[Show abstract][Hide abstract] ABSTRACT: Digestibility of a micro-algal mixture was evaluated by mesophilic anaerobic digestion in continuously-stirred tank reactors. The culture consisted primarily of Scenedesmus spp. continuously cultivated over a 6-month period in a 100 m2 raceway reactor instrumented to record pH, dissolved oxygen and temperature. The raceway received supplementary carbon in the form of flue gas from a diesel boiler (10% CO2) injected into a 1-m deep sump to control pH in the range 7.8–8.0. Dilution was optimised to biomass productivity and gave values of 10–15 and 20–25 g total suspended solids (TSS) m− 2 day− 1 in winter (December–February) and spring (April–May), respectively. The culture for the anaerobic digestion trial was harvested in February by centrifugation to give an algal paste containing 4.3% volatile solids (VS). Semi-continuous digestion at organic loading rates of 2.00, 2.75 and 3.50 g VS l− 1 day− 1 gave volumetric biogas productions of ~ 0.66, ~ 0.83 and ~ 0.99 l l− 1 day− 1, respectively. Specific methane yield ranged from 0.13 to 0.14 l CH4 g− 1 VSadded with biogas methane content ~ 62%. Overall the digestion process was stable, but only ~ 30% VS destruction was achieved indicating low biodegradability, due to the short retention times and the recalcitrant nature of this type of biomass.
[Show abstract][Hide abstract] ABSTRACT: A process for the production of l-amino-acids concentrates from microalgae biomass by enzymatic hydrolysis has been developed. The process includes pre-treatment for cell-disruption, enzymatic hydrolysis and final separation by centrifugation. Thermal and mechanical cell-disruption methods have been tested, selecting mechanical disruption using bead milling for 30 min. The enzymatic hydrolysis was done using the commercial enzymes Alcalase and Flavourzyme. Maximum hydrolysis was obtained for biomass concentrations under 270 g/l and previous additional treatment with Viscozyme, reaching a 42% hydrolysis. Repeated reaction steps increased the hydrolysis from 42% (4h) with a single step to 59% (8h) after two successive steps. Further increase of the number of steps had a meagre impact on the global yield. The process widens the portfolio of products that can be obtained from microalgae biomass and is a new possibility to enhance the economic viability of microalgae-based biofuels production processes.
[Show abstract][Hide abstract] ABSTRACT: This paper focuses on modelling the growth rate and exopolysaccharides production of Anabaena sp. ATCC 33047, to be used in carbon dioxide removal and biofuels production. For this, the influence of dilution rate, irradiance and aeration rate on the biomass and exopolysaccharides productivity, as well as on the CO(2) fixation rate, have been studied. The productivity of the cultures was maximum at the highest irradiance and dilution rate assayed, resulting to 0.5 g(bio) l(-1) day(-1) and 0.2 g(eps) l(-1) day(-1), and the CO(2) fixation rate measured was 1.0 gCO(2) l(-1) day(-1). The results showed that although Anabaena sp. was partially photo-inhibited at irradiances higher than 1,300 μE m(-2) s(-1), its growth rate increases hyperbolically with the average irradiance inside the culture, and so does the specific exopolysaccharides production rate. The latter, on the other hand, decreases under high external irradiances, indicating that the exopolysaccharides metabolism hindered by photo-damage. Mathematical models that consider these phenomena have been proposed. Regarding aeration, the yield of the cultures decreased at rates over 0.5 v/v/min or when shear rates were higher than 60 s(-1), demonstrating the existence of thus existence of stress damage by aeration. The behaviour of the cultures has been verified outdoors in a pilot-scale airlift tubular photobioreactor. From this study it is concluded that Anabaena sp. is highly recommended to transform CO(2) into valuable products as has been proved capable of metabolizing carbon dioxide at rates of 1.2 gCO(2) l(-1) day(-1) outdoors. The adequacy of the proposed equations is demonstrated, resulting to a useful tool in the design and operation of photobioreactors using this strain.
