Orientador: Marcela Aparecida Guerreiro Machado / Resumo: A microalga Chlorella possui ácidos graxos poli-insaturados, vitaminas e alto conteúdo proteico. Desse modo, estudos recentes têm explorado o uso de microalgas para obtenção de lipídios, principalmente os de maior valor comercial como o ácido ɣ-linolênico. Estre trabalho objetivou propor um método de cultivo para a microalga Chlorella vulgaris e extração do óleo. Os experimentos foram realizados em duplicata de acordo com o arranjo ortogonal de Taguchi, a partir da qual foram organizadas duas etapas: na primeira foram viabilizados cultivos nos quais os fatores de interesse pudessem ser avaliados em dois níveis de operação: Aeração (1,67 e 3,33L/min), NO3 (0,25 e 0,50 g/L), PO4 -3 (0,35 e 0,65 g/L), NaHCO3 (0,25 e 0,50 g/L), intensidade de luz (0,85 e 14,50 K luz) e fotoperíodo (12 e 24 h). Em tal etapa correu também a colheita, floculação, secagem e moagem da biomassa. Na segunda etapa foram realizadas as extrações lipídicas de acordo com e uso de solventes como o clorofórmio e metanol, utilizou-se também o banho de ultrassom para as extrações dos teores lipídicos. Obteve melhor resultado a concentração de biomassa algal, a qual atingiu a concentração de nitrato (0,50 g/L), concentração de bicarbonato de sódio (0,50 g/L), fosfato (0,65 g/L) e fotoperíodo de 12 em 12hs. O melhor resultado para a extração lipídica ocorreu conforme a metodologia de Zorn (2017), com aproximadamente 2,5% mais eficiente e obteve o seguinte resultado para os fatores, no nível alto são: aeração (3,33... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The micro-algae Chlorella has polyunsaturated fatty acids, vitamins and high protein content. Thus, recent studies have explored the use of microalgae to obtain lipids, especially those with higher commercial value such as β-linolenic acid. The objective of this work was to propose a method of cultivation for the microalga Chlorella vulgaris and extraction of the oil. The experiments were carried out in duplicate according to Taguchi's orthogonal arrangement, from which two stages were organized: in the first, cultures were feasible in which the factors of interest could be evaluated at two levels of operation: aeration (1.67 e 3.33L/min), PO4 -3 (0.35 and 0.65 g / L), NaHCO 3 (0.25 and 0.50 g / L), NO 3 (0.25 and 0.50 g / L) , light intensity (0.85 and 14.50 K light) and photoperiod (12 and 24 h). In this stage also the harvest, flocculation, drying and milling of the biomass took place. In the second stage, the lipid extractions were performed according to the use of solvents such as chloroform and methanol, and the ultrasonic bath was used to extract the lipid contents. The best results were the algal biomass concentration, which reached the concentration of nitrate (0.50 g / L), sodium bicarbonate (0.50 g / L), phosphate (0.65 g / L) and photoperiod every 12 hours The best result for the lipid extraction occurred according to the Zorn (2017) methodology, with approximately 2.5% more efficient and obtained the following result for the factors, at the high level are: aerati... (Complete abstract click electronic access below) / Mestre

With greenhouse gas emissions (of which CO2 is the major component) being a major environmental concern, mitigation of those emissions is becoming increasingly imperative. The ability to use a fast growing, photosynthetic organism like microalgae that can survive primarily on nutrients such as sunlight and air (with increased CO2 levels) makes it a desirable agent for CO2 sequestration. The primary goal of this project is the engineering of the sequestration of CO2 using the cultivation of the microalgae species <i>Chlorella vulgaris</i>. Secondary goals of the project are the exploration and development of valuable by-products of the cultivation and the determination of whether utilizing microalgae to capture CO2 could be integrated economically into an industrial facility.<p> The batch growth kinetics of the photosynthetic algal species <i>C. vulgaris</i> were investigated using a well-mixed stirred bioreactor. The growth rate was found to increase as the dissolved CO2 increased to 150 mg/L (10% CO2 by volume in the gas), but fell dramatically at higher concentrations. Increasing the radiant flux also increased growth rate. With a radiant flux of 32.3 mW falling directly on the 500 mL culture media, the growth rate reached up to 3.6 mg of cells/L-h. Both pH variation (5.5 - 7.0) and mass transfer rate of CO2 (KLa between 6 h-1 and 17 h-1) had little effect on growth rate.<p> The operation of continuously stirred tank bioreactors (CSTBs) at minimum cost is a major concern for operators. In this work, a CSTB design strategy is presented where impeller stirring speed and aeration rate are optimized to meet the oxygen demand of growing cells, simultaneously minimizing the capital and operating cost. The effect of microbial species, ions in the culture medium, impeller style, as well as changing CSTB size and biomass input density on the optimum operating conditions, is examined. A study of the effects of various parameters on the CSTB design is shown.