Dissertação(mestrado)- Universidade Federal do Rio Grande, Programa de Pós-Graduação em Engenharia e Ciência de Alimentos, Escola de Química e Alimentos, 2009. / Submitted by Caroline Silva (krol_bilhar@hotmail.com) on 2012-08-23T22:10:06Z No. of bitstreams: 1 elisane-dissertao.pdf: 1678353 bytes, checksum: 827dc6807ea87ca51e1133f6f1dd934b (MD5) / Approved for entry into archive by Bruna Vieira(bruninha_vieira@ibest.com.br) on 2012-09-09T17:36:09Z (GMT) No. of bitstreams: 1 elisane-dissertao.pdf: 1678353 bytes, checksum: 827dc6807ea87ca51e1133f6f1dd934b (MD5) / Made available in DSpace on 2012-09-09T17:36:09Z (GMT). No. of bitstreams: 1 elisane-dissertao.pdf: 1678353 bytes, checksum: 827dc6807ea87ca51e1133f6f1dd934b (MD5) Previous issue date: 2009 / O glicerol gerado na síntese de biodiesel pode ser usado na produção de biomassa, para suplementação na alimentação, agregando valor a este subproduto. Este trabalho teve como objetivos: selecionar uma cepa de levedura promissora para produção de biomassa como fonte de proteínas, comparando a utilização de glicerol comercial puro e glicerol bruto proveniente da síntese de biodiesel, como principal fonte de carbono; selecionar o meio de cultivo, através do teste de Tukey; avaliar a composição do meio de cultivo de forma a maximizar a concentração de biomassa e conteúdo proteico através da técnica de planejamento experimental. Foram utilizadas as leveduras Yarrowia lipolytica NRRL YB- 423, Candida lipolytica NRRL Y-1095, Candida utilis NRRL Y-900 e Candida rugosa NRRL Y-95, certificadas como GRAS (Generally Recognized As Safe). Os cultivos foram realizados em frascos agitados, conduzidos em incubadora rotatória a 30°C e 180 rpm. As leveduras Yarrowia lipolytica YB-423 e Candida lipolytica Y-1095 apresentaram maior produção de proteína total em meio contendo glicerol puro, cerca de 1,9 e 1,7 g/L, respectivamente. Quando utilizado meio contendo glicerol bruto, proveniente da produção de biodiesel, não houve diferença estatisticamente significativa quanto à produção de proteína total entre as leveduras, mas quanto ao conteúdo proteico a levedura Candida utilis Y-900 apresentou diferença em relação às demais cepas. Quando avaliada a biomassa máxima, a levedura Yarrowia lipolytica YB-423 apresentou melhor resultado(14,7 g/L). Desta forma, a levedura Yarrowia lipolytica YB-423 foi selecionada para produção de biomassa como fonte de proteínas. Com esta levedura, foram estudados quatro meios de cultivo com composição distinta, sendo selecionado aquele com 50 g/L de glicerol, 5,5 g/L de hidrogenofosfato de amônio, 5,5 g/L de dihidrogenofosfato de potássio, 1 g/L de sulfato de amônio, 0,25 g/L de sulfato de magnésio, 0,021 g/L de cloreto de cálcio dihidratado, 1 g/L de extrato de levedura, 1 g/L de peptona, ajustado a pH 5,5, obtendo-se 18,2 % de conteúdo proteico, 17,8 g/L de biomassa máxima e 3,1 g/L de produção de proteína total. Um planejamento experimental fracionário 2v 5-1 foi utilizado para determinar as variáveis que mais influenciaram na produção de biomassa e no conteúdo proteico. Os parâmetros estudados foram pH inicial do meio de cultivo e as concentrações de glicerol, hidrogenofosfato de amônio, peptona e extrato de levedura. Posteriormente, as