Made available in DSpace on 2017-05-12T14:47:55Z (GMT). No. of bitstreams: 1 Andrea Christina Larsen.pdf: 426110 bytes, checksum: 0afb74a50b694490128340543e09daa9 (MD5) Previous issue date: 2009-07-23 / The National Biodiesel Production and its Application Program, created by the Federal Government, introduces biodiesel in the Brazilian energetic matrix, bringing along the perspective to improve glycerin offer, a co-product of biodiesel production. Since for each 100 L of biodiesel, 10 kg of raw glycerin are produced, this implies on the development of studies for an economically viable use of glycerin. So, the main objective of this paper was to evaluate the addition of raw glycerin in anaerobic digestion of cassava starch production wastewater, in a semi-continuous-flux reactor of one-phase in bench scale. A 8.77 L reactor, with useful volume, was used, with PVC conduit as a support medium, temperature of 26±1º C, supplied with cassava starch production wastewater and glycerin, with hydraulic detention times of 4 and 5 d and organic loads of 3.05, 9.32, 14.83 e 13.59 g COD L-1 d-1, in this order, obtained with the addiction of glycerin at 0, 2, 3 and 2%, respectively. The biogas production per reactor volume increased with the 2% glycerin addiction with regard to the control treatment. However, in the next treatments, biogas production decreased when organic load increased. The major average biogas production per reactor volume was 1.979 L biogas L-1 d-1, for the organic load of 9.32 g COD L-1 d-1. The major averages of biogas production per g COD and g TVS consumed were 0.397 and 0.661 L, for the control treatment, without glycerin addition. The best COD removal, 98.69%, occurred for 2% glycerin treatment and 9.32 g COD L-1 d-1 organic load, followed by 96.41% for the control treatment. The major removal of TS and TVS occurred for the control treatment, for 2% glycerin treatment and organic load of 9.32 g COD L-1 d-1, by 81.19 and 75.47% TS and 90.21 and 81.18% TVS for the respective treatments. Thus, the evaluated reactor in this trial can be used for the treatment of cassava starch production wastewater with crude glycerin addition as a way of giving a correct destination to the last one, without using neutralizers and nutrients. / O Programa Nacional de Produção e Uso do Biodiesel, lançado pelo Governo Federal, introduz o biodiesel na matriz energética brasileira com a perspectiva de aumento da oferta de glicerina, co-produto gerado na produção do biodiesel. Uma vez que, para cada 100 L de biodiesel são produzidos 10 kg de glicerina bruta, isto implica o desenvolvimento de estudos para o aproveitamento economicamente viável da glicerina. O objetivo principal deste trabalho foi avaliar a adição de glicerina bruta na digestão anaeróbia de efluente de fecularia, em reator de fluxo semi-contínuo horizontal de uma fase, em escala de laboratório. Foi utilizado reator com volume útil de 8,77 L, com meio suporte em conduítes de PVC, temperatura de 26±1º C, alimentado com manipueira e glicerina, com tempos de detenção hidráulica de 4 e 5 d e cargas orgânicas de 3,05, 9,32, 14,83 e 13,59 g DQO L-1 d-1, nesta ordem, obtidas com a adição de glicerina aos níveis de 0, 2, 3 e 2%, respectivamente. A produção de biogás por volume de reator aumentou com a adição de 2% de glicerina em relação ao tratamento controle. Porém, nos tratamentos seguintes, a produção de biogás diminuiu com o aumento da carga orgânica. A maior produção média de biogás por volume de reator foi de 1,979 L biogás L-1 d-1, para a carga de 9,32 g DQO L-1 d-1. As maiores produções médias de biogás por g DQO e g STV consumidos foram de 0,397 e 0,661 L, para o tratamento controle, sem adição de glicerina. A maior remoção de DQO, 98,69%, ocorreu para o tratamento com 2% de glicerina e carga de 9,32 g DQO L-1 d-1, seguida de 96,41% para o tratamento controle. As maiores remoções de ST e STV ocorreram para o tratamento controle, para o tratamento com 2% de glicerina e carga de 9,32 g DQO L-1 d-1, na ordem de 81,19 e 75,47% de ST e 90,21 e 81,18% de STV para os respectivos tratamentos. Assim, o reator avaliado neste trabalho pode ser utilizado para o tratamento de efluente de fecularia com adição de glicerina bruta como forma de dar destinação correta a essa última, sem a utilização de neutralizantes e nutrientes.

