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Journal of CO2 Utilization

journal homepage: www.elsevier.com/locate/jcou

From oil renery to microalgal biorenery

Eduardo Jacob-Lopes a,*, Telma Teixeira Franco b,1
a b

Food Science and Technology Department, Federal University of Santa Maria, UFSM, Av. Roraima 1000, 97105-900 Santa Maria, RS, Brazil School of Chemical Engineering, University of Campinas, UNICAMP, P.O. Box 6066, 13083-970 Campinas, SP, Brazil

A R T I C L E I N F O

A B S T R A C T

Article history: Received 7 December 2012 Received in revised form 7 June 2013 Accepted 8 June 2013 Keywords: Microalgae/cyanobacteria Photobioreactor Microalgal biorenery Carbon dioxide sequestration Biodiesel Carbon balance

The objective of this study was to develop an integrated system of biotransformation of carbon dioxide in oil reneries. The liquid and gaseous wastes from oil rening were considered for the cultivation of geli in a bubble column photobioreactor. Growth kinetics, carbon dioxide Aphanothece microscopica Na removal and oxygen release rates, carbon balance, lipid production and biofuel quality were determined. The results showed the potential use of oil renery wastes in microalgae-based systems under optimized culture conditions. The maximum specic growth rate was 1.4 day1 and the maximum carbon dioxide elimination capacity was 22.9 mg/L min. Each CO2 mass unit bioconverted resulted in approximately 0.75 O2 of mass units released. Carbon balance analyses indicated that a small fraction (3.64%) of the carbon dioxide is xed into a biomass form. Volatile organic compounds (92.0%) were the main products of carbon dioxide transformation in the photobioreactor, in such conditions. For the production of biodiesel, it is possible to obtain 0.08 glipid/L day with this process. The quality properties of the biodiesel were an ester content of 99.7%, a cetane number of 51.3, an iodine value of 79.9 gI2/100 g, a degree of insaturation of 65.3% and a cold lter plugging point of 24.9 8C. Based on these results, the process developed could be considered a promising emerging biorenery platform of waste-material-utilization type. 2013 Published by Elsevier Ltd.

1. Introduction Microalgae systems for carbon dioxide sequestration and production of chemicals are an emergent area, representing a great promise for industrial application. Several processes have demonstrated their capabilities mostly for food and feed industries, producing pigments and additives, as also for the industries of cosmetics. Although the initial research was developed more than 50 years ago [1], the systems have reached maturity only in the last twenty years, for full-scale industrial application of highvalue molecules but nor for commodities nor biofuel production [2,3]. Microalgae are described as desirable source of renewable energy, which can be associated to carbon dioxide mitigation and to less aggressive use of non-agricultural land (a fraction of the area required by conventional oil crops). Industrial ventures and academic groups reported that the main hurdles to overcome are related to light provision, economical culture media, carbon dioxide sources, reactor conguration and cell harvesting [4,5].

As the cost of culture media is a limiting factor in the commercialization of microalgae biomass production and the demand for water is an important drawback in biofuels production, its desirable that liquid industrial efuents are used for microalgae systems and also, that industrial ue gases can be integrated to it [6,7]. The purity and high temperature of a stationary industrial emission can prevent biological processes to occur, and they are associated with the presence of other gases that inhibit cellular growth [8,9]. This present work highlights a sustainable way of associating wastewater and a gaseous efuent of an oil renery to grow microalgae. An industrial gas stream, rich in CO2, was investigated for using it in photosynthesis. Therefore, the aim of this study was to develop an integrated system for biotransformation of carbon dioxide in oil reneries. The focus was on the measurement of growth kinetics, carbon dioxide removal and oxygen release rates, carbon balance, biodiesel production and biofuel quality. 2. Materials and methods 2.1. Microorganism and culture conditions