variáveis que mais influenciaram foram avaliadas em um planejamento experimental completo 23, fixando-se as concentrações de hidrogenofosfato de amônio (5,5 g/L) e peptona (1,5 g/L). Através da técnica de superfície de resposta, a condição para maximização do conteúdo proteico validada pelo modelo empírico foi de 33,9 g/L de glicerol, pH inicial de 4,8 e 0,6 g/L de extrato de levedura, obtendo-se um conteúdo proteico de 20,1± 0,6%, com uma biomassa máxima de 19,5 ± 1,0 g/L. / The glycerol generated in the biodiesel synthesis can be used in the production of biomass, for diet supplementation, adds value to this by-product. This work aims to: select a promising strain of yeast for production of biomass as a source of protein, comparing the use of pure commercial glycerol and crude glycerol from biodiesel synthesis as the main carbon source; select the culture media through the Tukey test; evaluate the composition of the culture medium in order to maximize the concentration of biomass and protein content by the experimental design technique. The yeast Yarrowia lipolytica YB-423, Candida lipolytica NRRL Y-1095, Candida utilis Y-900 and Candida rugosa Y-95, certified as GRAS (Generally Recognized As Safe) were used. The cultures were performed in shaken flasks, conducted in rotary incubator at 30°C and 180 rpm. Yarrowia lipolytica YB- 423 and Candida lipolytica Y-1095 showed higher production of total protein in the medium containing pure glycerol, about 1.9 and 1.7 g/L, respectively. In medium containing crude glycerol derived from the production of biodiesel, the yeasts showed no statistically significant difference in the production of total protein, but the protein content of the yeast Candida utilis Y-900 showed a difference in relation to the other strains. When assessing the maximum biomass, the yeast Yarrowia lipolytica YB-423 showed the best result (14.7 g/L). Therefore, the yeast Yarrowia lipolytica YB-423 was selected for production of biomass as a source of protein. With this yeast, we studied four culture media with different compositions, thereby selecting that with 50 g/L of glycerol, 5.5 g/L of ammonium hydrogen phosphate, 5.5 g/L of potassium dihydrogen phosphate, 1 g/L of ammonium sulphate, 0.25 g/L magnesium sulphate, 0.021 g/L dihydrated calcium chloride, 1 g/L yeast extract, 1 g/L peptone, pH adjusted to 5.5, resulting in 18.2% of protein content, 17.8 g/L of maximum biomass and 3.1 g/L of total protein production. A 2v 5-1 fractional experimental design was used to determine the variables that most influenced the production of biomass and protein content. The parameters studied were initial pH of the culture medium, glycerol concentrations, ammonium hydrogen phosphate, peptone and yeast extract. Subsequently, the variables that most influenced were evaluated in a full experimental design 23, setting up the concentrations of ammonium hydrogen phosphate (5.5 g/L) and peptone (1,5 g/L). Through the technique of surface response, the condition for maximizing the protein content validated by empirical model was 33.9 g/L glycerol, initial pH of 4.8 and 0.6 g/L yeast extract, resulting in a protein content of 20.1 ± 0.6%, with a maximum biomass of 19.5 ± 1.0 g/L.