Applied Microbiology and Biotechnology 02/2012; 94(3):613-24. · 3.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A new methodology to use efficiently flue gases as CO(2) source in the production of photosynthetic microorganisms is proposed. The CO(2) is absorbed in an aqueous phase that is then regenerated by microalgae. Carbonated solutions could absorb up to 80% of the CO(2) from diluted gas reaching total inorganic carbon (TIC) concentrations up to 2.0 g/L. The pH of the solution was maintained at 8.0-10.0 by the bicarbonate/carbonate buffer, so it is compatible with biological regeneration. The absorption process was modeled and the kinetic parameters were determined. Anabaena sp. demonstrated to tolerate pH (8.0-10.0) and TIC (up to 2.0 g/L) conditions imposed by the absorption step. Experiments of regeneration of the liquid phase demonstrated the feasibility of the overall process, converting CO(2) into organic matter. The developed process avoids heating to regenerate the liquid whereas maximizing the efficiency of CO(2) use, which is relevant to achieve the commercial production of biofuels from microalgae.
Biotechnology and Bioengineering 01/2012; 109(7):1637-50. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Maximum photobioreactor (PBR) efficiency is a must in applications such as the obtention of microalgae-derived fuels. Improving PBR performance requires a better understanding of the "light regime", the varying irradiance that microalgal cells moving in a dense culture are exposed to. We propose a definition of light regime that can be used consistently to describe the continuously varying light patterns in PBRs as well as in light/dark cycles. Equivalent continuous and light/dark regimes have been experimentally compared and the results show that continuous variations are not well represented by light/dark cycles, as had been widely accepted. It has been shown that a correct light regime allows obtaining photosynthetic rates higher than the corresponding to continuous light, the so-called "flashing light effect" and that this is possible in commercial PBRs. A correct PBR operation could result in photosynthetic efficiency close to the optimum eight quanta per O(2).
[Show abstract][Hide abstract] ABSTRACT: Lutein is an antioxidant that has gathered increasing attention due to its potential role in preventing or ameliorating age-related macular degeneration. Currently, it is produced from marigold oleoresin, but continuous reports of lutein-producing microalgae pose the question if those microorganisms can become an alternative source. Several microalgae have higher lutein contents than most marigold cultivars and have been shown to yield productivities hundreds of times higher than marigold crops on a per square meter basis. Microalgae and marigold are opposite alternatives in the use of resources such as land and labor and the prevalence of one or the other could change in the future as the lutein demand rises and if labor or land becomes more restricted or expensive in the producing countries. The potential of microalgae as a lutein source is analyzed and compared to marigold. It is suggested that, in the current state of the art, microalgae could compete with marigold even without counting on any of the improvements in microalgal technology that can be expected in the near future.
Applied Microbiology and Biotechnology 03/2010; 86(1):27-40. · 3.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Scenedesmus almeriensis is a fast-growing highly productive new strain and is also a good producer of lutein.The aim of this study was to determine the influences of pressure and temperature on the supercritical fluid extraction of lutein and β-carotene from a freeze-dried powder of the marine microalga, S. almeriensis. The operating conditions were as follows: pressure in the range 200–600bar and temperatures between 32 and 60°C. The extracts were analysed by HPLC. Empirical correlations were also developed.The results demonstrate that it is necessary to work at a pressure of 400bar and a temperature of 60°C to obtain a significant yield in the extraction of pigments. In comparison with the reference extraction process used, the results show that better yields are obtained in the extraction of β-carotene; it is possible to extract 50% of the total of this pigment contained in the microalga studied.
[Show abstract][Hide abstract] ABSTRACT: In this paper the utilization of the cyanobacteria Anabaena sp. in carbon dioxide removal processes is evaluated. For this, continuous cultures of this strain were performed at different dilution rates; alternatives for the recovery of the organic matter produced being also studied. A maximum CO(2) fixation rate of 1.45 g CO(2) L(-1) day(-1) was measured experimentally, but it can be increased up to 3.0 g CO(2) L(-1) day(-1) outdoors. The CO(2) is mainly transformed into exopolysaccharides, biomass representing one third of the total organic matter produced. Organic matter can be recovered by sedimentation with efficiencies higher than 90%, the velocity of sedimentation being 2.10(-4) s(-1). The major compounds were carbohydrates and proteins with productivities of 0.70 and 0.12 g L(-1) day(-1), respectively. The behaviour of the cultures of Anabaena sp. has been modelized, also the characteristics parameters requested to design separation units being reported. Finally, to valorizate the organic matter as biofertilizers and biofuels is proposed.