<p> Using the kinetic data collected in the batch growth study, a novel external loop airlift photobioreactor (ELAPB) was designed and tested. A model was developed for <i>C. vulgaris</i> growth in the ELAPB that incorporated growth behaviour, light attenuation, mass transfer, and fluid dynamics. The model predicts biomass accumulation, light penetration, and transient CO2 concentrations, and compares predictions to experimental data for radiant fluxes of 0.075 1.15 W/m2 and 0 20% CO2 enrichment of feed air, with a 10% average error. The effect of radiant flux and CO2 concentration is presented with discussion of radial and vertical profiles along the column. For a fed-batch culture at a biomass density of 170 mg/L, the penetration of the radiant flux was found to decrease by 50% within the first 1 cm, and 75% at 2 cm. Theoretical optimum growth conditions are determined to be 0.30 W/m2 and 6% CO2 enrichment of inlet feed air.<p> The algal culture was observed to be a workable electron acceptor in a cathodic half cell. A net potential difference of 70 mV was achieved between the growing <i>C. vulgaris</i> culture acting as a cathode and a 0.02 M potassium ferrocyanide anodic half cell. Surge current and power levels of 1.0 µA/mg of cell dry weight and 2.7 mW/m2 of cathode surface area were measured between these two half cells. The recently developed photosynthetic cathode was also coupled to a fermentative anode to produce a completely microbial fuel cell. Loading effects and the effect of changing culture conditions on fuel cell operation are reported. The maximum power output measured was 0.95 mW/ m2 at 90 V and 5000 ohms. A significant increase in this output is achieved with the addition of supplemental glucose to the anodic half cell and the enrichment of the feed air bubbled into the cathodic half cell with 10% CO2.<p> Two economic feasibility studies were performed on the integration of ELAPBs into an industrial facility. These integration studies operated the ELAPBs continuously as biocathodes in coupled microbial fuel cells (MFCs) that capture CO2 from an existing 130 million L/yr bioethanol plant, while generating electrical power and yielding oil for biodiesel to provide operational revenue to offset costs. The anodes for the coupled MFCs are the existing yeast batch fermentors, and the CO2 to be sequestered comes from the existing bioethanol production. Two different design schemes were evaluated, in both cases the maximum profit was achieved with the maximum number of tall columns operated in parallel. The first design evaluated a batch bioethanol facility with off-site oil processing, and the economic feasibility is demonstrated by the positive Net Present Worth achieved over the 20 year life of the plant, at a 10% rate of return on investment. The second design, for a continuous bioethanol operation, processes both oil and algae biomass on-site, but the economics of this second process are only positive (Internal Rate of Return 9.93%.) if the government provides financial assistance in the form of generous carbon credits (a speculative $100 per tonne of CO2 not yet attained) and a 25% capital equipment grant.

With greenhouse gas emissions (of which CO2 is the major component) being a major environmental concern, mitigation of those emissions is becoming increasingly imperative. The ability to use a fast growing, photosynthetic organism like microalgae that can survive primarily on nutrients such as sunlight and air (with increased CO2 levels) makes it a desirable agent for CO2 sequestration. The primary goal of this project is the engineering of the sequestration of CO2 using the cultivation of the microalgae species <i>Chlorella vulgaris</i>. Secondary goals of the project are the exploration and development of valuable by-products of the cultivation and the determination of whether utilizing microalgae to capture CO2 could be integrated economically into an industrial facility.<p> The batch growth kinetics of the photosynthetic algal species <i>C. vulgaris</i> were investigated using a well-mixed stirred bioreactor. The growth rate was found to increase as the dissolved CO2 increased to 150 mg/L (10% CO2 by volume in the gas), but fell dramatically at higher concentrations. Increasing the radiant flux also increased growth rate. With a radiant flux of 32.3 mW falling directly on the 500 mL culture media, the growth rate reached up to 3.6 mg of cells/L-h. Both pH variation (5.5 - 7.0) and mass transfer rate of CO2 (KLa between 6 h-1 and 17 h-1) had little effect on growth rate.<p> The operation of continuously stirred tank bioreactors (CSTBs) at minimum cost is a major concern for operators. In this work, a CSTB design strategy is presented where impeller stirring speed and aeration rate are optimized to meet the oxygen demand of growing cells, simultaneously minimizing the capital and operating cost. The effect of microbial species, ions in the culture medium, impeller style, as well as changing CSTB size and biomass input density on the optimum operating conditions, is examined. A study of the effects of various parameters on the CSTB design is shown.