Para a melhor extração da fração hemicelulósica do bagaço de cana-de-açúcar e obtenção do hidrolisado contendo uma mistura de açúcares, predominantemente de xilose, foi definido o processo de hidrólise com ácido na concentração de 100 mg H2S04/g m.s. a 140 °C por 20 minutos de reação. O hidrolisado hemicelulósico obtido foi utilizado para avaliação do crescimento celular e consumo de açúcares por 19 cepas de leveduras e 03 de fungos. Candida tropicalis lZ 1824 foi selecionada como promissora para produção de biomassa microbiana (11,0 g.l-1) e consumo de ART (89,8%) e apresentar fator de conversão (YK/S) de 0.50 g.g-1. Foi verificado ainda que a levedura selecionada apresentou melhores rendimentos em biomassa e consumo de ART, quando cultivada em hidrolisado suplementado com P2O5 (2.0 g.l-1) e uréia (2,0 g.l-1) a pH inicial 6,0. Finalmente verificou-se que na vazão específica de aeração de 2,0 v/v/m houve a maior velocidade específica máxima de crescimento (0,108 h-1), quando comparada com cultivos realizados a 1,0 v/v/m. A biomassa microbiana obtida nestas condições apresentou um teor protéico de 31, 3% com composição de aminoácidos comparável ao padrão da FAO. / For a better extraction of the hemicellulosic fraction from the sugar cane bagasse and obtention of hydrolysate containing a mixture of sugars, predominantly xylose, the hydrolysis process was defined, with acid in the concentration of 100 mg H2SO4/g dry matter, at 140 °C for 20 minutes of reaction. The hemicellulosic hydrolysate obtained was utilized for evaluation of cell growth and sugars consumption by 19 yeast stocks and 3 fungi stocks. Candida tropicalis lZ 1824 was selected for being considered as very promissing for production of microbial biomass (11,8 g.l-1) and TRS consumption (89,8%) and also for presenting a conversion factor (YK/S) of 0,50 g.g-1. It was noticed that the selected yeast gave greater biomass yields and TRS consumption when cultivated in hydrolysate supplemented with P2O5 (2,0 g.l-1) and urea (2,0 g.l-1) at initial pH 6,0. At last it was observed that the specific aeration flow of 2,0 v/v/m caused the highest maximum specific growth velocity (0,108 h-1) when compared to the cultivation accomplished of 1,0 v/v/m. The microbial bíomass obtained under these conditions presented a 31,3% protein content with a composition of amino acids comparable to the FAD standard.

Para a melhor extração da fração hemicelulósica do bagaço de cana-de-açúcar e obtenção do hidrolisado contendo uma mistura de açúcares, predominantemente de xilose, foi definido o processo de hidrólise com ácido na concentração de 100 mg H2S04/g m.s. a 140 °C por 20 minutos de reação. O hidrolisado hemicelulósico obtido foi utilizado para avaliação do crescimento celular e consumo de açúcares por 19 cepas de leveduras e 03 de fungos. Candida tropicalis lZ 1824 foi selecionada como promissora para produção de biomassa microbiana (11,0 g.l-1) e consumo de ART (89,8%) e apresentar fator de conversão (YK/S) de 0.50 g.g-1. Foi verificado ainda que a levedura selecionada apresentou melhores rendimentos em biomassa e consumo de ART, quando cultivada em hidrolisado suplementado com P2O5 (2.0 g.l-1) e uréia (2,0 g.l-1) a pH inicial 6,0. Finalmente verificou-se que na vazão específica de aeração de 2,0 v/v/m houve a maior velocidade específica máxima de crescimento (0,108 h-1), quando comparada com cultivos realizados a 1,0 v/v/m. A biomassa microbiana obtida nestas condições apresentou um teor protéico de 31, 3% com composição de aminoácidos comparável ao padrão da FAO. / For a better extraction of the hemicellulosic fraction from the sugar cane bagasse and obtention of hydrolysate containing a mixture of sugars, predominantly xylose, the hydrolysis process was defined, with acid in the concentration of 100 mg H2SO4/g dry matter, at 140 °C for 20 minutes of reaction. The hemicellulosic hydrolysate obtained was utilized for evaluation of cell growth and sugars consumption by 19 yeast stocks and 3 fungi stocks. Candida tropicalis lZ 1824 was selected for being considered as very promissing for production of microbial biomass (11,8 g.l-1) and TRS consumption (89,8%) and also for presenting a conversion factor (YK/S) of 0,50 g.g-1. It was noticed that the selected yeast gave greater biomass yields and TRS consumption when cultivated in hydrolysate supplemented with P2O5 (2,0 g.l-1) and urea (2,0 g.l-1) at initial pH 6,0. At last it was observed that the specific aeration flow of 2,0 v/v/m caused the highest maximum specific growth velocity (0,108 h-1) when compared to the cultivation accomplished of 1,0 v/v/m. The microbial bíomass obtained under these conditions presented a 31,3% protein content with a composition of amino acids comparable to the FAD standard.

