Microalgae have been widely reported as a promising source of biofuels, mainly based on their high areal productivity of biomass and lipids as triacylglycerides and the possibility for cultivation on non-arable land. The isolation and selection of suitable strains that are robust and display high growth and lipid accumulation rates is an important prerequisite for their successful cultivation as a bioenergy source, a process that can be compared to the initial selection and domestication of agricultural crops. We developed standard protocols for the isolation and cultivation for a range of marine and brackish microalgae. By comparing growth rates and lipid productivity, we assessed the potential of subtropical coastal and brackish microalgae for the production of biodiesel and other oil-based bioproducts. This study identified Nannochloropsis sp., Dunaniella salina and new isolates of Chlorella sp. and Tetraselmis sp. as suitable candidates for a multiple-product algae crop. We conclude that subtropical coastal microalgae display a variety of fatty acid profiles that offer a wide scope for several oil-based bioproducts, including biodiesel and omega-3 fatty acids. A biorefinery approach for microalgae would make economical production more feasible but challenges remain for efficient harvesting and extraction processes for some species.
Association of microalgae culture and anaerobic digestion seems a promising technology for sustainable algal biomass and biogas production. The use of digestates for sustaining the growth of microalgae reduces the costs and the environmental impacts associated with the substantial algal nutrient requirements. A natural marine algae-bacteria consortium was selected by growing on a medium containing macro nutrients (ammonia, phosphate and acetate) specific of a digestate, and was submitted to a factorial experimental design with different levels of temperature, light and pH. The microalgal consortium reached a maximum C conversion efficiency (i.e. ratio between carbon content produced and carbon supplied through light photosynthetic C conversion and acetate) of 3.6%. The presence of bacteria increased this maximum C conversion efficiency up to 6.3%. The associated bacterial community was considered beneficial to the total biomass production by recycling the carbon lost during photosynthesis and assimilating organic by-products from anaerobic digestion.
Nannochloropsis oculata was grown in an outdoor bubble column photobioreactor. To obtain information about the behaviour of microalgae/photobioreactor system related to the CO(2) net balance, an analysis of the pH profiles during microalgae growth was carried out. The use of the carbonate equilibrium chemistry and the overall CO(2) volumetric mass transfer in the photobioreactor has permitted to obtain information of the CO(2) losses/CO(2) microalgae consumption ratios. The simplicity of the technique used (a pH probe) could extend the use of this methodology for the correct selection of the photobioreactor/microalgae parameters with the aim to maximize the [CO(2) uptaken/(CO(2) uptaken+CO(2) stripped)] ratios.
In this paper subcritical co-solvents extraction (SCE) of algal lipid from wet pastes of Nannochloropsis sp. is examined. The influences of five operating parameters including the ratio between ethanol to hexane, the ratio of mixed solvents to algal biomass (dry weight), extraction temperature, pressure, and time were investigated. The determined optimum extraction conditions were 3:1 (hexane to ethanol ratio), 10:1 ratio (co-solvents to microalgae (dry weight) ratio), 90°C, 1.4 MPa, and 50 min, which could produce 88% recovery rate of the total lipids. In addition, electron micrographs of transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were conducted to show that the algal cell presented shrunken, collapsed with some wrinkles and microholes after SCE extraction. The main composition of total lipids extracted under the optimum conditions was TAG which represented more than 80%. And the fatty acid profile of triglycerides revealed that C16:0 (35.67 ± 0.2%), C18:1 (26.84 ± 0.044%) and C16:1 (25.96 ± 0.011%) were dominant.
Microalgae are microorganisms that can fix CO(2) by using the energy from the sun and transforming it into organic molecules such as lipids (i.e. feedstock for biodiesel production). Microfiltration is a promising method to be considered in the harvesting step. In this study, two antifouling methods were tested in order to minimize permeability decrease over time, at low trans-membrane pressure filtration. Preliminary experiments were performed to find optimum conditions of transmembrane pressure, rotational speed and membrane pore size. Pilot experiments were carried out in the optimal conditions using microalgae obtained from the culture step and from a previous concentration process based on sedimentation. Fouling was significantly minimized, and the permeability plateau increased up to 600L/h/m(2)/bar. Three microalgae species were tested: Phaeodactylum tricornutum (Pht), Nannochloropsis gaditana (Nng) and Chaetoceros calcitrans (Chc). An economic assessment was also performed, which demonstrated that dynamic filtration is economically more efficient than tangential cross-flow filtration.