<p> Using the kinetic data collected in the batch growth study, a novel external loop airlift photobioreactor (ELAPB) was designed and tested. A model was developed for <i>C. vulgaris</i> growth in the ELAPB that incorporated growth behaviour, light attenuation, mass transfer, and fluid dynamics. The model predicts biomass accumulation, light penetration, and transient CO2 concentrations, and compares predictions to experimental data for radiant fluxes of 0.075 1.15 W/m2 and 0 20% CO2 enrichment of feed air, with a 10% average error. The effect of radiant flux and CO2 concentration is presented with discussion of radial and vertical profiles along the column. For a fed-batch culture at a biomass density of 170 mg/L, the penetration of the radiant flux was found to decrease by 50% within the first 1 cm, and 75% at 2 cm. Theoretical optimum growth conditions are determined to be 0.30 W/m2 and 6% CO2 enrichment of inlet feed air.<p> The algal culture was observed to be a workable electron acceptor in a cathodic half cell. A net potential difference of 70 mV was achieved between the growing <i>C. vulgaris</i> culture acting as a cathode and a 0.02 M potassium ferrocyanide anodic half cell. Surge current and power levels of 1.0 µA/mg of cell dry weight and 2.7 mW/m2 of cathode surface area were measured between these two half cells. The recently developed photosynthetic cathode was also coupled to a fermentative anode to produce a completely microbial fuel cell. Loading effects and the effect of changing culture conditions on fuel cell operation are reported. The maximum power output measured was 0.95 mW/ m2 at 90 V and 5000 ohms. A significant increase in this output is achieved with the addition of supplemental glucose to the anodic half cell and the enrichment of the feed air bubbled into the cathodic half cell with 10% CO2.<p> Two economic feasibility studies were performed on the integration of ELAPBs into an industrial facility. These integration studies operated the ELAPBs continuously as biocathodes in coupled microbial fuel cells (MFCs) that capture CO2 from an existing 130 million L/yr bioethanol plant, while generating electrical power and yielding oil for biodiesel to provide operational revenue to offset costs. The anodes for the coupled MFCs are the existing yeast batch fermentors, and the CO2 to be sequestered comes from the existing bioethanol production. Two different design schemes were evaluated, in both cases the maximum profit was achieved with the maximum number of tall columns operated in parallel. The first design evaluated a batch bioethanol facility with off-site oil processing, and the economic feasibility is demonstrated by the positive Net Present Worth achieved over the 20 year life of the plant, at a 10% rate of return on investment. The second design, for a continuous bioethanol operation, processes both oil and algae biomass on-site, but the economics of this second process are only positive (Internal Rate of Return 9.93%.) if the government provides financial assistance in the form of generous carbon credits (a speculative $100 per tonne of CO2 not yet attained) and a 25% capital equipment grant.

The use of microalgae culture to convert CO2 from power plant flue gases into biomass that are readily converted into biofuels offers a new frame of opportunities to enhance, compliment or replace fossil-fuel-use. Apart from being renewable, microalgae also have the capacity to utilise materials from a variety of wastewater and the ability to yield both liquid and gaseous biofuels. However, the processes of cultivation, incorporation of a production system for power plant waste flue gas use, algae harvesting, and oil extraction from the biomass have many challenges. Using SimaPro software, Life cycle Assessment (LCA) of the challenges limiting the microalgae (Chlorella vulgaris) biofuel production process was performed to study algae-based pathway for producing biofuels. Attention was paid to material use, energy consumed and the environmental burdens associated with the production processes. The goal was to determine the weak spots within the production system and identify changes in particular data-set that can lead to and lower material use, energy consumption and lower environmental impacts than the baseline microalgae biofuel production system. The analysis considered a hypothetical transesterification and Anaerobic Digestion (AD) transformation of algae-to- biofuel process. Life cycle Inventory (LCI) characterisation results of the baseline biodiesel (BD) transesterification scenario indicates that heating to get the biomass to 90% DWB accounts for 64% of the total input energy, while electrical energy and fertilizer obligations represents 19% and 16% respectively. Also, Life Cycle Impact Assessment (LCIA) results of the baseline BD production scenario show high proportional contribution of electricity and heat energy obligations for