Submitted by Maria Medeiros (maria.dilva1@ufcg.edu.br) on 2018-08-23T12:14:13Z No. of bitstreams: 1 MARCOS HENRIQUE SILVA ARAGÃO - DISSERTAÇÃO (PPGECA) 2017.pdf: 4540927 bytes, checksum: d25472de237d384e285a54b061998c1e (MD5) / Made available in DSpace on 2018-08-23T12:14:13Z (GMT). No. of bitstreams: 1 MARCOS HENRIQUE SILVA ARAGÃO - DISSERTAÇÃO (PPGECA) 2017.pdf: 4540927 bytes, checksum: d25472de237d384e285a54b061998c1e (MD5) Previous issue date: 2017-08 / Capes / No processo de produção do álcool são gerados até 15 litros de vinhoto por litro de álcool produzido. O vinhoto é um rejeito caracterizado por apresentar elevadas concentrações de material orgânico e nutrientes. Essas características fazem do vinhoto um substrato adequado para a produção de proteína unicelular (PUC) em sistemas de lodos ativados mediante a produção de excessiva de biomassa bacteriana, que é constituída por cerca de 60 -70 % de proteínas, podendo ser empregada como ração animal. O lodo do tipo PAO, produzido em sistemas EBPR, é caracterizado por ter uma taxa de decaimento bacteriano notadamente inferior em comparação aos microrganismos ordinários de lodos ativados. Isso representa uma vantagem para a produção de PUC, tanto pela manutenção de uma quantidade maior de biomassa, quanto pelos menores requisitos de oxigênio para a oxidação das células mortas. O desenvolvimento desses microrganismos no lodo exige uma mudança operacional no regime de aeração, introduzindo uma fase anaeróbia em cada batelada, na qual é realizada a alimentação com o substrato. Este trabalho teve como objetivos: avaliar e comparar a produção máxima de PUC a partir do vinhoto em sistemas de lodos ativados em função da idade de lodo tanto para os organismos heterotróficos ordinários (OHO) como para organismos acumuladores de polifosfato (PAO), determinar a eficiência de remoção de material orgânico e de nutrientes, a determinação das composições dos lodos e a determinação das principais constantes cinéticas e estequiométricas de cada um deles. Foi aplicada a respirometria como ferramenta para a caracterização cinética do consumo do material orgânico e adotada a teoria de lodos ativados proposta por van Haandel e Marais (1999). Foram operados e monitorados dois reatores em bateladas sequenciais com idade de lodo de 8 dias, sendo um deles submetido a uma interrupção no regime de aeração, introduzindo uma fase anaeróbia no início de cada batelada visando ao desenvolvimento de um lodo do tipo PAO. Os resultados obtidos demonstraram que houve uma produção satisfatória de lodo em ambos os reatores, bem como uma elevada remoção de material orgânico no efluente. Não se observou uma remoção em excesso de fósforo ou uma elevada fração de fósforo incorporada ao lodo produzido no sistema submetido à modificação no regime de aeração, além disso não foi observada uma produção mais elevada de lodo nesse sistema, indicando que o lodo PAO não foi produzido. A determinação do comportamento cinético dos lodos demonstrou que existe um elevado potencial de produção de PUC em sistemas lodos ativados tratando vinhoto, embora essa produção seja limitada pela capacidade de oxigenação dos aeradores e esteja condicionada ao avanço desses equipamentos. / In the process of producing the alcohol, up to 15 liters of vinasse per liter of alcohol produced are generated. The vinasse is a reject characterized by high concentrations of organic material and nutrients. These characteristics make vinasse a suitable substrate for the production of single cell protein (SCP) in activated sludge systems through the production of excessive bacterial biomass, which consists of about 60-70% of proteins, can be used as animal feed. The PAO-type sludge, produced in EBPR systems, is characterized by having a markedly lower bacterial decay rate compared to ordinary activated sludge microorganisms. This represents an advantage for the production of PUC, both by maintaining a higher amount of biomass and by lower oxygen requirements for the oxidation of dead cells. The development of these microorganisms in the sludge requires an operational change in the aeration regime, introducing an anaerobic phase in each batch, in which the feeding with the substrate is carried out. The objective of this work was to evaluate and compare the maximum production of SCP from vinasse as a function of sludge age in activated sludge systems for both ordinary heterotrophic organisms (OHO) and polyphosphate accumulating organisms (PAO). Efficiency of removal of organic matter and nutrients, determination of the sludge compositions and determination of the main kinetic and stoichiometric constants of each of them. It was applied the respirometry as a tool for the kinetic characterization