O Banco de Tecidos do Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo está desenvolvendo uma linha de estudos experimentais com o intuito de apresentar novos métodos de armazenamento de aloenxertos. Este estudo tem como objetivo comparar o método da criopreservação (- 80° C) com o da conservação em glicerol a 98% em temperatura ambiente. Foram analisadas tanto a capacidade de inibição de crescimento bacteriano e fúngico quanto às eventuais alterações histológicas. Para isso, foram estudadas 30 amostras de tecido ósseo trabecular coletadas de 10 pacientes submetidos à artroplastia total do quadril. Estas amostras foram divididas em três grupos de 10 espécimes, a saber: grupo controle, grupo criopreservado e grupo conservado em glicerol a 98% à temperatura ambiente. O período de armazenamento das amostras foi de um ano. Os resultados foram analisados estatisticamente pelo método de McNemar, com índice de significância de 0,10. No que diz respeito à preservação da matriz óssea, não houve variações significativas nos dois grupos estudados em relação ao grupo controle. Não ocorreu crescimento bacteriano ou de fungos nas amostras armazenadas durante um ano em glicerol a 98%. Por ser extremamente reduzido em relação à criopreservação, o custo do método da conservação em glicerol a 98% o indica para uso em Bancos de Tecidos de pequena monta. Contudo, são ainda necessários posteriores estudos sobre as propriedades biomecânicas, remoção do glicerol do tecido ósseo e o processo de integração biológica dos mesmos com o receptor. / The Tissue Bank of Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Universidade de São Paulo is developing a line of experimental studies with the intention to present new methods of allografts storage. This study has the objective to compare the method of cryopreservation (-80 ºC) with glycerol 98% preservation at room temperature. It even analyses, in such way, the inhibition capacity of bacterial and fungal growth and any eventual histological changes. For this, 30 samples of trabecular bone tissue have been collected from 10 patients submitted to total hip arthroplasty. These samples were separated in three groups of 10 specimens, as following: a control group, a cryopreserved group and a glycerol 98% preserved group at room temperature. They were stored during a period of one year. The results were analysed by the McNemar statistic method, with a significance of 0,10. Concerning to bone matrix, no significant changes occurred to the two studied groups compared to the control group. Bacterial and fungal growth do not occurred in the stored samples for one year in glycerol 98%. For being extremely reduced when compared with cryopreservation method, the preservation cost in glycerol 98% indicates its use on small sum tissue banks. However, posterior studies about biomechanical properties, glycerol removal of bone tissue and their biological process of integration with the receiver are necessary.

