Advances in Microalgal Research in Brazil

Abstract Microalgae research has attracted interest worldwide and in order to advance algal biotechnology in Brazil, government has been funding several projects. In the last 10 years, two main funds were provided by the National Council of Scientific and Technological Development (CNPq) agency to researchers in Brazil, who study the potential uses of microalgae for biomass, bioproducts and biofuels production. These funded projects addressed aspects of algal strain identification, development of algal cultivation techniques, designing photobioreactors and raceway ponds, modeling harvesting and dewatering process, maximizing biomass and oil productivities, characterizing chemical composition with different extractions systems and determining physiochemical properties of biodiesel. This review presents the state of art of algal research conducted by Brazilian institutions. Special attention is given to the recent progress on microalgal cultivation, high-value products extracted from microalgae and potential biofuels production. This review may serve as a policy instrument for planning next steps for algal research in Brazil as well as for attracting attention from international researchers who work with microalgae and would like to pursue a future partnership on algal research with Brazilian research institutions.

Crecimiento y metabolismo de Fischerella TB22 en medio de cultivo BG11 / Growth and metabolism of Fischerella TB22 in BG110 culture medium

Los microorganismos fijadores de nitrógeno de vida libre, abarcan una gama morfológica que va desde los organismos unicelulares como las bacterias y algunas cianobacterias, hasta multicelulares, filamentosas, por ello es importante conocer cómo se comportan y se puede saber haciendo una curva de crecimiento microbiano. Para este estudio se prepararon 4 fotobioreactores de columna burbujeada con inoculo de Fischerella TB22, se pusieron en aireación constante con 12 horas luz y 12 horas obscuridad durante 40 días con diferentes tratamientos de ajuste de volumen del medio de cultivo y ajuste del pH. El objetivo de este trabajo fue evaluar el crecimiento en biomasa por peso seco, densidad óptica, pH y amonio de Fischerella sp. en medio de cultivo BG110 durante 12 días. Las variables que se midieron de la curva de crecimiento de las cianobacterias, siguieron el patrón de una curva típica de crecimiento microbiano. (AU)

Free-living nitrogen-fixing microorganisms cover a morphological range that goes from unicellular organisms such as bacteria and some cyanobacteria, to multicellular, filamentous, therefore it is important to know how they behave and can be known by makinga microbial growth curve. For this study, 4 bubbled column photobioreactors with Fischerella TB22inoculum were prepared, they were placed in constant aeration with 12 hours of light and 12 hours of darkness for 40 days with different treatments of volumeadjustment of the culture medium and pH adjustment. The objective of this work was to evaluate the biomass growth by dry weight, optical density, pH, and ammonia of Fischerella sp. in the BG110 culture medium for 12 days. The variables that were measured from the growth curve of cyanobacteria followed the pattern of a typical microbial growth curve. (AU)

Kinetic characterization and modeling of a microalgae consortium isolated from landfill leachate under a high CO2 concentration in a bubble column photobioreactor

BACKGROUND: The determination of kinetic parameters and the development of mathematical models are of great interest to predict the growth of microalgae, the consumption of substrate and the design of photobioreactors focused on CO2 capture. However, most of the models in the literature have been developed for CO2 concentrations below 10%. RESULTS: A nonaxenic microalgal consortium was isolated from landfill leachate in order to study its kinetic behavior using a dynamic model. The model considered the CO2 mass transfer from the gas phase to the liquid phase and the effect of light intensity, assimilated nitrogen concentration, ammonium concentration and nitrate concentration. The proposed mathematical model was adjusted with 13 kinetic parameters and validated with a good fit obtained between experimental and simulated data. CONCLUSIONS: Good results were obtained, demonstrating the robustness of the proposed model. The assumption in the model of DIC inhibition in the ammonium and nitrate uptakes was correct, so this aspect should be considered when evaluating the kinetics with microalgae with high inlet CO2 concentrations.