In this work, the hydrodynamic characteristics in tubular photobioreactors with a series of helical static mixers built-in were numerically investigated using computational fluid dynamics (CFD). The influences of height and screw pitch of the helical static mixer and fluid inlet velocity on the cell trajectories, swirl numbers and energy consumption were examined. In order to verify the actual results for cultivation of microalgae, cultivation experiments of freshwater Chlorella sp. were carried out in photobioreactor with and without helical static mixer built-in at the same time. It was shown that with built-in helical static mixer, the mixing of fluid could be intensified, and the light/dark cycle could also be achieved which is of benefit for the growth of microalgae. The biomass productivity of Chlorella sp. in tubular photobioreactor with helical static mixer built-in was 37.26 % higher than that in the photobioreactor without helical static mixer.
The oleaginous microalga Chlorella sp. LAM-H screened from freshwater was proven to be a prospective feedstock for oil production according to its fatty acid composition. In order to enhance lipid production, response surface methodology (RSM) was used with central composite design (CCD) to optimize the heterotrophic cultivation of microalgae. The experiment results showed that a satisfactory second-order polynomial regression equation was achieved with a high coefficient of determination (R(2)=0.9911) in analysis of variance. The effects of individual factors and their interactions on lipid productivity were successfully revealed. The greatest lipid productivity reached 247.16mgl(-1)d(-1) under the optimal conditions of glucose concentration 26.2gl(-1), sodium nitrate concentration 2.06gl(-1) and temperature 28.18°C. Moreover, validation tests were performed and the results were very close to the predicted values. It was demonstrated that the obtained model was effective for predicting lipid productivity of the isolated microalga.
Microalgae are considered as an intriguing candidate for biofuel production due to their high biomass yield. Studies on bio-oil production through fast pyrolysis and upgrading to hydrocarbon fuels using algal biomass are limited as compared to other terrestrial biomass. Therefore, in this study, a fresh water green alga, Chlorella vulgaris, was taken for pyrolysis study. The average activation energy for pyrolysis zone was found to be 109.1kJ/mol. Fixed-bed pyrolysis of algae gave a bio-oil yield of 52.7wt.%, which accounts for 60.7wt.% carbon yield. In addition, analytical pyrolysis of C. vulgaris was carried out in a Py/GC-MS to identify major compounds present in bio-oil with and without catalyst (H(+)ZSM-5). The study found that in catalytic-pyrolysis, as the catalyst loading increased from zero to nine times of the biomass, the carbon yield of aromatic hydrocarbons increased from 0.9 to 25.8wt.%.
Microalgae may be the source of high amount of lipid and protein. It has the property for carbon dioxide sequestration, recycling and also can remove pollutants from wastewater. Using traditional methods, collection of algal biomass is either cost effective, time consuming or may be toxic due to use of chemical salts. The aim of this study is to harvest freshwater microalgae (Chlorella sp. CB4) biomass by using polymer. Polyacrylamide grafted starch (St-g-PAM) has been synthesized by microwave assisted method involving a synergism of microwave radiation and ceric ammonium nitrate (CAN) to initiate the grafting reaction. The synthesis was optimized in terms of CAN and monomer (acrylamide) concentration. The algal flocculation efficacy of all the grades of this graft copolymer was studied through standard 'Jar test' procedure. Effects of percentage grafting, pH and zeta potential on percentage recovery of algal biomass were thoroughly investigated.
For many years, benefits and disadvantages of pigments production either by microalgae or yeasts have been under analysis. In this contribution we shall deal with Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma) and Haematococcus pluvialis, which are known as major prominent microorganisms able to synthesize astaxanthin pigment. Then, the usual trend is to look for optimal conditions to conduct astaxanthin synthesis. From one side, pigment production by H. pluvialis is promoted under cellular stress conditions like nutrient deprivation, exposition to high light intensity, aeration. On the other side, X. dendrorhous is able to show significant increase in astaxanthin synthesis when grown in natural carbon sources like coconut milk, grape juice. The main aim of this chapter is to describe optimal environmental conditions for astaxanthin production by X. dendrorhous or H. pluvialis.
For overcoming the long period of seed cultured photoautotrophically and inadequate cell supply for the inoculation of microalgae photoautotrophic cultivation, a model for the photoautotrophic culture of three Chlorella species with heterotrophic cells as seed was investigated. The model can not only take advantages of rapid cell growth in heterotrophic process for preparation of cells as seed but also increase the biomass and lipid productivities of the microalgae cultivated photoautotrophically. The results showed that biomass productivities of Chlorella pyrenoidosa, Chlorella ellipsoidea and Chlorella vulgaris cultured by heterotrophy were 20.9, 26.9 and 25.2 times higher than those by photoautotrophy in seed culturing period. In the subsequent photoautotrophic culture, the biomass and lipid productivities of C. pyrenoidosa, C. ellipsoidea and C. vulgaris with heterotrophic seed were 1.91, 1.51, 1.48 and 1.66, 1.37, 1.42 times higher than those with photoautotrophic seed. Furthermore, the culture model was also carried out successfully outdoor.