Tendo em vista a busca por matérias-primas de fontes renováveis e menos poluentes, e o crescente aproveitamento de subprodutos gerados por estas fontes, surgiu o interesse em estudar a síntese do carbonato de glicerina a partir da glicerina obtida como subproduto principal na fabricação de biodiesel. Sintetizou-se este composto no Laboratório de Química Orgânica Aplicada. Este tensoativo não é disponível no Brasil e possui propriedades cosméticas interessantes, que, no entanto, ainda devem ser mais exploradas na atualidade. No estudo, adicionou-se o carbonato a sabões líquidos obtidos a partir dos ácidos graxos provenientes dos óleos de soja e de mamona. Os sabões produzidos apresentaram baixa alcalinidade, reduzindo, assim, as adições de agentes químicos que podem ser agressivos à pele, como mostraram os resultados parciais. A partir do planejamento experimental, verifica-se a influência da temperatura e o tempo de reação da síntese do carbonato, assim como o tipo de catalisador utilizado. Análises de espectroscopia vibracional Raman e infravermelho, juntamente com ensaios de viscosidade, mostraram que a temperatura ideal para a carboxilação é de 140 °C, sendo que o tempo reacional deve ser de 6 a 7 h, utilizando-se como catalisador o sulfato de magnésio. Demonstrou-se também, a sensibilidade das duas técnicas na identificação dos picos das reações de síntese de carbonato de glicerila. Comparativamente, foram realizados testes de espuma dos sabões adicionados de carbonato de glicerila e de lauril sulfato de sódio. De um modo geral, a adição de carbonato de glicerila aos sabões propiciou a formação de uma espuma mais densa e persistente. A utilização de água dura reduziu o volume de espuma formado para todas as amostras. Comparando os dois óleos vegetais, soja e mamona, percebeu-se que os sabões produzidos a partir de óleo de mamona apresentaram menor formação de espuma. A adição de dietanolamina reduziu significativamente a estabilidade das espumas em todas as amostras. Estudos reológicos demonstraram que o tensoativo carbonato de glicerila é um fluido newtoniano, comportamento semelhante ao da glicerina pura. / In order to search for raw materials from cleaner and renewable sources, and the growing use of byproducts generated by these sources, has appeared the interest in studying the synthesis of glycerol carbonate from glycerol obtained as a main byproduct in the manufacture of biodiesel. This compound was synthesized at the Laboratory of Applied Organic Chemistry. This surfactant is not available in Brazil and has interesting cosmetic properties, which, however, should be more exploited at present. In the study, carbonate was added to liquid soaps from the fatty acids from soybean oil and castor oil. Soaps produced had low alkalinity, thereby reducing the additions of chemicals that can be aggressive to the skin, as partial results showed. From the experimental design, the influence of temperature and reaction time of the synthesis of carbonate is investigated, as well as the type of catalyst used. Analysis of partial Raman vibrational spectroscopy, together with viscosity experiments showed that the optimal temperature for carboxylation is 140 °C, and the reaction time should be from 6 to 7 h, using magnesium sulphate as the catalyst. It also demonstrated the sensitivity of both techniques in identifying the peaks of the reactions of synthesis of glycerol carbonate. In comparison,foam tests were made of soaps added glycerol carbonate and sodium laurylsulfate. Overall, the addition of glycerol carbonate to soap provided the formation of denser and persistent foam. The use of hard water reduced the volume of foam formed for all samples. Comparing the two vegetable oils, soy and castor bean, it was noticed that soaps produced from castor oil had lower foaming. The addition of diethanolamine significantly reduced the stability of foams in all samples. Rheological studies demonstrated that the surfactant glycerol carbonate is neuwtonian fluid, behavior similar to that of pure glycerin.

Systemic acquired resistance (SAR) is a type of plant defense mechanism that is induced after a localized infection and confers broad-spectrum immunity against related or unrelated pathogens. During SAR, a number of chemical signals and proteins generated at the site of primary infection travel to the uninfected tissues and are thought to alert the distal sites against secondary infections. Glycerol-3-phosphate (G3P) is one of the chemical signals that play an important role in SAR. G3P is synthesized in the cytosol and chloroplasts via the enzymatic activities of G3P Dehydrogenase (G3Pdh) or Glycerol Kinase (GK). Interestingly, a mutation in three of the five G3Pdh isoforms or GK impairs SAR by lowering the pathogen induced G3P pool. This suggests that total cellular pool of G3P is critical for SAR. To determine factors contributing to G3P flux between various subcellular compartments I analyzed the role of putative G3P transporters in G3P flux and SAR. The Arabidopsis genome encodes five isoforms of G3P Permeases (G3Pp) and these transmembrane proteins are predicted to localize to plasma membrane, chloroplast or mitochondria. At least two G3Pp isoforms (G3Pp1 and G3Pp3) were able to complement the Escherichia coli mutant impaired in the uptake of G3P into the cytoplasm. Characterization of Arabidopsis G3Pp mutants showed that a mutation in G3Pp2, G3Pp3 and G3Pp4 compromised SAR but not local resistance. Furthermore, this SAR defect could only be complemented by exogenous application of G3P. The G3Pp mutants accumulated wild-type-like levels of G3P suggesting that the subcellular compartmentalization of G3P might contribute to the induction of SAR.