[Show abstract][Hide abstract] ABSTRACT: Bacterial growth on mixed substrates is employed for wastewater treatment. Biodegradation kinetics of Pseudomonas putida CECT 324 growth on formic acid, vanillin, phenol and oxalic acid mixtures is described. The experiments were carried out in a stirred-tank fermentor in batch mode at different temperatures (25, 30 and 35 degrees C) and pH (5, 6 and 7). The four compounds selected are typical intermediates in pesticide-contaminated water treated by advanced oxidation processes (AOPs). The toxicity of intermediates was investigated for a combined AOP-biological treatment, and the minimum DOC inhibitory concentration of the intermediate mixture was 175 ppm. The resulting biodegradation and growth kinetics were best described by the sum kinetics with interaction parameters (SKIP) model. Phenol and oxalic acid inhibit P. putida growth, and formic acid consumption strongly affects the biodegradation of oxalic acid. At all the temperatures tested and at pH between 5 and 7, P. putida CECT 324 was able to degrade the four substrates after culture times of 30 h at 30 degrees C and pH 7, which were the best conditions, and after 70 h, under the worst, at 35 degrees C.
Journal of Hazardous Materials 04/2008; 151(2-3):780-8. · 3.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Biodegradability of a partially photo-oxidized pesticide mixture is demonstrated and the effect of photo-Fenton treatment time on growth and substrate consumption of the bacteria Pseudomonas putida CECT 324 is shown. Four commercial pesticides, laition, metasystox, sevnol and ultracid, usually employed in citric orchards in eastern Spain, were chosen for these experiments. The active ingredients are, respectively, dimethoate, oxydemeton-methyl, carbaryl and methidathion. Judging by biomass measurements, dissolved organic carbon measurements and biodegradation efficiency, it may be concluded that 90min<t(30W)<110min is the critical point for the photo-Fenton treatment. P. putida is sensitive to photo-produced intermediates giving rise to different kinetic behaviour: longer lag phases, slower growth rates and lower carbon uptake rates. Nonetheless, the percentage of carbon consumption was over 80%, pointing out the biodegradability of the mixture. Biodegradation efficiencies (E(f)) of the photo-reaction intermediates were around 60%, in small 50-ml cultures and in a 12-l bubble column bioreactor. But with the main difference that E(f) in the former took 120h and the same biodegradation was reached in less than 30h in the latter. Therefore, for qualitative results, experiments in flasks might be recommendable, but not for quantitative results for designing purposes.
[Show abstract][Hide abstract] ABSTRACT: The present paper makes a comparative analysis of the outdoor culture of H. pluvialis in a tubular photobioreactor and a bubble column. Both reactors had the same volume and were operated in the same way, thus allowing the influence of the reactor design to be analyzed. Due to the large changes in cell morphology and biochemical composition of H. pluvialis during outdoor culture, a new, faster methodology has been developed for their evaluation. Characterisation of the cultures is carried out on a macroscopic scale using a colorimetric method that allows the simultaneous determination of biomass concentration, and the chlorophyll, carotenoid and astaxanthin content of the biomass. On the microscopic scale, a method was developed based on the computer analysis of digital microscopic images. This method allows the quantification of cell population, average cell size and population homogeneity. The accuracy of the methods was verified during the operation of outdoor photobioreactors on a pilot plant scale. Data from the reactors showed tubular reactors to be more suitable for the production of H. pluvialis biomass and/or astaxanthin, due to their higher light availability. In the tubular photobioreactor biomass concentrations of 7.0 g/L (d.wt.) were reached after 16 days, with an overall biomass productivity of 0.41 g/L day. In the bubble column photobioreactor, on the other hand, the maximum biomass concentration reached was 1.4 g/L, with an overall biomass productivity of 0.06 g/L day. The maximum daily biomass productivity, 0.55 g/L day, was reached in the tubular photobioreactor for an average irradiance inside the culture of 130 microE/m2s. In addition, the carotenoid content of biomass from tubular photobioreactor increased up to 2.0%d.wt., whereas that of the biomass from the bubble column remained roughly constant at values of 0.5%d.wt. It should be noted that in the tubular photobioreactor under conditions of nitrate saturation, there was an accumulation of carotenoids due to the high irradiance in this reactor, their content in the biomass increasing from 0.5 to 1.0%d.wt. However, carotenoid accumulation mainly took place when nitrate concentration in the medium was below 5.0mM, conditions which were only observed in the tubular photobioreactor. A similar behaviour was observed for astaxanthin, with maximum values of 1.1 and 0.2%d.wt. measured in the tubular and bubble column photobioreactors, respectively. From these data astaxanthin productivities of 4.4 and 0.12 mg/L day were calculated for the tubular and the bubble column photobioreactors. Accumulation of carotenoids was also accompanied by an increase in cell size from 20 to 35 microm, which was only observed in the tubular photobioreactors. Thus it may be concluded that the methodology developed in the present study allows the monitoring of H. pluvialis cultures characterized by fast variations of cell morphology and biochemical composition, especially in outdoor conditions, and that tubular photobioreactors are preferable to bubble columns for the production of biomass and/or astaxanthin.