A switchable hydrophilicity solvent (SHS) was studied for its effectiveness at extracting lipids from freeze-dried samples of Botryococcus braunii microalgae. The SHS N,N-dimethylcyclohexylamine extracted up to 22wt.% crude lipid relative to the freeze-dried cell weight. The solvent was removed from the extract with water saturated with carbon dioxide at atmospheric pressure and recovered from the water upon de-carbonation of the mixture. Liquid chromatography-mass spectrometry (LC-MS) showed that the extracted lipids contained high concentrations of long chain tri-, di- and mono-acylglycerols, no phospholipids, and only 4-8% of residual solvent. Unlike extractions with conventional organic solvents, this new method requires neither distillation nor the use of volatile, flammable or chlorinated organic solvents.
Microalgae are a source of renewable oil for liquid fuels. However, costs for dewatering/drying, extraction, and processing have limited commercial scale production of biodiesel from algal biomass. A wet lipid extraction procedure was developed that was capable of extracting 79% of transesterifiable lipids from wet algal biomass (84% moisture) via acid and base hydrolysis (90°C and ambient pressures), and 76% of those extracted lipids were isolated, by further processing, and converted to FAMEs. Furthermore, the procedure was capable of removing chlorophyll contamination of the algal lipid extract through precipitation. In addition, the procedure generated side streams that serve as feedstocks for microbial conversion to additional bioproducts. The capability of the procedure to extract lipids from wet algal biomass, to reduce/remove chlorophyll contamination, to potentially reduce organic solvent demand, and to generate feedstocks for high-value bioproducts presents opportunities to reduce costs of scaling up algal lipid extraction for biodiesel production.
The stringent regulations for discharging acid mine drainage (AMD) has led to increased attention on traditional or emerging treatment technologies to establish efficient and sustainable management for mine effluents. To assess new technologies, laboratory investigations on AMD treatment are necessary requiring a consistent supply of AMD with a stable composition, thus limiting environmental variability and uncertainty during controlled experiments. Additionally, biotreatment systems using live cells, particularly micro-algae, require appropriate nutrient availability. Synthetic AMD (Syn-AMD) meets these requirements. However, to date, most of the reported Syn-AMDs are composed of only a few selected heavy metals without considering the complexity of actual AMD. In this study, AMD was synthesised based on the typical AMD characteristics from a copper mine where biotreatment is being considered using indigenous AMD algal-microbes. Major cations (Ca, Na, Cu, Zn, Mg, Mn and Ni), trace metals (Al, Fe, Ag, Na, Co, Mo, Pb and Cr), essential nutrients (N, P and C) and high SO(4) were incorporated into the Syn-AMD. This paper presents the preparation of chemically complex Syn-AMD and the challenges associated with combining metal salts of varying solubility that is not restricted to one particular mine site. The general approach reported and the particular reagents used can produce alternative Syn-AMD with varying compositions. The successful growth of indigenous AMD algal-microbes in the Syn-AMD demonstrated its applicability as appropriate generic media for cultivation and maintenance of mining microorganisms for future biotreatment studies.
The use of microalgae in the skin care market is already established although the scientific rationale for their benefit was not clearly defined. In this work, the biological activities of dermatologic interest of the water extract from the microalga Botryococcus braunii (BBWE) were evaluated by a battery of in vitro assays. At concentrations ranging from 0.1 to 0.001 % (w/v) BBWE promoted adipocytes differentiation by inhibiting hormone-sensitive lipase, thus promoting triglyceride accumulation in the cells. BBWE also induced gene expression of proteins involved in the maintenance of skin cells water balance such as aquaporin-3 (AQP3), filaggrin (FLG) and involucrin (INV). 0.1 % BBWE increased the gene expression of AQP3 of 2.6-folds, that of FLG and INV of 1.5- and 1.9-folds, respectively. Moreover, it induced the biosynthesis of collagen I and collagen III by 80 and 40 %, respectively, compared to the untreated control. BBWE antioxidant activity, evaluated by oxygen radical absorbance capacity (ORAC) assay, was of 43.5 μmol Trolox per gram of extract: a quite high value among those found for other microalgae extracts. BBWE inhibited the inducible nitric oxide synthase (iNOS) gene expression and the consequent nitrite oxide (NO) production under oxidative stress. At a concentration of 0.02 % BBWE reduced by 50 % the expression of iNOS and by about 75 % the NO production. Taken together, the results demonstrated that B. braunii water extract exerted an array of biological activities concurring with the skin health maintenance; therefore, it is a potential bioactive ingredient to be included in cosmetic products.