* Corresponding author. Tel.: +55 55 3220 8822. E-mail addresses: jacoblopes@pq.cnpq.br (E. Jacob-Lopes), franco@feq.unicamp.br (T.T. Franco). 1 Tel.: +55 1935212089; fax: +55 1935212089. 2212-9820/$ see front matter 2013 Published by Elsevier Ltd. http://dx.doi.org/10.1016/j.jcou.2013.06.001

geli (RSMan92) Axenic cultures of Aphanothece microscopica Na were originally isolated from the Patos Lagoon estuary, located in the state of Rio Grande do Sul, Brazil (328010 S528050 W). Stock

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2 E. Jacob-Lopes, T.T. Franco / Journal of CO2 Utilization xxx (2013) xxxxxx Table 1 Composition of wastewater from renery industry. Parameter pH Temperature (8C) TSS (mg/L) BOD (mg/L) Oil and grease (mg/L) Nitrite (mg/L) Nitrate (mg/L) Ammonia (mg/L) Phosphate (mg/L) Phenol (mg/L) Cyanide (mg/L) Values are means SD of all months considered. Value 8.3 0.24 28.1 2.41 0.13 0.00 14.0 1.36 4.6 0.38 0.1 0.00 15.4 0.32 1.2 0.10 0.5 0.00 0.02 0.00 0.04 0.00

cultures were propagated and maintained in synthetic BGN medium [10] with the following composition (g/L): K2HPO43H2O 3H2O (0.040), MgSO47H2O (0.075), EDTA (0.001), H3BO3 (2.860), MnCl24H2O (1.810), ZnSO47H2O (0.222), Na2MoO42H2O (0.390), CuSO45H2O (0.079), CaCl26H2O (0.040), NaNO3 (150), C6H8O7H2O (0.006), ammonium iron citrate (0.006) and pH 8.0. The incubation conditions used were 25 8C, photon ux density of 15 mmol m2 s1 and a photoperiod of 12 h in a growth chamber. For the experiments, the inoculum was acclimatized to the simulated industrial emission (5.0% of CO2, 0.9% of CO, 2.2% of H2, 1.8% of CH4, 18.7% of O2 and 70.7% of N2) by maintaining it for 30 days under gradual addition of the air contaminated with industrial gas. 2.2. Culture medium Wastewater from renery oil industry (Petrobras Inc., Paulinia, SP, Brazil) was collected from the discharge point of the activated sludge treatment for 8 months, from May to December of 2007 and pH, temperature, biochemical oxygen demand (BOD), nitrite, nitrate, ammonia, phosphate, phenol, cyanide, oil and grease, and total suspended solids (TSS) were analyzed following Standard Methods for Examination of Water and Wastewater [11] (Table 1). The efuent was supplemented with 25% of salts of the BGN medium. The composition used was dened in previous studies that aimed to reduce nutrients and water use [7]. 2.3. Gaseous streams An inventory of the emissions from renery oil industry (Petrobras Inc., Paulinia, SP, Brazil) was considered, aiming to identify carbon dioxide sources suitable to use in photobioreactors. Carbon dioxide loading, purity and temperature of airstream were the selection criteria. The selected gas emission was simulated in a laboratory through a primary standard gas mixture (accuracy of 1%) (Praxair, Inc., Brazil).

2.4. Photobioreactor Measurements were made in a bubble column photobioreactor (Fig. 1). The system was built in 4 mm thick glass, had an internal diameter of 70 mm, a height of 700 mm, and a nominal working volume of 2.0 L. The dispersion system for the reactor consisted of a porous stone diffuser with 1.5 cm of diameter (average orice diameter of 100 mm), located in the center of the column. The reactor was continuously illuminated with sixteen 20 W uorescent daylight-type tubes (Osram Sylvania, Brazil) connected in parallels, located in a photoperiod chamber. The duration of light cycles was controlled by a timer. Airow into the photobioreactor was provided via ltered air and synthetic industrial gas cylinders through Teon tubing. 2.5. Obtaining the kinetic data The experiments were carried out in photobioreactors operating in intermittent regime, fed with 2.0 L of culture medium. The experimental conditions were as follows: initial cell concentration

Fig. 1. Photobioreactor diagram. 1: Photoperiod chamber; 2: pH, temperature and CO2 analyser; 3: gas exit sampler; 4: pH, temperature and CO2 sensor; 5: photobioreactor; 6: gas diffuser; 7: system controlling the ow rate and mixture of the gases; 8: gas entrance sampler; 9: air ow meter; 10: CO2 ow meter. All dimensions in mm.