Journal of Biotechnology 06/2006; 123(3):329-42. · 3.18 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The present study outlines a process for the cost-effective production of 13C/15N-labelled biomass of microalgae on a commercial scale. The core of the process is a bubble column photobioreactor with exhaust gas recirculation by means of a low-pressure compressor. To avoid accumulation of dissolved oxygen in the culture, the exhaust gas is bubbled through a sodium sulphite solution prior to its return to the reactor. The engineered system can be used for the production of 13C, 15N, and 13C-15N stable isotope-labelled biomass as required. To produce 13C-labelled biomass, 13CO2 is injected on demand for pH control and carbon supply, whereas for 15N-labelled biomass Na15NO3 is supplied as nitrogen source at the stochiometric concentration. The reactor is operated in semicontinuous mode at different biomass concentrations, yielding a maximum mean biomass productivity of 0.3 gL(-1) day(-1). In order to maximize the uptake efficiency of the labelled substrates, the inorganic carbon is recovered from the supernatant by acidification/desorption processes, while the nitrate is delivered at stochiometric concentration and the harvesting of biomass is performed when the 15NO3- is depleted. In these conditions, elemental analysis of both biomass and supernatant shows that 89.2% of the injected carbon is assimilated into the biomass and 6.9% remains in the supernatant. Based on elemental analysis, 97.8% of the supplied nitrogen is assimilated into the biomass and 1.3% remains in the supernatant. Stable isotope-labelling enrichment has been analysed by GC-MS results showing that the biomass is highly labelled. All the fatty acids are labelled; more than 96% of the carbon present in these fatty acids is 13C. The engineered system was stably operated for 3 months, producing over 160 g of 13C and/or 15N-labelled biomass. The engineered bioreactor can be applied for the labelling of various microalgae.
[Show abstract][Hide abstract] ABSTRACT: The influence of fluid-dynamic conditions on the yield of Phaeodactylum tricornutum microalgal cultures was analyzed in two stages: first, the influence of air flow rate; second, the influence of using fluid-moving pumps for recirculating the culture. With respect to the air flow rate, the yield of the cultures increased with the aeration rate up to values of 2.0 v/v/min, then stress was observed and the yield of the cultures decreased. With respect to the influence of mechanical power supply for liquid impulsion, three different types of pumps--centrifugal, pulse, and peristaltic--were essayed at different power supplies. The cultures were stressed for the three types of pumps essayed. For each pump, the higher the power supply the lower was the Fv/Fm value and the higher was the stress at which cells were exposed. The highest measured stress was when the culture was moved with the centrifugal pump. Despite measured stress, for all the experiments stable steady states were reached, thus indicating that cells reduced their yield but did not die, as was verified by cell viability measurements. It was observed that the increase of the power supply improved the frequency of light exposition thus enhancing the yield of the cultures. However, the higher the power supply, the lower the microeddy length scale; therefore, stress could appear. Data demonstrated that the microeddy length scale was always much higher than cell size and therefore the turbulence was not responsible for stress. Also, the mass transfer was discarded as responsible for yield reduction. It was concluded that the shear rate was the factor determining the existence of stress phenomena. The evaluation of these shear rates demonstrated that values above 30-80 s(-1) damaged the cells strongly. These data were verified in an outdoor pilot-scale tubular photobioreactor that was implemented with the same type of pumps, thus demonstrating the necessity to take into account this factor in the design and scale-up of microalgal photobioreactors.