most impact categories considered relative to other resources. This is attributed to the concentration/drying requirement of algae biomass in order to ease downstream processes of lipid extraction and subsequent transesterification of extracted lipids into BD. Thus, four prospective alternative production scenarios were successfully characterised to evaluate the extent of their impact scenarios on the production system with regards to lowering material use, lower energy consumption and lower environmental burdens than the standard algae biofuel production system. A 55.3% reduction in mineral use obligation was evaluated as the most significant impact reduction due to the integration of 100% recycling of production harvest water for the AD production system. Recycling also saw water demand reduced from 3726 kg (freshwater).kgBD- 1 to 591kg (freshwater).kgBD- 1 after accounting for evaporative losses/biomass drying for the BD transesterification production process. Also, the use of wastewater/sea water as alternative growth media for the BD production system, indicated potential savings of: 4.2 MJ (11.8%) in electricity/heat obligation, 10.7% reductions for climate change impact, and 87% offset in mineral use requirement relative to the baseline production system. Likewise, LCIA characterisation comparison results comparing the baseline production scenarios with that of a set-up with co-product economic allocation consideration show very interesting outcomes. Indicating -12 MJ surplus (-33%) reductions for fossil fuels resource use impact category, 52.7% impact reductions for mineral use impact and 56.6% reductions for land use impact categories relative to the baseline BD production process model. These results show the importance of allocation consideration to LCA as a decision support tool. Overall, process improvements that are needed to optimise economic viability also improve the life cycle environmental impacts or sustainability of the production systems. Results obtained have been observed to agree reasonably with Monte Carlo sensitivity analysis, with the production scenario proposing the exploitation of wastewater/sea water to culture algae biomass offering the best result outcome. This study may have implications for additional resources such as production facility and its construction process, feedstock processing logistics and transport infrastructure which are excluded. Future LCA study will require extensive consideration of these additional resources such as: facility size and its construction, better engineering data for water transfer, combined heat and power plant efficiency estimates and the fate of long-term emissions such as organic nitrogen in the AD digestate. Conclusions were drawn and suggestions proffered for further study.

Uran kann sowohl durch geogene als auch anthropogene Vorgänge in die Umwelt gelangen. Dazu zählen natürliche Uranerzvorkommen und deren Leaching sowie die Auswaschung von Uran aus den Hinterlassenschaften des ehemaligen Uranerzbergbaus. Die Aufklärung des Verhaltens von Uran in der Geo- und Biosphäre ist für eine Risikoabschätzung des Migrationsverhaltens von Radionukliden in der Umwelt notwendig. Algen sind in der Natur weit verbreitet und die wichtigste Organismengruppe in den aquatischen Lebensräumen. Durch ihre ubiquitäre Verbreitung in der Natur ist ihr Einfluss auf das Migrationsverhalten von Uran in der Umwelt von grundlegendem Interesse z.B. um effektive und wirtschaftliche Remediationsstrategien für Wässer zu entwickeln. Außerdem stehen Algen am Beginn der Nahrungskette und spielen eine wirtschaftlich relevante Rolle als Nahrung beziehungsweise Nahrungsergänzungsmittel. Die Möglichkeit des Transfers von algengebundenem Uran entlang der Nahrungskette könnte eine ernsthafte Gesundheitsgefahr für den Menschen darstellen. Das Ziel dieser Arbeit war die quantitative und strukturelle Charakterisierung der Wechselwirkung zwischen Uran(VI) und der Grünalge Chlorella vulgaris im umweltrelevanten Konzentrations- und pH-Wertbereich unter besonderer Berücksichtigung der Stoffwechselaktivität. Die in dieser Arbeit erzielten Ergebnisse der Sorptionsexperimente zeigen deutlich den maßgeblichen Einfluss des Stoffwechselstatus von Chlorella auf die Wechselwirkung mit Uran. So kann in Gegenwart von umweltrelevanten Urankonzentrationen eine Remobilisierung von zuvor passiv gebundenem Uran durch die stoffwechselaktiven Algen erfolgen. Die in Abhängigkeit von der Stoffwechselaktivität, der Urankonzentration und dem pH-Wert mit den Algenzellen gebildeten Uran(VI)-Komplexe wurden strukturell mit Hilfe der spektroskopischen Methoden TRLF-, EXAFS- und ATR-FTIR-Spektroskopie charakterisiert. Mittels TEM konnte Uran in Form von 30-70 nm großen nadelförmigen Ablagerungen in der Zellwand der lebende Algenzellen nachgewiesen werden. Die in dieser Arbeit erhaltenen Ergebnisse leisten einen wichtigen Beitrag zur Vorhersage des Migrationsverhaltens von Uran unter umweltrelevanten Bedingungen und der radiologischen Risikobewertung von geogen und anthropogen auftretendem Uran.