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of 0.1 g L1, isothermal reactor operating at a temperature of 30 8C, photon ux density of 150 mmol m2 s1, and continuous aeration of 1 VVM [12]. The simulated industrial gas stream was diluted with air to obtain CO2 concentrations of 5, 15 and 25% (v/v). The light cycle evaluated was 24:0 (day:night), respectively. The cell density, carbon dioxide, oxygen, inorganic carbon and total carbon concentrations were monitored every 12 h during the growth phase of the microorganism. The tests were carried out in duplicate and the kinetic data referred to the mean of four repetitions. 2.6. Kinetic parameters Biomass data were used to calculate the biomass productivity [PX = (Xi Xi1)(ti ti1)1, g/L day], the maximum specic growth rate [ln(Xi/X0) = mmaxt, day1] and the lipid productivity [PL = PXLC, g/L day], in which X0 is the initial biomass concentration, Xi is the biomass concentration at the time ti and Xi1 is the biomass concentration at the time ti1, t is the residence time and LC is the lipid content. Carbon dioxide concentration data were used to calculate the elimination capacity [EC = (Co Ci)QVR1, mg/L min] and removal efciency [RE = ((Ci Co)/Ci) 100, %], where Co and Ci correspond to the inlet and outlet CO2 concentration, respectively, Q is the gas ow, and VR is the reactor volume. 2.7. Analytical techniques 2.7.1. Operational control of the photobioreactor The photon ux density was adjusted and controlled by using a digital photometer (Spectronics, model XRP3000), measuring the light incident on the external reactor surface (accuracy of 4%). The temperature was controlled using thermostats (accuracy of 3%) and measured using a polarographic probe. The ow rates of the carbon dioxide, air and CO2 enriched air were determined using rotameters (AFSG 100 Key Instruments, accuracy of 5%). 2.7.2. Determination of cellular concentration Cell concentration was gravimetrically determined by ltering a known volume of culture medium through a 0.45 mm lter and drying at a temperature of 60 8C for a period of 24 h [11] (accuracy of 10%). 2.7.3. Characterization of the industrial gas Gas chromatography (GC) was used for the characterization of the industrial gas. Two gas chromatographs (GC-7A and GC-16A, Shimadzu, Kyoto, Japan) equipped with thermal conductivity detectors (accuracy of 1%) are used. Helium was used as the carrier gas. The separation of gases was carried out by three columns, silica gel 30/60, molecular sieve 5A 30/60 and molecular sieve 5A 80/100, at temperatures of 25, 25 and 72 8C, respectively. The gaseous samples were collected into evacuated vials. The vials were prepared before sampling using a sealing-chamber. Two milliliters of sample were injected, using a gas-tight syringe. The areas obtained using the integrator were compared with reference curves to determine the CO2, CO, CH4, O2, H2 and N2 concentrations. 2.7.4. Determinations of CO2 and O2 concentration proles CO2/O2 exchange rates in the photobioreactor were monitored o Paulo, Brazil), equipped with a column by GC (Cromacon, Sa Porapack Q80/100 (3.3 mm I.D. 3.6 m long) and a column molecular sieve-5A (3.3 mm I.D. 3.6 m long), connected with a thermal conductivity detector (accuracy of 1%) and hydrogen as the carrier gas, these conditions were previously described [14]. The amounts of carbon dioxide removed and oxygen produced was determined from samples taken from the gaseous phase of the system