Photocatalytic Degradation of Textile dye Orange-122 Via Electrospray Mass Spectrometry

Abstract This work reports the study of the potential application of Zn/TiO2 catalysts, obtained by the sol-gel method, in processes of environmental decontamination through the reactions of photodegradation of textile dye, followed by electrospray mass spectrometry. The catalysts synthesis was performed according to a 2² factorial design with repetition at the central point. The characterization techniques used were: N2 adsorption measurements (BET method), scanning electron microscopy with energy dispersive X-ray (MEV/EDS), X-ray diffraction and point of zero charge (PZC). The photocatalytic tests were performed in batch in the presence of sunlight, and to evaluate the degradation kinetics study, a rapid direct injection electrospray mass spectrometry (DI-ESI-MS) method has been developed. By the photocatalytic tests, the calcination temperature of 400 °C has shown the best results of discoloration for the reactive Orange-122 dye (99.76%) in a reaction time of 2h. The discoloration kinetics were a pseudo-first order, and a statistical analysis was performed to investigate the effects of the variables and to optimize the conditions of discoloration to the dye. After the reactional time of 2 h, an ion of m/z 441.5 was detected by ESI-MS, indicating that the photocatalytic process was effective for the degradation of the dye to secondary compounds.

Growth Kinetics of Nanofrustulum Shiloi Under Different Mixing Conditions in Flat-plate Photobioreactor

Abstract Diatoms are the major group of microalgae which have been utilized by the potential applications as food industries, aquatic feeds, cosmetics, biofuels, and pharmaceuticals. In this study, current approaches were made in order to determine growth rate, biomass productivity, protein, carbohydrate, lipid and fatty acid composition for Nanofrustulum shiloi cultures using both aeration and mixing conditions in flat-plate photobioreactor (PBR). Physical (the intensity of aeration, mixing, light intensity etc.) and chemical (nutritional materials) factors are affecting the growth and bioproduct contents of a diatom. Biomass and lipid productivities of N. shiloi were measured as 31.29 and 36.9622±0.0598 mg L-1 day-1 in flat-plate PBR having the combination of aeration and stirring system, respectively. A slightly higher amount of saturated fatty acids was detected in PBR having only bubbling system while the increase of mono- and poly- unsaturated fatty acids were found in PBR having the combination of aeration and stirring system. Flat-plate PBR design was also investigated for improving not only biomass but also the lipid productivity of N. shiloi.

Transformation of Brewery Subproducts into Valuable Biomass Using Mixotrophic Culture of Chlorella pyrenoidosa and Associated Bacteria

Abstract To develop a biorefinery concept applied in the brewery industry, Chlorella pyrenoidosa and a consortium of associated bacteria were cultivated mixotrophically in a continuous photobioreactor using brewery low-value subproducts as an integrative process. Beer production residues were biochemically characterized to assess the most promising options to be used as a nutrient source for microalgal cultivation. Due to its physical and chemical properties, pre-treated weak wort was used to prepare an organic complex culture medium for microalgal biotransformation. Filtration and nitrogen supplementation were necessary to improve nutrient removal and biomass productivity. Maximal removal of nitrate and phosphate obtained were 90% and 100% respectively. Depending on operation conditions, total carbohydrates depuration ranged from 50 - 80%. The initial concentration of total carbohydrates of the weak wort must be adjusted to 2 - 4g/L to maintain a stable equilibrium between microalgal and bacterial growth. The biochemical composition of produced biomass varied depending on the cultivation conditions as well as on its final use. Upon continuous mixotrophic conditions evaluated in this study, C. pyrenoidosa was composed mainly of carbohydrates and protein.

Processos de produção da proteína heteróloga mCherry por Chlamydomonas reinhardtii / Production processes of heterologous mCherry protein by Chlamydomonas reinhardtii