Based on previous studies of the novel Lactobacillus reuteri DPC16 strain, an in vitro investigation on the supernatant antimicrobial activity and the culture safety against normal gastrointestinal microflora and gastric mucus was done in this thesis. DPC16 cell-free supernatants (fresh and freeze-dried, designated as MRSc and FZMRSc) from anaerobic incubations in pre-reduced MRS broth, have shown significant inhibitory effects against selected pathogens, including Salmonella Typhimurium, E. coli O157:H7, Staphylococcus aureus, and Listeria monocytogenes. These effects were mainly due to the acid production during incubation as evidenced by the negation of such activity from their pH-neutral counterparts, and this acidic effect was shown to reduce the pathogen growth rate and decrease the total number of pathogen cells. By incubation of concentrated (11 g/L) resting cells in glycerol-supplemented MRS broth, another DPC16 cell-free supernatant (designated as MRSg) has shown very strong antimicrobial effect against all target pathogens. As indicated by a kinetic profile, this activity developed in a sigmoidal fashion as incubation proceeded, reaching to maximum activity after 6-8h and maintained at the same level thereafter. Further study has shown that the antimicrobial activity of this supernatant was pH-independent, effective across a pH range of 4.6 to 6.5, and acted on both Gram-negative and Gram-positive pathogens. Using the minimum effective dose, a time course investigation has provided evidence that this supernatant affected the growth of the target pathogens by elongating the lag phase and lowering the total cell number at the end of the incubation. Lastly, it was found that the strong antimicrobial effect of MRSg was bactericidal at high concentrations and bacteriostatic at low concentrations. However, it also found that the viability of DPC16 cells also decreased as incubation prolonged, which suggests that this glycerol-derived supernatant had a lethal effect to its own cells. Nevertheless, this lethal effect was exerted to a much lesser extent compared with that to the pathogens. The last DPC16 cell-free supernatant was designated as SGF, which was produced from secondary fermentation of the same resting cells in glycerol-water. SGF did not show a significant antimicrobial activity, which suggests that this specific strain is not capable of utilising glycerol in the absence of fermentable carbohydrates. The antimicrobial activity found in MRSg matched with previously published characteristics of reuterin, which is a unique antimicrobial substance synthesised by L. reuteri when incubated with glycerol. Therefore, a study on the production kinetics of reuterin by DPC16 was carried out. Supernatants of both MRSg and SGF were studied. Results showed that glycerol utilisation occurred in both MRSg and SGF, whereas the bioconversion of glycerol into reuterin was different. In MRSg, glycerol was constantly utilised by DPC16 resting cells, and by the end of an 18h incubation 85.8 mM of glycerol was utilised, where 72.8% was transformed into reuterin. The formation of reuterin initiated with an inclining production and reached the maximum rate of 10.9 mM/h after 6h of incubation, with the total production of 64 mM of reuterin at the end of the 18h incubation. This reuterin production in MRSg followed a similar pattern to that of its antimicrobial activity, which suggests a certain correlation between reuterin formation and the increase of antimicrobial activity in MRSg. Therefore, the major antimicrobial component in MRSg that accounted for its potent antimicrobial activity was very much likely to be reuterin. In SGF, however, detectable reuterin was negligible even though some glycerol may have been absorbed into the highly concentrated DPC16 resting cells. This has responded to the antimicrobial activity assay in that due to the lack of essential carbohydrate nutrient for normal cell metabolism, there was no glycerol utilisation and hence no reuterin synthesis. Having studied the antimicrobial activity of L. reuteri DPC16 and the production of antimicrobial-competent reuterin, two safety issues (the impact on growth on other normal commensal probiotics and mucin degradation activity) of this strain were assessed to further evaluate its probiotic potential. By using similar in vitro assays as in the antimicrobial test, the same set of DPC16 supernatants have demonstrated no adverse effect on the growth of either Lactobacillus acidophilus, Lactobacillus plantarum, Pediococcus acidilactici, or Bifidobacterium lactis DR10. No stimulatory effect was found either. By incorporating purified porcine gastric mucin into classic mucin-degradation assays in both liquid and agar media, DPC16 has not exhibited the same mucinolytic activity as that of the faecal flora cultures. Thus, it can be concluded that L. reuteri DPC16 is as safe to the host as normal commensal microflora.