(inlet and outlet) with a gas-tight syringe. The areas obtained using the integrator were compared with reference curves to determine the CO2 and O2 concentrations. 2.7.5. Analysis of total carbon Measurements of total carbon in the liquid and gaseous phases were carried out on a carbon analyzer TOC-VCSN (Shimatzu, Kyoto, Japan) with a normal sensibility catalyst (platinum on 1/800 Alumina Pellets) to measure total carbon (TC) and inorganic carbon (IC). Organic carbon (OC) was calculated by the difference between TC and IC. For the liquid phase measurements, 20 mL of the medium was ltered in a 0.2 mm lter and used for the analysis. In the analysis of total carbon, 27 mL samples were carried to the combustion tube at 680 8C, where the catalytic oxidization to CO2 occurred. For the analysis of inorganic carbon, samples of 27 mL reacted with hydrochloric acid 2 M to convert all the inorganic carbon into CO2. In both cases, the carbon dioxide was quantied by non-dispersive infrared absorption and the concentrations were calculated through analytical curves (peak area concentration) previously constructed with standard solutions of potassium hydrogen phthalate for TC and sodium hydrogen carbonate for IC. For gaseous measurements, gas samples were collected in the entrance ow and in the exit ow of the photobioreactor. For measurements of total carbon, 50 mL samples were injected and carried over a ow provided by an analytical O2 cylinder into the combustion tube (680 8C) and the samples were catalytic oxidized to CO2. Finally, to analyze inorganic carbon, 40 mL samples were injected and carried directly to the detector. They were quantied by non-dispersive infrared detector and the concentrations achieved with analytical curves of peak area concentration were made using the same conditions, with pure CO2. 2.7.6. Determination of elemental composition of the cells The composition of the elements of the Aphanothece microgeli cells was determined using a Perkin Elmer 2400 scopica Na CHNS/O element analyzer (accuracy of 0.4%). 2.7.7. Extraction and determination of total lipids, preparation of the fatty acid methyl esters, and determination of the fatty acid composition Lipids were extracted from the cells by the Bligh and Dyer method [13]. The method of Hartman and Lago [14] was used to saponify and esterify (methylation reaction) the dried lipid extract to obtain the fatty acid methyl esters (FAME) (biodiesel). Fatty acid composition was determined using a VARIAN 3600 CX gas chromatograph equipped with a ame ionization detector and a DB-FFAP model with a megabore column (30 m-long column with a diameter of 0.25 mm and a 0.25 mm thick lm). The carrier gas was nitrogen. The chromatographic conditions were previously described in Francisco et al. [15]. Most of the FAMEs were identied by comparison of the retention times with those of the standard (SupelcoTM 37 component FAME mix) and quantied by area normalization using Varian Star 4.51 software. The identity of unknown FAMEs was conrmed by GC-MS. Peaks were detected using a GCQ mass spectrometer (MS Saturn 2000 Varian) with electron impact ionization at 70 kV mass spectra of individual peaks were examined using Workstation 6.6 software. Identities were made based on the similarity of spectra between standard and candidate peaks. 2.7.8. Quality evaluation of biodiesel The quality properties of the biodiesel evaluated were ester content, cetane number, iodine value, degree of instauration and cold lter plugging point, determined according to the methodology proposed by Francisco et al. [15].