Este trabalho tem como finalidade estudar as melhores condições do cultivo da microalga Chlamydomonas reindhartii geneticamente modificada para a produção da proteína fluorescente mCherry a partir do estudo dos macronutrientes contidos no meio de cultivo. A proteína mCherry possui a vantagem de ser facilmente detectada no meio de cultivo por espectofotometria convencional, convertendo-se, desta forma, em uma molécula interessante para o estudo como modelo de expressão. Inicialmente, foram estudadas três diferentes fontes de nitrogênio para avaliar a expressão da proteína recombinante. Os resultados indicaram que a melhor fonte de nitrogênio para a produção da mCherry foi o NH4NO3. Em seguida, para avaliar os efeitos gerados pelos macronutrientes (acetato, cloreto de cálcio, sultato de magnésio, nitrato de amônio e fostato total) contidos no meio de cultivo TAP, foi realizado um planejamento composto central 25, em cultivos em microplacas, sendo os resultados avaliados por regressão multivariada. Além disso, a análise realizada por regressão multivariada indicou que, dos níveis avaliados das variáveis, as condições que melhor atendem à otimização da produção de mCherry e crescimento celular são: acetato, 33,35 mM; cloreto de cálcio, 0,45 mM; sulfato de magnésio, 0,83 mM; nitrato de amônio, 10,31 mM; fosfato total, 1,96 mM. Essas condições foram escolhidas para cultivo em fotobiorreator tubular, onde foi obtido título de fluorescência de mCherry a 608 nm de 59209 UF, correspondendo a um aumento de 118,5% maior que o título de fluorescência obtido com uso de meio TAP padrão. Com a finalidade de seguir com os processos de produção foi disenhado um biorreator tipo coluna e foi reaizado um estudio de produção em sistema semicontinuo. Os resultados obtidos demostraram que o sistema semicontinuo aumento 2,6 veces a produtividade da biomassa

This work aims to study the best conditions of the cultivation of the microalgae Chlamydomonas reindhartii genetically modified for the production of the fluorescent protein mCherry from the study of the macronutrients contained in the culture medium. The mCherry protein has the advantage of being easily detected in the culture medium by conventional spectrophotometry, thus becoming an interesting molecule for the study as an expression model. Initially, three different nitrogen sources were studied to evaluate the expression of the recombinant protein. The results indicated that the best source of nitrogen for the production of mCherry was NH4NO3. Then, to evaluate the effects generated by macronutrients (acetate, calcium chloride, magnesium sulphate, ammonium nitrate and total phosphate) contained in the TAP culture medium, a central composite 25 was carried out in cultures on microplates, Results evaluated by multivariate regression. In addition, multivariate regression analysis indicated that, from the evaluated levels of the variables, the conditions that best serve the optimization of mCherry production and cell growth are: acetate, 33.35 mM; Calcium chloride, 0.45 mM; Magnesium sulfate, 0.83 mM; 10.31 mM ammonium nitrate; Total phosphate, 1.96 mM. These conditions were chosen for cultivation in tubular photobioreactor where fluorescence titre of mCherry at 608 nm of 59209 UF was obtained, corresponding to an increase of 118.5% greater than the titer of fluorescence obtained using standard TAP medium. In order to follow the production processes, a column type bioreactor was designed and a production study was carried out in a semicontinuous system. The results showed that the semicontinuous system increased 2.6 times the productivity of the biomass

Produção de proteínas heterólogas em microalga / Production of heterologous protein in microalga