Neste trabalho é proposto o desenvolvimento de um processo para a biofixação do dióxido de carbono através do uso de microalgas. Para o cultivo desses microrganismos foram utilizados fotobiorreatores do tipo airlift. Nos cultivos foram avaliadas espécies de microalgas (Chlorella sp. e Chlorella minutissima), influência da intensidade luminosa (2.200 a 24.500 lx), concentração salina (28 a 40 g.L-1) e temperatura (25 °C a 35 °C) sobre a concentração de biomassa, velocidade específica de crescimento, produtividade de biomassa, biofixação de CO2, conteúdo lipídico e carotenoides totais. Também foi realizada a identificação dos carotenoides. A microalga escolhida para os testes em fotobiorreatores foi a C. minutissima. A intensidade luminosa que apresentou os melhores resultados foi a de 17.000 lx. A temperatura mostrou possuir influência significativa na concentração de biomassa, na velocidade específica de crescimento e na biofixação de carbono, mas a concentração salina influenciou apenas a velocidade específica de crescimento. A temperatura de 25 °C apresentou as maiores produtividade de biomassa (0,094 g.L-1.d-1) e concentração de biomassa (0,43 g.L-1), independente da concentração salina, e as maiores velocidade específica de crescimento (0,81 d-1) e biofixação de CO2 (12 gCO2.m-3.h-1), na concentração salina de 37 g.L-1. O conteúdo lipídico médio das microalgas foi de 13,2 % e os carotenoides totais apresentaram teor de 0,25 % do peso seco das microalgas em todas as condições de concentração salina e temperatura testadas. Foi possível identificar a produção dos carotenoides luteína, zeaxantina e β-caroteno. / The present study proposes the development of a process for carbon dioxide biofixation through the use of microalgae. Flat-plate airlift photobioreactors were used. Microalgaes species (Chlorella sp. and Chlorella minutissima) and the influence of light intensity (2,200 to 24,000 lx), salt concentration (28 to 40 g.L-1) and temperature (25 to 35 °C) over biomass concentration, specific growth rate, biomass productivity, CO2 biofixation rate, lipid content and total carotenoids content were evaluated. The identification of the carotenoids was performed. C. minutissima showed the best performance in shaker and was chosen for the tests in photobioreactor. The light intensity of 17,000 lx presented the best results. The temperature showed to have significant influence over biomass concentration, specific growth rate and CO2 biofixation rate, but the salt concentration only affected the specific growth rate. The temperature of 25 °C allowed the highest biomass productivity (0.094 g.L-1.d-1) and biomass concentration (0.43 g.L-1), independent of salt concentration, and the highest specific growth rate (0.81 d-1) and CO2 biofixation rate (12 gCO2.m-3.h-1) at the salt concentration of 37 g.L-1. The average lipid content of the microalgae was 13.2 % and the total carotenoids content were about 0.25 % of the cell dry weight at all temperatures and salt concentrations tested. It was possible to identify the production of the carotenoids lutein, zeaxanthin and β-carotene.

Submitted by Juliana Laiate (jlaiatetana@gmail.com) on 2018-10-04T19:13:17Z No. of bitstreams: 1 Juliana Laiate_Tese Doutorado_Correção Biblioteca_Versão Final.pdf: 5872852 bytes, checksum: c627770397e856b9d9aba7d0f399f835 (MD5) / Approved for entry into archive by Pamella Benevides Gonçalves null (pamella@feg.unesp.br) on 2018-10-05T13:30:56Z (GMT) No. of bitstreams: 1 laiate_j_dr_guara.pdf: 5872852 bytes, checksum: c627770397e856b9d9aba7d0f399f835 (MD5) / Made available in DSpace on 2018-10-05T13:30:56Z (GMT). No. of bitstreams: 1 laiate_j_dr_guara.pdf: 5872852 bytes, checksum: c627770397e856b9d9aba7d0f399f835 (MD5) Previous issue date: 2018-08-31 / Outra / O interesse na diversificação da matriz energética por meio de fontes de energias renováveis tem promovido mudanças no âmbito nacional e internacional, em setores acadêmicos, industriais, sociais e governamentais com foco no desenvolvimento de processos biotecnológicos baseados nos três pilares da sustentabilidade: ambiental, social e econômico. Frente a esse contexto, as microalgas são consideradas uma fonte promissora alternativa para a produção de biocombustíveis por apresentarem alta taxa de crescimento e de concentração de lipídeos. O presente trabalho realizou um estudo do processo de cultivo da microalga marinha Chlorella minutíssima por meio da ferramenta DOE – Design of Experiments para maximização da sua concentração microalgal e do teor lipídico simultaneamente, além de sua caracterização termoquímica para aplicação no processo de gaseificação. O cultivo ocorreu em fotobiorreator tubular descontínuo do tipo coluna de bolhas seguindo o arranjo ortogonal L8 de Taguchi, e as variáveis de processo foram fluxo de CO2 na alimentação, concentração de nitrato, concentração de fosfato, suplementação, temperatura e