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3. Results and discussion 3.1. Potential renery gas streams Microalgae projects require large amounts of carbon dioxide, it is essential to nd suitable CO2 sources. The industrial application of photobioreactor technology depends on stationary sources of CO2 with temperatures and composition suitable to the growing to the target strains of microalgae. The purge gas, coming from the hydrogen purifying process, composed by 47% of CO2, 9% of CO, 20% of H2, 7% of N2 and 17% of CH4, emitted at 30 8C was the main source with potential for microalgae cultivation in the renery oil process considered (Fig. 2). Although it has several contaminants, it is emitted at a suitable temperature for biological processes. In addition, cyanobacteria are organisms known to have an adaptation capacity and after intensive acclimation step they were able to tolerate the industrial gaseous mixture. As it was not possible to install the photobioreactor in the oil renery, we simulated exactly the composition ratio of the industrial gas stream and diluted with air to obtain the following airstreams: purge gas at 5% of CO2 (5.0% of CO2, 0.9% of CO, 2.2% of H2, 1.8% of CH4, 18.7% of O2 and 70.7% of N2), purge gas at 15% of CO2 (15.0% of CO2, 2.9% of CO, 6.4% of H2, 5.4% of CH4, 14.3% of O2 and 55.1% of N2) and purge gas at 25% of CO2 (25.0% of CO2, 4.8% of CO, 10.6% of H2, 9.1% of CH4, 9.8% of O2 and 40.5% of N2). 3.2. Growth kinetics An effort to incorporate microalgae applications at wastewater treatment facilities is underway worldwide. Using the conditions previously established, growth kinetics was determined and results are shown in Table 2. In general, airstreams with 15% of carbon dioxide improved the growth kinetics [16], resulting in a maximum cellular concentration of 3.4 g/L, a maximum specic growth rate of 1.40 day1, a maximum pH value of 9.15, an average productivity of biomass of 0.47 g/L day and peak values of biomass productivity of 4.77 g/L day (HRT = 120 h). Comparatively, airstreams with 15% of carbon dioxide presented growth kinetics close to 50 and 35% superior to airstreams with 5 and 25% of carbon dioxide, respectively. The cultivation in the renery oil wastes at the best condition caused an increase in the cell growth rate, in comparison with cultivations with synthetic culture media and airstreams contaminated only with 15% of carbon dioxide (0.81 day1) [17]. This is a typical performance of the cyanobacterial cells that increase the metabolism to compensate for the adverse environment [7]. Comparatively, the biomass productivity was superior to the top three microalgae reported by Grifths and Harrison [18] in a compilation data of the 55 microalgae species. In this study, Amphora (average values for species AMPH027, AMPH045 and AMPH0546), Neochloris oleoabundans and Ankistrodesmus falcatus show biomass productivity values of 0.31, 0.37 and 0.45 g/L day, respectively. These values reect cultivations with pure CO2. On the other hand, Chiu et al. [9] reports maximum biomass productivity values of 0.37 g/L day for Chlorella sp. cultivated at 2% of ue gas from coke oven (25% of CO2, 4% of O2, 80 ppm of NO and 90 ppm of SO2). According these authors,

NATURAL GAS DESSULFURIZATION REACTOR SHIFT GAS COOLING WATER CONDENSED (rich in CO2) RECTIFICATION COLUMN HYDROGEN

PURIFICATION (PSA)
HYDROGEN PURIFIED PURGE GAS (CO2+CO+H2+N2+CH4+O2)

VAPOUR

WATER RETIFIED

WATER RETIFIED

GAS (CO2+ WATER VAPOUR)

20 ton/h T=30 C

5.5 ton/h T=140 C

Fig. 2. Fluxogram of the hydrogen generation unit at the renery oil plant.

some compounds such SOx and NOx inhibit the microalgal performance. However, after a lag period or a pre-adaptation, this limitation could be circumvented. 3.3. Carbon dioxide removal kinetics and oxygen production In order to test if the carbon metabolic performance of the geli was really due to cyanobacteria Aphanothece microscopica Na photosynthesis, a detailed study of carbon dioxide uptake and oxygen release from the photobioreactor was investigated. According to the Eriksen et al. [19] the theoretical photosynthetic quotient (ratio between the O2 release rate and the CO2 sequestration rate) is 0.73, where each 1 g of carbon dioxide consumed is correspondent to a release of 0.73 g of oxygen. The kinetic data for carbon dioxide removal expressed are showed in Table 3. Average CO2 elimination capacity of 9.82 mg/ L min with peaks of 22.91 mg/L min (HRT = 120 h) were obtained for purge gas at 15% of CO2, resulting in a global of CO2 removal efciency of 31.11%. Globally, 198 g of carbon dioxide were sequestered in this condition. This result is according to previous studies, where we determined that a 15% (v/v) content of CO2 in the air inlet improve carbon dioxide uptake performance by the geli in a bubble column cyanobacteria Aphanothece microscopica Na photobioreactor, considering contaminated airstreams only with CO2 [20]. Losses of the 70% of the carbon dioxide injected are typical of the conventional photobioreactor conguration [17,21], when high concentrations of CO2 are fed to the photobioreactor.