of organic material consumption and adopted the theory of activated sludge proposed by van Haandel and Marais (1999). Two reactors were operated and monitored in sequential batches with a sludge age of 8 days, one of them being submitted to an interruption in the aeration regime, introducing an anaerobic phase at the beginning of each batch, aiming at the development of a PAO type sludge. The results showed that there was a satisfactory production of sludge in both reactors, as well as a high removal of organic material in the effluent. There was no excess removal of phosphorus or a high fraction of phosphorus incorporated into the sludge produced in the system submitted to the modification in the aeration regime, in addition no higher sludge production was observed in this system, indicating that the PAO sludge was not produced. The determination of the kinetic behavior of the sludge showed that there is a high potential in the PUC production in activated sludge treatment systems, although this production is limited by the oxygenation capacity of the aerators and is conditioned to the advance of these equipment.

Fish has for a long time been a very important source of protein for human kind and with the world population at an all-time high, 7.5 billion and rapidly growing, the demand for fish as a food source is also at an all-time high and rapidly increasing. This has in turn led to overexploitation of many of the fish stocks of the world ocean’s and in many cases to depletion of fish stocks. The demand for sustainable food sources and sustainable usage of the world ocean’s fish stocks is therefore a subject with great deal of interest today. Much of the fish caught today are used for production of fish meal for usage as fish food at fish farms, which also increases the depletion of fish stocks around the globe. One way of dealing with this problem is to replace the fish meal as protein source in fish feed with protein from agricultural crops which in many cases are done today by usage of soy bean protein. This however poses another problem as the agricultural crops take up vast amount of land, in many cases obtained by diminishing the rainforests in the area. Another usage for the soybean would be as a direct human food source. Agricultural products are also dependent on environmental conditions to ensure reasonable production. The problems related to production of fish meal and soy has sparked the idea of using microorganisms for production of Single cell protein for usage as protein source in fish feed. Single cell protein can be produced in closed fermentation vessels and can be produced at a controlled rate and under controlled manners, while taking up negligible land space. During this thesis, the production of single cell protein from spent sulfite liquor using the filamentous fungi Paecilomyces variotii was examined. The aim of the project was to examine the effect of cultivation parameters (i.e., pH, temperature and nutrients) on the production of biomass as well as the protein content of the biomass. The correlation of the biomass growth and protein content have also been examined. The project was carried out by performing several experiment cultivations using spent sulfite liquor provided by Domsjö Fabriker in Örnsköldsvik. This process enables the utilization of a residual stream from the pulp industry which gives this process a huge environmental upside compared to similar processes as for example the commercial production of Quorn (a Single cell based food product) which utilizes pure glucose. The results showed that the protein content will steadily decrease as the biomass production increases hence it is desirable to keep the cultivation time at a minimum while maximizing biomass production during that time frame. It also points towards that the highest protein content is present in the young cell mass. The key conclusion from this thesis is however that it is possible to lower the pH of the cultivation from pH 6 down to pH 4.5 while still maintaining the biomass production and increasing the protein content. The highest obtained protein content was 62.7% at pH 4.5. The high protein content might be due to a slightly longer lag phase in the beginning of the cultivation which yields a higher number of younger cells in the final broth thus increasing protein content. Running the process at a lower pH is a huge advantage for industrial implementation as this on large scale means significant lower amounts of chemicals needed for pH adjusting of the spent sulfite liquor which renders the process much more economical. This is because pH adjustment today is one of the most costly process steps in the production of bioethanol from spent sulfite liquor.