O objetivo desta tese foi explorar o sistema de produção de proteínas heterólogas em microalga com ênfase em Chlamydomonas reinhardtii por meio de: (1) desenvolvimento de um fotobiorreator tubular fechado de escala laboratorial, utilizando técnicas de manufatura digital; (2) avaliação de 7 diferentes proteínas fluorescentes (mTagBFP, Cerulean, Emerald, crGFP, cOFP, tdTomato e mCherry), como sistema reporter de secreção de proteínas em microalga; (3) avaliação do fotobiorreator desenvolvido utilizando cultivo de cepas recombinantes; (4) desenvolvimento de novos peptídeos sinais para secreção de proteínas em C. reinhardtii; (5) avaliação da produção de um biofármaco (hialuronidase) em microalgas, por meio da expressão de duas isoenzimas codificadas pelos genes HYA1 e SPAM1 em C. reinhardtii. O fotobiorreator tubular foi avaliado quanto a sua capacidade de resistir ao processo de esterilização por autoclavação e seu desempenho por meio do cultivo de cepa recombinante secretando mCherry. A fluorescência das proteínas fluorescentes foi medida por leitor de placas de fluorescência e visualizada intracelularmente por microscopia confocal de fluorescência. A atividade de hialuronidase foi determinada através de um ensaio enzimático turbidimétrico. O desenvolvimento do fotobiorreator resultou em um sistema fechado resistente a autoclavação, com capacidade de cultivo de cepas recombinantes de C. reinhardtii. Esse fotobiorreator proporcionou uma produtividade máxima de 10 mg/L.d de mCherry da cepa recombinante em sistema fechado, com velocidade específica de crescimento máxima de 1,27 d-1 para a cepa recombinante testada. Todas as proteínas fluorescentes avaliadas apresentaram capacidade de secreção por C. reinhardtii, com diferentes níveis de interferências em sua medição, permitindo a escolha da mCherry como proteína reporter. Entre os peptídeos sinais avaliados (quatro descritos na literatura - BiP, ARS1, CAH1 e IBP1 - e seis preditos), o peptídeo predito "SP5" foi o que apresentou maior capacidade de secreção, determinado por níveis de fluorescência no sobrenadante. A avaliação dos peptídeos sinais constatou a necessidade de explorar o desenvolvimento de sistemas de expressão (e.g. vetores de expressão) aliados a análises computacionais, como o SignalP 4.0. Por último, os dados desse estudo mostram que C. reinhardtii transformadas com o vetor de expressão foi capaz de produzir as duas isoformas de hialuronidase em sua forma ativa, evidenciando a capacidade desse sistema para a produção de biofármacos. Portanto, nesta tese o sistema de expressão de proteínas heterólogas baseado em microalgas foi explorado, atingindo os objetivos propostos. O fotobiorreator desenvolvido tem a capacidade de esterilização em escala laboratorial (1) e em cultivo com cepa recombinante propiciou elevada produtividade (3). As proteínas vermelhas fluorescentes apresentaram-se como as proteínas com menores interferências para estudos de secreção em C. reinhardtii (2). Além disso, o peptídeo predito SP5 apresentou o melhor desempenho na secreção de proteínas (4) e o vetor de expressão empregado permitiu a identificação de cepas produtoras de biofármaco hialuronidase (5). Portanto, o sistema de produção de proteínas heterólogas por microalgas é um sistema promissor e poderá permitir, utilizando sistemas de secreção, obter proteínas de alto valor comercial a baixos custos, empregando a secreção e técnicas de cultivo como a fermentação extrativa

In this thesis, the heterologous protein production in microalgae with emphasis on Chlamydomonas reinhardtii was explored through: (1) development of a laboratory scale closed tubular photobioreactor using digital manufacturing techniques; (2) evaluation of different fluorescent proteins (mTagBFP, Cerulean, Emerald, crGFP, cOFP, tdTomato and mCherry) as a reporter system for protein secretion in microalgae (3) evaluation of photobioreactor developed using recombinant strains culture; (4) development of new signals peptides for protein secretion in C. reinhardtii (5) expression evaluation of a biopharmaceutical (Hyaluronidase) in microalgae, through the expression of two isoenzymes encoded by the HYA1 and SPAM1 genes in C. reinhardtii. The tubular photobioreactor was evaluated for its ability to resist sterilization process by autoclaving and its performance by culturing recombinant strain secreting mCherry. Fluorescence of fluorescent proteins was measured by fluorescence plate reader and observed intracellularly by confocal fluorescence microscopy. The hyaluronidase activity was determined by a turbidimetric enzymatic assay. The development of the photobioreactor resulted in a closed system resistant to autoclaving, capable of culturing recombinant strains of C. reinhardtii. This recombinant strain achieved a maximum productivity of 10 mg/L.day of mcherry in the closed system, with a maximum growth rate of 1.27 d-1 for the recombinant strain tested. All the fluorescent proteins evaluated had C. reinhardtii secretion capacity, with different interference levels in their measurement, allowing the selection of mCherry as a reporter protein. Among the evaluated peptides (four described in the literature - BiP, ARS1, CAH1 and IBP1 - and six predicted), the predicted peptide "SP5" was the one that presented greater capacity of secretion, determined by levels of fluorescence in the supernatant. The results of this study point out the need to explore the development of biological systems (i.e., expression vectors) allied to computational analysis. Finally, the data from this study showed that C. reinhardtii could produce the two isoforms of hyaluronidase in its active form, evidencing the capacity of this system to produce biopharmaceuticals. Therefore, in this thesis the heterologous protein expression system based on microalgae was explored, reaching the proposed objectives. The developed photobioreator has sterilization capabilityin laboratorial scale (1) and in culture with recombinant strain had high productivity (3). The red fluorescent proteins was found as the most suitable proteins for studies of secretion in C. reinhardtii with lower interference levels(2). In addition, the predicted SP5 peptide showed the best performance in protein secretion (4) and the expression vector employed allowed the identification of strains producing biopharmaceutical hyaluronidase (5). Therefore, the system of heterologous proteins production by microalgae is promising and will allow, using secretion systems, to obtain proteins of high commercial value at low costs, using secretion and cultivation techniques such as extractive fermentation