Table 2 geli cultivated with wastewater and purge gas from an oil renery. Growth kinetics for Aphanothece microscopica Na Parameter Xmax (g/L) mmax (day1) pHmax PX (average) (g/L day) PX (peak) (g/L day) Purge gas (5% CO2) 1.64 0.15 0.48 0.06 9.02 0.54 0.22 0.01 0.78 0.11 Purge gas (15% CO2) 3.40 0.57 1.40 0.02 9.15 0.45 0.47 0.04 4.77 0.28 Purge gas (25% CO2) 2.30 0.12 0.62 0.04 8.9 0.44 0.31 0.02 1.48 0.13

PX: biomass productivity, Xmax: maximum cellular concentration, pHmax: maximum pH value obtained in cultivation.

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E. Jacob-Lopes, T.T. Franco / Journal of CO2 Utilization xxx (2013) xxxxxx Table 3 Carbon dioxide removal kinetics and released oxygen. Parameter Average EC (mg/L min) Peak EC (mg/L min) Global RE (%) Global CO2 sequestered (g) Global released O2 (g) Purge gas (5% CO2) 2.75 0.13 6.44 0.38 26.51 2.12 55.64 2.78 41.17 1.23 Purge gas (15% CO2) 9.82 0.64 22.91 0.96 31.11 0.62 198.00 6.93 148.51 5.94 Purge gas (25% CO2) 7.94 0.48 18.54 1.05 14.90 0.71 160.27 8.49 119.40 4.17 5

EC: CO2 elimination capacity, RE: CO2 removal efciency.

Comparatively, the experiments conducted with 5% of CO2 were more efcient than that with 25% of CO2, if the removal efciency is considered as a principal performance indicator of the photobioreactor. However, removal efciency is not the best indicator of the photobioreactor performance because it varies with factors such as CO2 concentration and airow, and reects only the conditions of its measurement. On the other hand, the elimination capacity permits direct comparison of the results of the different photobioreactors systems, because the volume and ow are normalized and the CO2 input concentration is considered. The elimination capacity increased with the increasing concentration of carbon dioxide in the inlet when the system had a mass transfer limitation until it reached a maximum value, when it has a kinetic limitation. This maximum value is determined by the biodegradability of the compound [22]. In our system elimination capacity increased until 22.91 mg/L min with an inlet concentration of 15% of carbon dioxide, after that decrease (Fig. 3). Elimination capacity can only be equal or less than the mass loading rate. Under low load conditions, the elimination capacity essentially equals to the load, and the system is calculated to be at maximum removal efciency. By increasing the CO2 load on a system, a point will be reached where the mass loading overall mass loading rate will exceed the overall elimination capacity, reducing the CO2 removal efciencies. Efuent concentration is commonly used as the goal of regulatory compliance, when considering specic laws. At this moment, carbon dioxide does not have a target value. With basis on Kyoto protocol and the Clean Development Mechanism (CDM), elimination capacity should be considered as a main performance indicator of the photobioreactors because more carbon credits may be marketed. The global O2 released (Table 3) ranged from 41.17 to 148.51 g. These values are very close to be established by the theoretical