In the past decade, H₂ production using the green microalga Chlamydomonas reinhardtii has been extensively studied under laboratory-scale photobioreactors, while information on outdoor cultures is still lacking. In this paper, the results of experiments conducted with sulfur-deprived cultures of C. reinhardtii carried out in a 50-L horizontal tubular photobioreactor are presented. Hydrogen production experiments were carried out under both artificial and direct solar light. In both cases, the H₂ output attained was 18-20% of what obtained in the laboratory. However, no significant changes in the H₂ production were observed when cells grown outdoors were tested under laboratory conditions. Chlorophyll fluorescence measurements showed that outdoor cultures were subjected to strong photo-inhibition, due to the combination of high solar light intensity and sulfur-deprivation. Indeed, H₂ production was only achieved outdoors when cultures were previously acclimated to sunlight, a condition that caused a number of physiological changes, namely: (i) a decrease in the chlorophyll content per unit of dry weight; (ii) an increase in the photosynthesis and respiration rates, and (iii) a higher induction of the xanthophyll cycle pigments as compared to non-acclimated cultures. It was concluded that the reduced H₂ output achieved in the 50-L photobioreactor was due to the different illumination pattern to which the cultures were exposed (one-sided vs. two-sided illumination provided in the laboratory), as well as to the great difference in the mixing times (60 min vs. 15.5s achieved in the lab-scale photobioreactor). To the very best of our knowledge this is the first time that H₂ production with green algae has been achieved by means of solar light.
A method was developed based on supercritical fluid chromatography for quantitative determination of carotenoids in extracts of Scenedesmus sp. By utilizing the low backpressure in supercritical fluid chromatography, a C18 column and a 2-ethyl pyridine column were coupled in series. It was concluded that even minor changes in temperature had a substantial effect on selectivity. A standard mixture of 8 carotenoids and microalgae extracts obtained through supercritical fluid extraction with and without 10% ethanol as a co-solvent were successfully separated. All of the carotenoids were separated within 10min, while the total analysis time was 20min. The method was validated and the carotenoids of microalgae extracts were quantified. Furthermore, the method should be seen as a more rapid and environmentally sustainable alternative to traditional high-performance liquid chromatography methods utilizing organic solvents.
The effect of ratio between autoflocculating and target microalgae in bio-flocculation was studied with emphasis on the recovery, sedimentation rate and energy demand for harvesting the target microalgae. When the autoflocculating microalgae Ettlia texensis, Ankistrodesmus falcatus and Scenedesmus obliquus were added to Chlorella vulgaris at a ratio of 0.25, the recovery of C. vulgaris increased from 25% to, respectively, 40%, 36% and 31%. The sedimentation rate increased as well. Addition of Tetraselmis suecica to Neochloris oleoabundans at a ratio of 0.25 increased the recovery from 40% to 50%. Application of bio-flocculation at a ratio of 0.25, followed by centrifugation reduces the energy demand for harvesting of the target microalgae from 13.8MJkgDW(-1) if only centrifugation is used to 1.83, 1.81, 1.53 and 1.34MJkgDW(-1), respectively, using T. suecica, E. texensis, A. falcatus and S. obliquus and 3h sedimentation before centrifugation.
The growth and lipid productivity of an isolated microalga Chlorella vulgaris ESP-31 were investigated under different media and cultivation conditions, including phototrophic growth (NaHCO(3) or CO(2), with light), heterotrophic growth (glucose, without light), photoheterotrophic growth (glucose, with light) and mixotrophic growth (glucose and CO(2), with light). C. vulgaris ESP-31 preferred to grow under phototrophic (CO(2)), photoheterotrophic and mixotrophic conditions on nitrogen-rich medium (i.e., Basal medium and Modified Bristol's medium), reaching a biomass concentration of 2-5 g/l. The growth on nitrogen-limiting MBL medium resulted in higher lipid accumulation (20-53%) but slower growth rate. Higher lipid content (40-53%) and higher lipid productivity (67-144 mg/l/d) were obtained under mixotrophic cultivation with all the culture media used. The fatty acid composition of the microalgal lipid comprises over 60-68% of saturated fatty acids (i.e., palmitic acid (C16:0), stearic acid (C18:0)) and monounsaturated acids (i.e., oleic acid (C18:1)). This lipid composition is suitable for biodiesel production.