Microalgae population dynamics in photobioreactors with secondary sewage effluent as culture medium

Nitrogen and phosphorus present in sewage can be used for microalgae growth, possibiliting cost reduction in the production of microalgae at the same time that it decreases the eutrophication potential of the effluent. This research aimed at monitoring the native community of microalgae and coliform bacteria in a secondary effluent from anaerobic municipal sewage treatment. Two treatments (aerated and non-aerated) were performed to grow microalgae under semi-controlled conditions in semi-closed photobioreactors in a greenhouse. The results showed no significant pH and coliforms (total and Escherichia coli) variation between treatments. Nutrient concentrations were reduced supporting microalgae growth up to 107 cells.mL−1 independent of aeration. Exponential growth was obtained from the first day for the non-aerated, but a 5 day lag phase of growth was obtained for the aerated. Chlorella vulgaris was the dominant microalgae (99.9%) in both treatments. In the aerated, 5 algae classes were detected (Chlorophyceae, Cyanophyceae, Chrysophyceae, Bacillariophyceae and Euglenophyceae), with 12 taxa, whereas in the non-aerated, 2 classes were identified (Chlorophyceae and Cyanophyceae), with 5 taxa. We concluded that effluent is viable for microalgae growth, especially Chlorella vulgaris, at the same time that the eutrophication potential and coliforms are decreased, contributing for better quality of the final effluent.

A novel flat plate photobioreactor for microalgae cultivation / 生物工程学报

Flashing light effect on microalgae could significantly improve the light efficiency and biomass productivity of microalgae. In this paper, the baffles were introduced into the traditional flat plate photobioreactor so as to enhance the flashing light effect of microalgae. Making Chlorella sp. as the model microalgae, the effect of light intensity and inlet velocity on the biomass concentration of Chlorella sp. and light efficiency were evaluated. The results showed that, when the inlet velocity was 0.16 m/s, with the increase of light intensity, the cell dry weight of Chlorella sp. increased and light efficiency decreased. With increasing the inlet velocity, the cell dry weight of Chlorella sp. and light efficiency both increased under the condition of 500 μmol/(m2 x s) light intensity. The cell dry weight of Chlorella sp. cultivated in the novel flat plate photobioreactor was 39.23% higher than that of the traditional one, which showed that the flashing light effect of microalgae could be improved in the flat plate photobioreactor with inclined baffles built-in.

Estudo da produção de biomassa e lipídios no cultivo de Neochloris oleoabundans sob diferentes condições de estresse nutricional e físico / Investigation on biomass and lipids accumulation in Neochloris oleoabundans cultivation under different nutritional and phisical stress conditions