Biotechnology and Bioengineering 10/2004; 87(6):723-33. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The optimization of carbon use in pilot-scale outdoor tubular photobioreactors is investigated in this study. The behavior of a 0.20-m(3) tubular photobioreactor was studied, with and without algae, by steady-state and pulse dynamic-response analysis experiments. A model of the system was obtained and implemented in a programmable control unit and was used to control the reactor under normal production conditions. Results showed that, using and on-off control, the mean daily CO(2) flow in the reactor was 0.86 g min(-1), 19.7% of this being lost. By using a predictive control algorithm the mean daily CO(2) flow was reduced to 0.74 g min(-1), with losses being reduced to 15.6%. In this case, pH tracking was not adequate, especially at the beginning and end of the daylight period, because the variation in solar irradiance was not considered. Taking solar irradiance into account resulted in better performance, with mean daily CO(2) flow reduced to 0.70 g min(-1), and carbon losses reduced to 5.5%. pH tracking was improved and valve actuation was reduced. Improvement of pH control reduced pH gradients in the culture, which increased the photosynthesis rate and biomass productivity of the system. Biomass productivity increased from 1.28 to 1.48 g L(-1) day-(1) when on-off control was replaced by model-based predictive control plus solar irradiance effect mode. Implementation of this methodology in outdoor photobioreactors can increase productivity by 15% and reduce the cost of producing biomass by >6%. Clearly, application of effective control techniques, such as model-based predictive control (MPC), must be considered when developing these processes.
Biotechnology and Bioengineering 12/2003; 84(5):533-43. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The production of microalga Phaeodactylum tricornutum in an outdoor helical reactor was analysed. The influence of temperature, solar irradiance and air flow rate on the yield of the culture was evaluated. Biomass productivities up to 1.5 g l(-1) per day and photosynthetic efficiency up to 14% were obtained by maintaining the cultures below 30 degrees C, dissolved oxygen levels less than 400% Sat. (with respect to air saturated culture) and controlling the cell density in order to achieve an average irradiance within the culture below 250 microE m(-2) s(-1). Under these conditions, the fluorescence parameter, Fv/Fm, which reflects the maximal efficiency of PSII photochemistry, remained roughly 0.6-0.7 and growth rates up to 0.050 h(-1) were achieved. The average irradiance and the light/dark cycle frequency, were the variables determining the behaviour of the cultures. A hyperbolic relationship between growth rate and biomass productivity with the average irradiance was observed, whereas both biomass productivity and photosynthetic efficiency linearly increased with the light/dark cycle frequencies. Optimum design and operational conditions which maximise the production of P. tricornutum biomass in outdoor helical reactors were determined.
Journal of Biotechnology 07/2003; 103(2):137-52. · 3.18 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The production of the microalga Phaeodactylum tricornutum in an outdoor helical reactor was analyzed. First, fluid dynamics, mass-transfer capability, and mixing of the reactor was evaluated at different superficial gas velocities. Performance of the reactor was controlled by power input per culture volume. A maximum liquid velocity of 0.32 m s(-1) and mass transfer coefficient of 0.006 s(-1) were measured at 3200 W m(-3). A model of the influence of superficial gas velocity on the following reactor parameters was proposed: gas hold-up, induced liquid velocity, and mass transfer coefficient, with the accuracy of the model being demonstrated. Second, the influence of superficial gas velocity on the yield of the culture was evaluated in discontinuous and continuous cultures. Mean daily values of culture parameters, including dissolved oxygen, biomass concentration, chlorophyll fluorescence (F(v)/F(m) ratio), growth rate, biomass productivity, and photosynthetic efficiency, were determined. Different growth curves were measured when the superficial gas velocity was modified-the higher the superficial gas velocity, the higher the yield of the system. In continuous mode, biomass productivity increased by 35%, from 1.02 to 1.38 g L(-1) d(-1), when the superficial gas velocity increased from 0.27 to 0.41 m s(-1). Maximal growth rates of 0.068 h(-1), biomass productivities up to 1.4 g L(-1) d(-1), and photosynthetic efficiency of up to 15% were obtained at the higher superficial gas velocity of 0.41 m s(-1). The fluorescence parameter, F(v)/F(m), which reflects the maximal efficiency of PSII photochemistry, showed that the cultures were stressed at average irradiances within the culture higher than 280 microE m(-2) s(-1) at every superficial gas velocity. For nonstressed cultures, the yield of the system was a function of average irradiance inside the culture, with the superficial gas velocity determining this relationship. When superficial gas velocity was increased, higher growth rates, biomass productivities, and photosynthetic efficiencies were obtained for similar average irradiance values. The higher the superficial gas velocity, the higher the liquid velocity, with this increase enhancing the movement of the cells inside the culture. In this way the efficiency of the cells increased and higher biomass concentrations and productivities were reached for the same solar irradiance.
Biotechnology and Bioengineering 05/2003; 82(1):62-73. · 4.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Commercial production of intracellular microalgal metabolites requires the following: (1) large-scale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study.