g.'L POT. -1' SP, and the presence of sheep blood was maintained, 0.27 g.'L POT. -1' of HA was produced in the fermentation (24 hours, 100 ml medium), against 0.29 g.'L POT. -1' using 37 g. 'L POT. -1' of BHI and sheep blood. Removal of sheep blood from Petri plates with the best concentration of SP (67 g.'L POT. -1') led to the production of 0.30 g.'L POT. -1' HA. Furthermore, HA in fermentation from SP plates performed a higher average molar mass (from 3.50 to 3.60 x '10 POT. 6' Da) compared with HA obtained from plates with sheep blood and BHI (3.09 x '10 POT. 6' Da ). These results also demonstrated effectiveness of SP on the growth of Streptococcus zooepidemicus on plates. It was concluded that SP and WP replace YE in fermentation medium, producing higher amount of HA and HA with higher molecular weight, respectively. Furthermore, SP has been shown effective for cell growth and development / Mestrado / Desenvolvimento de Processos Biotecnologicos / Mestre em Engenharia Química

The bacterium Clostridium acetobutylicum produces acetone and n [subscript] - butanol from molasses in an industrial fermentation system. Although the bacterium has been cultured in liquid media it does not grow well on agar plates and requires high concentrations of hydrogen. Pretreatment of agar plates with bovine catalase improves growth on agar media. The bacteria produce an area of clearing (halo) on Potato agar plates due to butyric acid (the precursor of n [subscript]-butanol) and ß -amylase production. This characteristic will be used as a plate screening assay for the selection of high solvent producing mutants. A laboratory scale fermentation system was developed and detailed studies including pH, turbidity and cell morphology changes, and the details of solvent production were undertaken. The fermentation was optimized for mutant selection. The production of normal solvent yields by isolated clones is required for the mutant selection programme. Studies revealed that sporulation of the clones increased their solvent yield although solvent yields were still lower than normal. Efficient sporulation is therefore a prerequisite for clone fermentation. The origin of the phage infection during the factory outbreak was determined and resistant clones obtained. The presence of a bacteriocin-like toxin causing decreases in turbidity was identified during the final fermentation stage. The strain sensitivity, optimum conditions for stability as well as the kinetics of inactivation and lethality have been investigated. Preliminary characterization and purification studies indicate the proteinaceous nature of the toxin. / KMBT_363

Feasibility of the biodegradation of neutralized-caustic potato peel waste by aerobic-thermophilic fermentation was examined. Fermentation temperature, pH, rate of solids utilization, dissolved oxygen and total organic nitrogen were characterized with respect to the fermentation process. Examination of these parameters showed distinct characteristic behaviour, though the parameter magnitude varied between the trials. The naturally occurring flora of thermophilic microorganisms of the potato was sufficient as an inoculum, with their propagation creating the fermentation characteristics. The solids content of the potato waste was reduced by the thermophilic fermentation with a noticeable improvement in the nitrogen content in the remaining residue. Protein and amino acid analyses supported the feasibility of using the remaining residue as animal feed. Increases in the nitrogen content of the substrate was assumed to be due to the conversion of atmospheric nitrogen by nitrogen-fixing microorganisms within the fermentation. Microbial identification indicated that these organisms belonged to the species Bacillus coagulans. No strains within this species have been previously reported to fix atmospheric nitrogen. It was concluded that an aerobic-thermophilic fermentation can be utilized as a waste treatment process for caustic potato peel waste. Besides the numerous advantages of an aerobic-thermophilic fermentation, the validity for the utilization of this fermentation process in the treatment of potato peel waste is the near total disposal of the waste. / Land and Food Systems, Faculty of / Graduate