salinidade. A investigação demonstrou que a concentração de nitrato e fosfato, o fluxo de gás carbônico, a suplementação do meio e a temperatura são fatores influentes na concentração microalgal e no teor lipídico, sendo a melhor configuração para maximização conjunta, utilizando-se a função desirability, com o nitrato operando em nível alto e os demais fatores em nível baixo. A análise elementar forneceu teores condizentes com alguns relatos da literatura, a termogravimetria auxiliou na identificação das faixas de temperatura com maior taxa de decomposição (300 °C a 400 °C), e o poder calorífico superior da biomassa encontrado se demonstrou atrativo para a produção de syngas quando comparado ao de materiais como casca de arroz e lascas de eucalipto já empregados na gaseificação em maior escala. Dessa forma, a metodologia empregada e os resultados apresentados neste trabalho podem auxiliar na busca da viabilização econômica do cultivo, em grande escala, da microalga marinha Chlorella minutíssima para produção de biocombustível pelo processo de gaseificação. / The interest in diversifying the energy matrix through renewable energy sources has promoted changes at the national and international levels in academic, industrial, social and governmental sectors, focused on the development of biotechnological processes based on the three pillars of sustainability: environmental, social and economic. In this context, microalgae are considered a strong promising alternative source for the production of biofuels due to their high rate of growth and lipid concentration. The present work carried out a study of the cultivation process of the marine microalgae Chlorella minutíssima using DOE – Design of Experiments to maximize the cell growth and the lipid content simultaneously, besides its thermochemical characterization for application in the gasification process. Cultivation occurred in a discontinuous-tubular photobioreactor, bubble-column type, following Taguchi L8 Orthogonal Array design, and the process variables were: CO2 feed flow rate, nitrate concentration, phosphate concentration, supplementation (metals and vitamins), temperature and salinity. Results showed that the nitrate and phosphate concentration, CO2 feed flow, supplements and temperature are influential factors in the cell growth and lipid content, and the best configuration presenting good results for simultaneous maximization, using desirability, was nitrate operating at high level and the other factors at low level. Ultimate analysis provided levels consistent with some reports in the literature. Thermogravimetric analysis identified the temperature ranges with the highest decomposition rate (300 °C to 400 °C). The higher heating value of the biomass found was attractive for the production of syngas when compared to raw materials such as rice husk and eucalyptus chips already used in the larger scale gasification. In face of, the methodology employed and the results presented in this work can be useful in seeking economic feasibility of the large-scale cultivation of Chlorella minutíssima for the production of biofuels by the gasification process. / PRH48 - ANP

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência dos Alimentos. / Made available in DSpace on 2012-10-26T12:33:48Z (GMT). No. of bitstreams: 1 303103.pdf: 2097012 bytes, checksum: 723a0292e37771449437e286b09bf76c (MD5) / O objetivo deste trabalho foi avaliar as propriedades físicoquímicas da biomassa de Chlorella vulgaris em um meio à base de concentrado de dessalinização proveniente do processo de dessalinização de águas subterrâneas na comunidade de Uruçu, no município de São João do Cariri # Paraíba. O concentrado de dessalinização possui alta concentração de sais minerais (cloretos, carbonatos e bicarbonatos) que se reincorporado ao solo podem causar o fenômeno de eritrofização, erosão e salinização do solo. Por isso, diversas pesquisas têm sido realizadas de modo a encontrar soluções de reciclagem deste concentrado. Entre estas pesquisas, o cultivo de microalgas para a produção de biomassa com finalidade biotecnológica. O cultivo autotrófico de Chlorella vulgaris foi inicialmente mantido em Bold Basal Medium (BBM). Para fazer uso do concentrado de dessalinização, realizou-se a caracterização físico-química, determinada segundo metodologia descrita no Standard Methods for the Examination of Water and Wastewater - APHA (2005). Com a aplicação da Metodologia de Superfície de Resposta foi possível avaliar o efeito de três nutrientes: NaNO3, KH2PO4 e solução micronutriente nas concentrações estabelecidas pelo planejamento experimental. O meio de cultura proposto para maximizar a produção de biomassa de Chlorella vulgaris consistiu de 0,09 g L-1 de NaNO3, 0,014 g L-1 de KH2PO4 e 0,20 mL L-1 de solução micronutriente. Otimizado, o novo meio de cultura serviu de base para realizar um cultivo contínuo em fotobiorreator, com