As microalgas são candidatas promissoras para a produção em larga escala de biocombustíveis devido a sua alta eficiência fotossintética. No entanto, os custos relativamente altos de produção por baixas produtividades em lipídios têm sido um dos principais obstáculos que impedem sua produção comercial. Portanto, é necessário focar a pesquisa no aumento da biomassa e na produtividade em lipídios, através do desenvolvimento de biorreatores e técnicas de cultivo inovadoras. Numa primeira fase, este estudo mostra a otimização dos regimes de adição de nutrientes no cultivo de Neochloris oleoabundans em fotobiorreatores tubulares, determinando que a melhor metodologia de adição de CO2 é adicionando-o de forma intermitente e automatizada, enquanto que o melhor processo de alimentação de nitrogênio é por meio de um processo em batelada alimentada tomando como uma referência a produtividade diária de biomassa. Na segunda etapa, foi testada a influência de agentes estressores adicionados ao cultivo sob carência de nitrogênio, tais como tiossulfato de sódio como agente redutor e cloreto de sódio e glicerina como agentes de choque osmótico, buscando um acúmulo de lipídios na biomassa. Os resultados mostraram que o tiossulfato de sódio em 1,2 mM e o cloreto de sódio em 2,2 mM aumentaram o total de lipídios em 21% e 25%, respectivamente. Finalmente, foram testados diferentes regimes de luz, com um esquema 12:12, sendo 12 horas de luz fluorescente e 12 horas com um sistema distinto: escuro, diodos emissores de luz (LED) vermelha e LED branca. Os melhores resultados foram obtidos com LED branca, com um acúmulo de lipídios de até 27% da biomassa seca e uma concentração final de células de 2335mg/L, estabelecendo assim um método de iluminação econômica com alta produtividade (145mg / L dia)

Microalgae are promising candidates for large-scale global biofuel production because of their high photosynthetic efficiency. However, relatively high production costs due to low lipid productivity have been one of the major obstacles impeding their commercial production. Therefore, it is necessary to accurate the research into an increase in biomass and oil productivity, by means of novel bioreactors' design and cultivation techniques. On a first stage, this study shows the optimization of nutrients' addition regimes in Neochloris oleoabundans cultivation in tubular photobioreactors, finding that the best CO2 addition methodology is an automatized intermittent adding and the best feeding process for nitrogen is a fed-batch process taking as a reference the daily biomass productivity. On the second step, it was tested the influence of stressing agents added to the culture under nitrogen starvation, such as sodium thiosulphate for reducing environment and sodium chloride and glycerol for osmotic shock, aiming lipid accumulation in the biomass. The results showed that sodium thiosulphate at 1,2mM and sodium chloride at 2,2mM raised the total lipids up to 21% and 25% respectively. Finally, there were tested different light regimes, with a scheme 12:12, being 12 hours of fluorescent light and 12 hours of a singular system: dark, red light-emitting-diodes (LED) and white LED. The best results were obtained with white LED, with an accumulation up to 27% of dry biomass and a final cell concentration up to 2335mg/L, establishing an economic illumination method with high productivity

Optimization of photoautotrophic lipid production of Chlorella ellipsoidea seeded with heterotrophic cells / 生物工程学报

Photoautotrophic cultivation with heterotrophic cells as seeds (heterotrophic cells/photoautotrophic cultivation) is an effective way for the development of microalgal biofuel, but its development potential from the point of process optimization has not been investigated in literatures. To evaluate this, the optimizations of medium and culture conditions for Chlorella ellipsoidea were studied. In the heterotrophic stage, the biomass concentration reached 11.04 g/L with the optimized medium in flask, which were 28.0% higher than that with the original medium, and the biomass concentration reached 73.89 g/L in 5-L fermenter. In the photoautotrophic stage, the culture medium and conditions were studied in a 2-L column photobioreactor. The maximum biomass concentration, lipid content and lipid productivity reached 1.62 g/L, 36.34% and 6.1 mg/(L·h) under the optimal photoautotrophic conditions. The lipids were mainly composed of C16-C18 fatty acids, which were raw material suitable for biodiesel. After optimization, heterotrophic cells/photoautotrophic cultivation can significantly improve the capacity of biofuel production by Chlorella ellipsoidea, this method is also expected to be an efficient way for the cultivation of other microalgae that can grow heterotrophically.