Acrolein is an indispensable chemical intermediate with a rising demand in recent years. The concern of the increase of propylene prices due to the shrinking supply of nonrenewable crude oil makes the acid-catalyzed gas-phase glycerol dehydration to acrolein a prime candidate for research. Our analysis showed that the sustainable acrolein production from glycerol was both technically and economically viable. Alumina2700® (Al) and Silica1252® (Si) loaded with silicotungstic acid (HSiW) possessed distinct features while provided equally good acrolein yield (73.86mol% and 74.05mol%, respectively) optimally. Due to the unique non-equilibrium characteristics, non-thermal plasma (NTP) could promote a variety of chemical reactions; however, its application in a dehydration process remained blank. This study used the reaction of glycerol dehydration to acrolein to probe whether NTP could 1) improve acrolein yield during dehydration, 2) suppress the coke formation and regenerate the catalyst, and 3) modify the properties of the catalyst. The dielectric barrier discharge configuration was used to generate NTP; various NTP field strengths and also their interaction with temperature and the catalyst were investigated. The results showed that NTP improved the glycerol conversion and that NTP with a proper field strength increased acrolein selectivity. The optimal acrolein yields of 83.6 mol% and 83.1 mol% were achieved with 3.78 kV/cm NTP and 4.58 kV/cm NTP at 275°C for HSiW-Al and HSiW-Si, respectively. The application of NTP-O2 (5% oxygen in argon, 4.58 kV/cm) during glycerol dehydration significantly suppressed coke formation on HSiW-Si. NTP-O2 could regenerate the deactivated HSiW-Si at low temperatures by removing both soft and hard coke at various rates. NTP-O2 with higher field strength, at medium operation temperature (150ºC) and in argon atmosphere was more effective for coke removal/catalyst regeneration. Applying NTP to the catalyst fabrication showed some capabilities in modifying catalyst properties, including enlarging surface area, preserving mesopores, increasing acid strength and Brønsted acidity. NTP with argon as the discharge gas performed better in these modifications than NTP with air as the discharge gas.

This thesis presents the mathematical modeling of two new Escherichia coli platforms with economical potential for the production of biofuels and chemicals, namely glycerol fermentation and the reversal of the β-oxidation cycle. With the increase in traditional fuel prices, alternative renewable energy sources are needed, and the efficient production of biofuels becomes imperative. So far studies have focused on using glucose as feedstock for the production of ethanol and other fuels, but a recent increase in glycerol availability and its consequent decrease in price make it an attractive feedstock. Furthermore, the reversed β-oxidation cycle is a highly efficient mechanism for the synthesis of long-chain products. These two platforms have been reported experimentally in E. coli but their mathematical modeling is presented for the first time here. Because mathematical models have proved to be useful in the optimization of microbial metabolism, two complementary models were used in this study: kinetic and stoichiometric. Kinetic models can identify the control structure within a specific pathway, but they require highly detailed information, making them applicable to small sets of reactions. In contrast, stoichiometric models require only mass balance information, making them suitable for genome-scale modeling to study the effect of adding or removing reactions for the optimization of the synthesis of desired products. To study glycerol fermentation, a kinetic model was implemented, allowing prediction of the limiting enzymes of this process: glycerol dehydrogenase and di-hydroxyacetone kinase. This prediction was experimentally validated by increasing their enzymatic activities, resulting in a two-fold increase in the rate of ethanol production. Additionally, a stoichiometric genome-scale model (GEM) was modified to represent the fermentative metabolism of glycerol, identifying key metabolic pathways for glycerol fermentation (including a new glycerol dissimilation pathway). The GEM was used to identify genetic modifications that would increase the synthesis of desired products, such as succinate and butanol. Finally, glucose metabolism using the reversal β-oxidation cycle was modeled using a GEM to simulate the synthesis of a variety of medium and long chain products (including advanced biofuels). The model was used to design strategies that can lead to increase the productivity of target products.