o objetivo de avaliar os teores de proteína e lipídeos totais que resultaram em 43,0 % e 9,5 %, respectivamente. A espécie Chlorella vulgaris apresentou predominância em três ácidos graxos: ácido linolênico (C18:3n3c); ácido palmítico (C16:0) e ácido linoléico (C18:2n6c), cujas proporções foram de 20,9 %; 20,5 %; 18,8 %, respectivamente. Estes ácidos graxos têm entre 14 e 18 carbonos na cadeia, sendo predominantemente poli-insaturados (39,7 %), saturados (25,7 %) e monoinsaturados (14,0 %). Adiante, com o novo meio de cultura realizou-se um cultivo sucessivo em biorreatores cônicos, a fim de avaliar a capacidade de reuso da solução contendo concentrado de dessalinização. Quatro foram os ciclos de cultivo. Para cada ciclo de cultivo, os valores de biomassa resultaram em 1° ciclo, 1,55 g L-1; 2° ciclo, 0,96 g L-1; 3° ciclo, 0,62 g L-1 e 4° ciclo em 0,42 g L-1. Neste trabalho foi observado que a biomassa de Chlorella vulgaris é fonte de ácidos graxos essenciais -3 e -6. Por fim, obteve-se um meio de cultivo alternativo e de baixo custo para o crescimento de Chlorella vulgaris / The aim of this study was to optimize a medium culture for the microalgae Chlorella vulgaris using the concentrate from desalination process of groundwater by reverse osmosis from a water desalination plant in the community of Uruçu, in São João do Cariri # Paraíba/Brazil. The desalination concentrate contains high mineral concentrations such as chlorides, carbonates and bicarbonates, which may increase the risk of soil salinization in the case of being reincorporated in the soil. Therefore, many studies have been performed in order to find solutions for recycling this waste. Autotrophic cultivation of Chlorella vulgaris was initially kept in Bold Basal Medium (BBM). In order to use the desalination concentrate, the physical-chemical parameters were determined according to the Standard Methods for the Examination of Water and Wastewater # APHA (2005). The Response Surface Methodology was used in order to evaluate the effect of three nutrients: NaNO3, KH2PO4 and micronutrient solution as determined by the experimental design. The culture medium proposed for maximizing the biomass production of Chlorella vulgaris consists of 0.09 g L-1 of NaNO3; 0.014 g L-1 of KH2PO4 and 0.20 mL L-1 of micronutrient solution. Once optimized, the new culture medium was used as the basis for performing a continuous cultivation in photobioreactors, in which the total protein and lipids resulted in 43 % and 9.5 %, respectively. The Chlorella vulgaris species produced, predominantly, three fatty acids: linolenic acid (C18:3n3c); palmitic acid (C16:0) and linolenic acid (C18:2n6c), whose contents were 20.9 %; 20.5 %; 18.8 %, respectively. These fatty acids present 14 to 18 carbons in the carbon chain, being mainly polyunsaturated (39.7 %), saturated (25.7 %) and monounsaturated (14.0 %). With the optimized culture medium, then, a continuous cultivation in bioreactors was carried out. After each cultivation period, the biomass was collected and the culture medium was used in the further cultivation cycles in order to evaluate the ability of reusing the solution containing the desalination concentrate. Four cultivation cycles were performed. The values of biomass resulted in 1° cycle, 1.55 g L-1; 2° cycle, 0.96 g L-1; 3° cycle, 0.62 g L-1 and 4° cycle, 0.42 g L-1. In this work, it was observed that the biomass of Chlorella vulgaris is a potential source of essential fatty acids -3 e -6. Finally, it was possible to obtain an alternative and low cost culture medium for the growth of Chlorella vulgaris

Neste trabalho é proposto o desenvolvimento de um processo para a biofixação do dióxido de carbono através do uso de microalgas. Para o cultivo desses microrganismos foram utilizados fotobiorreatores do tipo airlift. Nos cultivos foram avaliadas espécies de microalgas (Chlorella sp. e Chlorella minutissima), influência da intensidade luminosa (2.200 a 24.500 lx), concentração salina (28 a 40 g.L-1) e temperatura (25 °C a 35 °C) sobre a concentração de biomassa, velocidade específica de crescimento, produtividade de biomassa, biofixação de CO2, conteúdo lipídico e carotenoides totais. Também foi realizada a identificação dos carotenoides. A microalga escolhida para os testes em fotobiorreatores foi a C. minutissima. A intensidade luminosa que apresentou os melhores resultados foi a de 17.000 lx. A temperatura mostrou possuir influência significativa na concentração de biomassa, na velocidade específica de crescimento e na biofixação de carbono, mas a concentração salina influenciou apenas a velocidade específica de crescimento. A temperatura de 25 °C apresentou as maiores produtividade de biomassa (0,094 g.L-1.d-1) e concentração de biomassa (0,43 g.L-1), independente da concentração salina, e as maiores velocidade específica de crescimento (0,81 d-1) e biofixação de CO2 (12 gCO2.m-3.h-1), na