photosynthetic quotient. In this study, photosynthetic quotients close to 0.75 were obtained. 3.4. Carbon balance As the experimental photosynthetic quotient observed here was very close to 0.73 g, it is clear that Aphanotece microscopica geli is using carbon dioxide to construct chemicals and therefore, Na we used the carbon balance technique to investigate their location (liquid, solid and gas phases) and composition. A method was developed to nd, rstly, their location and later, their possible composition. Fig. 4 presents the representative carbon mass balance in the photobioreactor for a hydraulic retention time of 72 h, with purge gas diluted to 15% of CO2. As seen previously, the carbon losses represent 69% of the total CO2 amount injected in the system, indicating that 31% was biologically converted to chemicals. The carbon dioxide bioconverted in the system was distributed in organic carbon in the gaseous phase (92.0%), biomass in the solid phase (3.64%), inorganic carbon in the liquid phase (0.3%) and organic carbon in the liquid phase (0.05%). Carbon monoxide and methane were not considered in the balance. The gaseous phase was analyzed dynamically, and liquid and solid phases in an intermittent mode. For all conditions, the CO2 conversion into biomass represents a value of less than 5% of total sequestered. The CO2 conversion into bioproducts followed a bellshaped curve pattern for the conditions evaluated. These carbon dioxide conversion routes were previously described in our studies [12,17,23], where the biomass production represents a small fraction of the total carbon dioxide amount injected in the photobioreactor. The major fraction (92.0%) of the carbonaceous compounds formed is represented by the production and release of volatile organic compounds (VOCs). Microalgal cell can synthesize non-methane hydrocarbon, aldehydes and halocarbons [2427]. Initial characterization of the volatile fraction on photobioreactor (data not shown) by solid phase microextraction (SPME) indicate the predominance of the hydrocarbons up to 18 carbon atoms. Further investigation aimed to identify and characterize the individual VOC is necessary before the development of full-scale systems for the microalgal biorenery is achieved. If these volatile organic compounds formed are adequate and of high added value, this process could be considered an example of direct conversion of the CO2 into useful products, without the conventional intermediate step of biomass production. 3.5. Biodiesel production and biofuel quality The major obstacle for the commercialization of biodiesel is the high cost of oil feedstock. Instead of the vegetable oil industrially used, this process concentrated on biodiesel production from a microbial biomass, using oil-rening wastes. This is a new process in providing oil feedstock for biodiesel production, and is a future option for its application in the renery oil industry. Economically, however, is still a capital-intensive process. Table 4 shows the lipid content and lipid productivities of the geli. A lipid content of 17.1% was Aphanothece microscopica Na

Fig. 3. Elimination capacity (peak EC) vs. carbon dioxide load. Data points represent means (n = 4). Error bars indicate standard error from the mean.

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solid phase

biomass 1.95g

liquid phase

inorganic carbon 0.16g organic carbon 0.03g

liquid phase carbon from CO2 intermitent 171.79g PHOTOBIOREACTOR carbon from CO2 continuous 0.1029g

carbon loss 118.35g

carbon loss 0.1015g

gaseous phase

organic carbon 0.0013g

Fig. 4. Carbon mass balance in the photobioreactor (hydraulic retention time of 72 h).

Table 4 Lipid content and lipid productivities at purge gas with 15% of carbon dioxide. Parameter Airstream Purge gas (15% CO2) Lipid content (wt%) Lipid productivity (average) (g/L day) Lipid productivity (peak value) (g/L day) 17.1 1.02 0.08 0.04 0.81 0.28

Table 5 Fatty acid distribution in microalgal oil at purge gas with 15% of carbon dioxide. Fatty acids Saturated (SFA) Monounsaturated (MUFA) Polyunsaturated (PUFA) Percent (wt%) 49.13 2.01 35.82 1.18 14.77 0.31

measured in the biomass. Average lipid productivity and peak lipid productivity of 0.08 g/L day and 0.81 g/L day, respectively, were obtained. These values are higher than those obtained for geli cultivated on synthetic media Aphanothece microscopica Na and with airstreams contaminated only with carbon dioxide (15%, v/v) [15]. Lipid content for cultivations on renery wastes more than doubled in comparison with cultivations in synthetic sources of culture media and airstreams contaminated only with CO2 (8.0%). In addition, biomass productivity was slightly lower in such conditions, however, the equilibrium between lipid productivity and lipid content resulted in a lipid productivity 25% superior in cultivations on renery wastes. Environmental stresses, mainly nutritional stress are reported as the main causes to lipid content increase in microalgae cells. The handling of the nutritional conditions can to channel metabolic ux generated in the photobiosynthesis into lipid biosynthesis [28].