The main objective of this thesis was to study the anaerobic digestion (AD) of wastewater from a chicken processing facility and of crude glycerol from local biodiesel operations. The AD of these substrates was conducted in bench-scale reactors operated in the batch mode at 35°C. The secondary objective was to evaluate two sources of glycerol as co-substrates for AD to determine if different processing methods for the glycerol had an effect on CH₄ production. The biogas yields were higher for co-digestion than for digestion of wastewater alone, with average yields at 1 atmosphere and 0°C of 0.555 and 0.540 L (g VS added)⁻¹, respectively. Another set of results showed that the glycerol from an on-farm biodiesel operation had a CH₄ yield of 0.702 L (g VS added)⁻¹, and the glycerol from an industrial/commercial biodiesel operation had a CH₄ yield of 0.375 L (g VS added)⁻¹. Therefore, the farm glycerol likely had more carbon content than industrial glycerol. It was believed that the farm glycerol had more impurities, such as free fatty acids, biodiesel and methanol. In conclusion, anaerobic co-digestion of chicken processing wastewater and crude glycerol was successfully applied to produce biogas rich in CH₄.

The study was to investigate the changes in (1) triacylglycerol (TAG) contents and its relationship to (2) lipid synthesis- and metabolism-related enzyme activity and (3) their gene expression in cobia (Rachycentron canadum) during the fast growth period (from October 2006 to April 2007) in ventral muscle and liver in Hsiao-Lu-Chiao island in southwestern Taiwan. The crude lipid was 12% for fed diet, 30-40% for liver while 13% in February and 11% to 9% in other month for muscle. The TAG content of crude lipid was 36 % for fed diets, and from 22% (December) to 40% (February) for muscle, and from 63% (October to February) to 47% (March) for liver. Oil red-O (ORO) staining showed that TAG accumulated in muscle in February but in December in liver. Muscle TAG contents and enzyme activities and mRNA levels of GPDH and FAS increased in February. A decrease in GPDH enzyme activity and mRNA levels but an increase in PEPCK enzyme activity and mRNA levels indicate the increased supply of acetyl-CoA for fatty acid synthesis is in muscle. An increase in FATP2 mRNA levels suggest the influx of fatty acid also contributes to increased fatty acid accumulation in muscle.In liver, TAG and fatty acid contents decreased in March April but increased FAS and PEPCK enzyme activity and mRNA levels. It is possible that fatty acid synthesis is enhanced in March, but a fast transport to other organs results in a net decline in liver fatty acid contents and subsequently a decrease in TAG contents. FATP contents decreased in March-April mRNA, indicating that the influx of fatty acid in decreasing in liver in adult fish. GPDH and GAPDH were not related to lipid metabolism in liver. These data from enzyme activity and mRNA level, demonstrated that a potentially increase in acetyl-CoA via PEPCK contributes to fatty acid synthesis and GPDH-mediated synthesis of G-3-P provide the C skeleton for TAG synthesis.

The increased use of fossil fuels during the last centuries has caused elevated levels of carbon dioxide in the atmosphere. There is significant evidence that this is the cause of global warming. To mitigate the global warming, measures has to be taken to use renewable fuels and make processes more efficient. Catalytic gasification and downstream upgrading of synthesis gas is a promising technology for biofuel production, where previous research in black liquor gasification is currently expanding into a wider fuel feedstock. This work focuses on co-gasification of black liquor and by-products from other biofuel production technologies. The interesting by-products were crude glycerol from biodiesel production and spruce fermentation residue from ethanol production. The main goals were to study if the fuels can mix homogeneously and study the char reactivity. CO2 char gasification for mixtures of black liquor and glycerol or fermentation residue respectively was studied using thermogravimetric analysis (TGA) for four temperatures between 750°C and 900°C. The results show that glycerol can be mixed in all proportions with black liquor and indicate that the char reactivity is unchanged. The sustained char reactivity for blends is attributed to the volatility of glycerol. The fermentation residue does not produce a homogeneous mixture with black liquor and the char is less reactive. More studies should be performed to further elucidate the validity of the results.