concentração salina de 37 g.L-1. O conteúdo lipídico médio das microalgas foi de 13,2 % e os carotenoides totais apresentaram teor de 0,25 % do peso seco das microalgas em todas as condições de concentração salina e temperatura testadas. Foi possível identificar a produção dos carotenoides luteína, zeaxantina e β-caroteno. / The present study proposes the development of a process for carbon dioxide biofixation through the use of microalgae. Flat-plate airlift photobioreactors were used. Microalgaes species (Chlorella sp. and Chlorella minutissima) and the influence of light intensity (2,200 to 24,000 lx), salt concentration (28 to 40 g.L-1) and temperature (25 to 35 °C) over biomass concentration, specific growth rate, biomass productivity, CO2 biofixation rate, lipid content and total carotenoids content were evaluated. The identification of the carotenoids was performed. C. minutissima showed the best performance in shaker and was chosen for the tests in photobioreactor. The light intensity of 17,000 lx presented the best results. The temperature showed to have significant influence over biomass concentration, specific growth rate and CO2 biofixation rate, but the salt concentration only affected the specific growth rate. The temperature of 25 °C allowed the highest biomass productivity (0.094 g.L-1.d-1) and biomass concentration (0.43 g.L-1), independent of salt concentration, and the highest specific growth rate (0.81 d-1) and CO2 biofixation rate (12 gCO2.m-3.h-1) at the salt concentration of 37 g.L-1. The average lipid content of the microalgae was 13.2 % and the total carotenoids content were about 0.25 % of the cell dry weight at all temperatures and salt concentrations tested. It was possible to identify the production of the carotenoids lutein, zeaxanthin and β-carotene.

Neste trabalho é proposto o desenvolvimento de um processo para a biofixação do dióxido de carbono através do uso de microalgas. Para o cultivo desses microrganismos foram utilizados fotobiorreatores do tipo airlift. Nos cultivos foram avaliadas espécies de microalgas (Chlorella sp. e Chlorella minutissima), influência da intensidade luminosa (2.200 a 24.500 lx), concentração salina (28 a 40 g.L-1) e temperatura (25 °C a 35 °C) sobre a concentração de biomassa, velocidade específica de crescimento, produtividade de biomassa, biofixação de CO2, conteúdo lipídico e carotenoides totais. Também foi realizada a identificação dos carotenoides. A microalga escolhida para os testes em fotobiorreatores foi a C. minutissima. A intensidade luminosa que apresentou os melhores resultados foi a de 17.000 lx. A temperatura mostrou possuir influência significativa na concentração de biomassa, na velocidade específica de crescimento e na biofixação de carbono, mas a concentração salina influenciou apenas a velocidade específica de crescimento. A temperatura de 25 °C apresentou as maiores produtividade de biomassa (0,094 g.L-1.d-1) e concentração de biomassa (0,43 g.L-1), independente da concentração salina, e as maiores velocidade específica de crescimento (0,81 d-1) e biofixação de CO2 (12 gCO2.m-3.h-1), na concentração salina de 37 g.L-1. O conteúdo lipídico médio das microalgas foi de 13,2 % e os carotenoides totais apresentaram teor de 0,25 % do peso seco das microalgas em todas as condições de concentração salina e temperatura testadas. Foi possível identificar a produção dos carotenoides luteína, zeaxantina e β-caroteno. / The present study proposes the development of a process for carbon dioxide biofixation through the use of microalgae. Flat-plate airlift photobioreactors were used. Microalgaes species (Chlorella sp. and Chlorella minutissima) and the influence of light intensity (2,200 to 24,000 lx), salt concentration (28 to 40 g.L-1) and temperature (25 to 35 °C) over biomass concentration, specific growth rate, biomass productivity, CO2 biofixation rate, lipid content and total carotenoids content were evaluated. The identification of the carotenoids was performed. C. minutissima showed the best performance in shaker and was chosen for the tests in photobioreactor. The light intensity of 17,000 lx presented the best results. The temperature showed to have significant influence over biomass concentration, specific growth rate and CO2 biofixation rate, but the salt concentration only affected the specific growth rate. The temperature of 25 °C allowed the highest biomass productivity (0.094 g.L-1.d-1) and biomass concentration (0.43 g.L-1), independent of salt concentration, and the highest specific growth rate (0.81 d-1) and CO2 biofixation rate (12 gCO2.m-3.h-1) at the salt concentration of 37 g.L-1. The average lipid content of the microalgae was 13.2 % and the total carotenoids content were about 0.25 % of the cell dry weight at all temperatures and salt concentrations tested. It was possible to identify the production of the carotenoids lutein, zeaxanthin and β-carotene.