Oxygen evolution characteristics of Spirulina platensis under various light conditions / 生物工程学报

The knowledge of oxygen evolution characteristics, which is a symbol of photosynthetic activity, under various light conditions is important for photobioreactor design and operation. In this study, we constructed a device to investigate oxygen evolution characteristics of Spirulina platensis under two different light regimes: 1) continuous illumination of various light intensities (14-6 500 micromol/(m2 x s)); 2) medium frequency L/D cycles of four different light intensities (69, 505, 1 330, 4 265 micromol/(m2s)). Light limited region, intermediate region, light saturated region and light inhibited region of light intensity were recognized according to their relationship with oxygen evolution rate (OER) under continuous illumination. Investigation of S. platensis under L/D cycles showed whether photosynthetic efficiency could be increased with increasing L/D frequency largely depended on the light intensity applied. The higher the light intensity, the larger the photosynthetic enhancement could be expected with the increase of L/D frequency. The largest light integration effect was found under L/D cycles of high light intensity (4 265 micromol/(m2 x s)) and medium light fraction (k = 0.6), while light integration effect was totally absent under low light fractions (k < 0.2). We also discussed their implications to the practical aspects of microalgae cultivation.

Progress in microalgae culture system for biodiesel combined with reducing carbon dioxide emission / 生物工程学报

Wastewater resources, CO2 emission reduction and microalgae biodiesel are considered as current frontier fields of energy and environmental researches. In this paper, we reviewed the progress in system of microalgae culture for biodiesel production by wastewater and stack gas. Multiple factors including microalgal species, nutrition, culture methods and photobioreactor, which were crucial to the cultivation of microalgae for biodiesel production, were discussed in detail. A valuable culture system of microalgae for biodiesel production or other high value products combined with the treatment of wastewater by microalgae was put forward through the optimizations of algal species and culture technology. The culture system coupled with the treatment of wastewater, the reduction of CO2 emission with the cultivation of microalgae for biodiesel production will reduce the production cost of microalgal biofuel production and the treatment cost of wastewater simultaneously. Therefore, it would be a promising technology with important environmental value, social value and economic value to combine the treatment of wastewater with the cultivation of microalgae for biodiesel production.

Ethanol production with starch-based Tetraselmis subcordiformis grown with CO2 produced during ethanol fermentation / 生物工程学报

A system coupling ethanol fermentation with microalgae culture was developed, in which CO2 produced during ethanol fermentation was used as carbon source for the growth of Tetraselmis subcordiformis, a microalgae accumulating starch intracellularly. The biomass concentration about 2.0 g DCW/L was achieved within the photobioreactor for the batch culture of 7 days, and intracellular starch accumulation was about 45%. Furthermore, ultrasonic pretreatment and enzymatic hydrolysis were applied to the microalgae biomass, and 71.1% of the intracellular starch was converted into glucose that was fermented sequentially to ethanol by Saccharomyces cerevisiae with an ethanol yield of 87.6% of the theoretical value, indicating that the microalgae biomass could be an alternative feedstock for ethanol production to save grain consumption, and in the meantime mitigate the CO2 emission.

Cultivating an oleaginous microalgae with municipal wastewater / 生物工程学报

Municipal wastewater is usually problematic for the environment. The process of oleaginous microalgal culture requires large amounts of nutrients and water. Therefore, we studied the feasibility of oleaginous microalgal culture of Scenedesmus dimorphus in bubbled column photobioreactor with municipal wastewater added with different nutrients. S. dimorphus could adapt municipal nutrient-rich wastewater by adding some nutrients as nitrogen, phosphorus, ferric ammonium citrate and trace elements, and the amounts of such nutrients have significant effects on cell growth, biomass yield and lipid accumulation. At optimum compositions of wastewater medium, the algal cell concentration could reach 8.0 g/L, higher than that of 5.0 g/L in standard BG11. Furthermore, S. dimorphus had strong capacity to absorb inorganic nitrogen and phosphorus from its culture water. There was almost no total nitrogen and phosphorus residues in culture medium after three or four days culturing when the adding mounts of nitrate and phosphate in wastewater medium were no more than 185.2 mg/L and 16.1 mg/L respectively under the experimental conditions. As a conclusion, it was feasible to cultivate oleaginous microalgae with municipal nutrient-rich wastewater, not only producing feedstock for algal biodiesel, but also removing inorganic nitrogen and phosphorus from wastewater.