M.Tech. (Chemical Engineering) / Please refer to full text to view abstract

Orientadores: Ariovaldo José da Silva, Bruna de Souza Moraes / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Agrícola / Made available in DSpace on 2018-08-30T07:46:17Z (GMT). No. of bitstreams: 1 Santos_GracieteMarydos_M.pdf: 3271887 bytes, checksum: 539f68130eb11eac441d2bca07c2e7da (MD5) Previous issue date: 2015 / Resumo: No Brasil, o efluente industrial produzido em maior quantidade é a vinhaça, caracterizada por altos níveis de ácidos orgânicos, fósforo, cálcio, potássio e magnésio. O reaproveitamento energético da vinhaça mostra-se como uma alternativa interessante para produção de biocombustíveis ou sub-produtos. Este trabalho avaliou o potencial da vinhaça como fonte de substrato e nutrientes para produção de álcoois e ácidos orgânicos voláteis (AOV) por meio fermentação em batelada utilizando consórcio anaeróbio (lodo de bovinocultura) pré-tratados com choque térmico (TT) e choque ácido-térmico (AT). Foram utilizados dois meios diferentes, de sacarose (S) e de vinhaça (V), sendo a sacarose a principal fonte de carbono. A vinhaça provou ser uma excelente fonte de nutrientes para os microrganismos envolvidos na fermentação butírica, uma vez que a adição de vinhaça melhora significativamente a produção de ácido butírico em comparação com meio de cultura sintético. As máximas concentrações de ácido butírico, iso-butírico e acético foram de 14,13 ± 0,77 g L-1 na amostra ATV B3; 10,34 ± 0,43 g L-1na amostra ATV B2 e; 4,13 ± 0,06 g L-1na amostra TTV B3, respectivamente. O rendimento dos AOV acético, iso-butírico e butírico e de etanol foi mais elevado nas amostras ATV B3 e TTV B3, atingindo valores máximos de 0,14; 0,28; 0,69 e; 0,26 g g-1 carboidratos totais, respectivamente. Não foram encontradas diferenças significativas entre métodos de pré-tratamento e enriquecimento de inóculo, AT e TT no que diz respeito a produção de ácido butírico e etanol. Em escala maior, operando em reator de 1,5 L, a fermentação de vinhaça bruta e melado de cana por consórcio microbiano AU mostrou potencial para produção de solventes como o butanol, uma vez que concentrações elevadas de ácido butírico foram produzidas, com concentração máxima, rendimento e produtividade de 13,85 g L-1; 0,64 g g-1 e; 199,98 mg L h-1, respectivamente. A caracterização microbiológica, pirosequenciamento, revelou a ocorrência em maior abundância de bactérias do gênero Clostridium, principalmente no consórcio AU e Lactobacillus mais abundante nos consórcios TT e AT. Foi identificada uma espécie conhecida pela produção de butanol, o C. pasteurianum no consórcio AU. Contudo, o presente trabalho representa um passo importante no desenvolvimento de um processo industrial para reutilização da vinhaça. A exploração de novos microrganismos e estudo dos fatores que interferem no processo de fermentação como pH, temperatura, nutrientes, densidade da cultura, cargas aplicadas e características do substrato, são fundamentais para o entendimento dos efeitos sinérgicos e antagônicos da associação de culturas / Abstract: In Brazil, industrial waste produced in the greatest amount is vinasse, characterized by high levels of organic acids, phosphorus, calcium, potassium and magnesium. The energy reuse of vinasse shows up as an interesting alternative for the production of biofuels or byproducts. This study evaluated the potential of vinasse as a source of substrate and nutrients for the production of alcohols and volatile fatty acids (VFA) through fermentation batch using anaerobic consortium (cattle sludge) pre-treated with heat shock (TT) and acid-shock thermal (AT). We used two different media, sucrose (S) and vinasse (V), with sucrose being the main source of carbon. The vinasse proved to be an excellent source of nutrients for microorganisms involved in the butyric fermentation, since the addition of vinasse significantly improves the production of butyric acid as compared to synthetic culture medium. The maximum concentrations of butyric acid, iso-butyric and acetic acid were 14.13 ± 0.77 g L-1 in the sample ATV B3; 10.34 ± 0.43 g L-1 in ATV B2 and 4.13 ± 0.06 g L-1 in TTV B3, respectively. The yield of acetate, iso-butyric acid, butyrate and ethanol was higher in ATV B3 and TTV B3 samples, reaching maximum values of 0.14; 0.28; And 0.69; 0.26 g g-1 total carbohydrates, respectively. There were no significant differences between pretreatment and enrichment methods inoculum, TA and TT as regards the production of butyric acid and ethanol. On a larger scale, operating at 1.5 L reactor, crude fermentation vinasse and molasses of sugar cane from AU microbial consortium showed potential for producing butanol as the solvent, since high concentrations of butyric acid was produced, with maximum concentration, yield and productivity of 13.85 g L-1 0.64 g g-1 and 199.98 mg h L-1, respectively. Microbiological characterization, pyrosequencing, revealed the occurrence in greater abundance of the genus Clostridium bacteria, particularly the AU and most abundant Lactobacillus in consortium TT and AT consortia. C. pasteurianum, known for the production of butanol was identified in AU consortium. However, this study represents an important step in the development of an industrial process for reuse of vinasse. The exploration of new microorganisms and study of the factors that interfere in fermentation process such as pH, temperature, nutrients, cultures, applied loads and characteristics of the substrate are critical for understanding the synergistic and antagonistic effects of culture associatio / Mestrado / Agua e Solo / Mestra em Engenharia Agrícola

Two series of batch and fed-batch fermentations were carried out using S.cerevisiae in a semi-defined medium containing 200 gl-1 glucose as limiting substrate. Growth rates were calculated and the data used to test the applicability of eight empirical kinetic models. The form proposed by Levenspiel, combining the concept of a limiting ethanol concentration with a power-law form, gave the best results with these data. Glucose concentration was found to have a far smaller, though not negligible, effect on growth rate under these conditions. It was also observed that in fed-batch fermentations the total substrate uptake rate of the broth became constant soon after commencement of feeding, without cessation of growth. It is suggested that ethanol inhibits the synthesis of a rate-controlling enzyme in the glycolyti·c chain, but no previous work could be found to support or refute this explanation. A quasi-mechanistic model of growth under the condition of constant substrate consumption rate is formulated and discussed.

No description available.

The purpose of this study is to investigate the rate of natural fermentation of solutions at different temperatures. Whatever microorganisms that chanced to be present in the air and that chanced to fall into the inoculating medium are the ones which brought about fermentation when transferred to the fermentable solution.

Ethanol produced from lignocellulosic agricultural products and waste is an environmentallyriendly alternative to petroleum-derived fuel. Lignocellulosic biomass is gasified producing synthesis gas, which is composed of CO, CO2, and H2. Synthesis gas is fermented via anaerobic biocatalyst. The bacterium was grown in a fructose-rich medium then concentrated in ethanol production medium for synthesis gas fermentation. While the known ethanol-producing bacterium Clostridium ljungdahlii was used to provide baseline values for synthesis gas utilization and ethanol production, synthesis gas fermentation were conduced with a culture discovered at Mississippi State University. Additionally, efforts were made to isolate other anaerobic cultures capable of fermenting synthesis gas to ethanol.

No description available.