The fatty acid distribution of the oil extract is show in Table 5. geli cultivated on renery wastes has Aphanothece microscopica Na predominance in saturated (49.13%), monounsaturated (35.82%) and polyunsaturated fatty acids (14.77%), respectively. Of these fractions, thirty different fatty acids were identied, and heneicosanoic acid (C21:0), 38.65% and the elaidic acid (C18:1n9t), 28.9% were predominant. To assess the potential of biodiesel as a substitute of diesel fuel, the properties of biodiesel such as ester content, cetane number, iodine value, degree of insaturation and cold lter plugging point were determined. A comparison of properties of biodiesel from microalgal oil with US Standard [29], European Standard [30] and Brazilian Standard [31] is shown in Table 6. All parameters considered comply with the limits established by standards related to biodiesel quality. The physical and fuel properties of biodiesel, from microalgal oil in general, were comparable to those of

Table 6 Properties of biodiesel from microalgal oil (at purge gas with 15% of carbon dioxide), oilseeds and international standards. Properties EC (%) CN IV (gI2/100 g) DU (%) CFPP (8C) Microalgae 99.7 4.91 51.3 3.38 79.9 3.78 65.3 4.57 24.9 0.87 Palma 97.7 61 57 64.2 10 Soybeana 96.9 49 128 143.8 5 Rapeseeda 99.5 55 109 121.9 10 ASTM 6751 Min 47 EN 14214 Min 96.5 Min 51 Max 120 ANP 255 Min 45

EC: ester content, CN: cetane number, IV: iodine value, DU: degree of unsaturation, CFPP: cold lter plugging point. a Knothe [31], () not specied.

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vegetable sources. The biodiesel from microalgal oil showed a much higher value for cold lter plugging point of 24.9 8C. The European, American and Brazilian standards do not mention a lowtemperature parameter in their lists of specications, however, Knothe [32] suggests that each country specify certain temperature limits for different times of the year, depending on climate conditions. Biodiesel from microalgal oil have the highest CFPP because they are composed of long-carbon-chain saturated fatty acids. The longer the carbon chains in the biodiesel, the worse their low-temperature properties. Additives can be used to inhibit the agglomeration of crystals, thus lowering the point at which fuel lter plugging occurs [32]. 4. Conclusion The emerging microalgae industry continues its march toward industrial application. Wastewater associated to ue gases from rening oil industry are promising substrates for a microalgal biorenery platform. The liquid and gaseous wastes selected were suitable to support the photosynthetic cultivation of Aphanothece geli cyanobacterium, achieving substantial convermicroscopica Na sion rates of carbon dioxide into bioproducts (average elimination capacity of 9.82 mg/L min for purge gas at 15% of carbon dioxide). According to carbon balance analysis, volatile organic compounds (92.0%) are the main bioproducts formed and only a small fraction was converted into biomass (3.64%) in these conditions. Microalgal biodiesel production is a potential renewable energy source to be produced in microalgae integrated systems on oil reneries. In addition, volatile organic compounds can be valuable and protable bioproducts when efciently stored by suitable recovery operations. The integration of a microalgal biorenery to an oil renery plant seems to be feasible to improve the sustainability of the whole oil process. Acknowledgements leo Brasileiro S/ Funding for this research was provided by Petro A, Petrobras (Brazil). The authors are grateful to Dr. M.I. Queiroz (Federal University of Rio Grande, Brazil) for providing the cyanobacterium strain and to K. Modenesi and L. Furlan of Petrobras, for technical support.

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