Solar bioreactors used for the industrial production of microalgae.

Microalgae are excellent sources of biomass containing several important compounds for human and animal nutrition-proteins, lipids, polysaccharides, pigments and antioxidants as well as bioactive secondary metabolites. In addition, they have a great biotechnological potential for nutraceuticals, and pharmaceuticals as well as for CO2 sequestration, wastewater treatment, and potentially also biofuel and biopolymer production. In this review, the industrial production of the most frequently used microalgae genera-Arthrospira, Chlorella, Dunaliella, Haematococcus, Nannochloropsis, Phaeodactylum, Porphyridium and several other species is discussed as concerns the applicability of the most widely used large-scale systems, solar bioreactors (SBRs)-open ponds, raceways, cascades, sleeves, columns, flat panels, tubular systems and others. Microalgae culturing is a complex process in which bioreactor operating parameters and physiological variables closely interact. The requirements of the biological system-microalgae culture are crucial to select the suitable type of SBR. When designing a cultivation process, the phototrophic production of microalgae biomass, it is necessary to employ SBRs that are adequately designed, built and operated to satisfy the physiological requirements of the selected microalgae species, considering also local climate. Moreover, scaling up microalgae cultures for commercial production requires qualified staff working out a suitable cultivation regime. KEY POINTS: • Large-scale solar bioreactors designed for microalgae culturing. • Most frequently used microalgae genera for commercial production. • Scale-up requires suitable cultivation conditions and well-elaborated protocols.

Photosynthesis and biochemical characterization of the green alga Chlamydopodium fusiforme (Chlorophyta) grown in a thin-layer cascade.

Photosynthesis, growth and biochemical composition of the biomass of the freshwater microalga Chlamydopodium fusiforme cultures outdoors in a thin-layer cascade were investigated. Gross oxygen production measured off-line in samples taken from the outdoor cultures was correlated with the electron transport rate estimated from chlorophyll a fluorescence measurements. According to photosynthesis measurements, a mean of 38.9 ± 10.3 mol of photons were required to release one mole of O2, which is 4.86 times higher than the theoretical value (8 photons per 1 O2). In contrast, according to the fluorescence measurements, a mean of 11.7 ± 0.74 mol of photons were required to release 1 mol of O2. These findings indicate that fluorescence-based photosynthesis rates may not be fully replace oxygen measurements to evaluate the performance of an outdoor culture. Daily gross biomass productivity was 0.3 g DW L-1 day-1 consistently for 4 days. Biomass productivity was strongly affected by the suboptimal concentration at which the culture was operated and by the respiration rate, as the substantial volume of culture was kept in the dark (about 45% of the total volume). As the cells were exposed to excessive light, the photosynthetic activity was mainly directed to the synthesis of carbohydrates in the biomass. In the morning, carbohydrate content decreased because of the dark respiration. Per contra, protein content in the biomass was lower at the end of the day and higher in the morning due to carbohydrate consumption by respiration. The data gathered in these trials are important for the future exploitation of Chlamydopodium fusiforme as a potential novel species in the field of microalgae for the production of bio-based compounds.

Valuable pigments from microalgae: phycobiliproteins, primary carotenoids, and fucoxanthin.

Phycobiliproteins, carotenoids and fucoxanthin are photosynthetic pigments extracted from microalgae and cyanobacteria with great potential biotechnological applications, as healthy food colorants and cosmetics. Phycocyanin possesses a brilliant blue color, with fluorescent properties making it useful as a reagent for immunological essays. The most important source of phycocyanin is the cyanobacterium Arthrospira platensis, however, recently, the Rhodophyta Galdieria sulphuraria has also been identified as such. The main obstacle to the commercialization of phycocyanin is represented by its chemical instability, strongly reducing its shelf-life. Moreover, the high level of purity needed for pharmaceutical applications requires several steps which increase both the production time and cost. Microalgae (Chlorella, Dunaliella, Nannochloropsis, Scenedesmus) produce several light harvesting carotenoids, and are able to manage with oxidative stress, due to their free radical scavenging properties, which makes them suitable for use as source of natural antioxidants. Many studies focused on the selection of the most promising strains producing valuable carotenoids and on their extraction and purification. Among carotenoids produced by marine microalgae, fucoxanthin is the most abundant, representing more than 10% of total carotenoids. Despite the abundance and diversity of fucoxanthin producing microalgae only a few species have been studied for commercial production, the most relevant being Phaeodactylum tricornutum. Due to its antioxidant activity, fucoxanthin can bring various potential benefits to the prevention and treatment of lifestyle-related diseases. In this review, we update the main results achieved in the production, extraction, purification, and commercialization of these important pigments, motivating the cultivation of microalgae as a source of natural pigments.

Growth and photosynthetic performance of Nostoc linckia (formerly N. calcicola) cells grown in BG11 and BG110 media.

The biotechnological potential of Nostoc linckia as a biofertilizer and source of bioactive compounds makes it important to study its growth physiology and productivity. Since nitrogen is a fundamental component of N. linckia biomass, we compared the growth and biochemical composition of cultures grown in BG11 (i.e., in the presence of nitrate) and BG110 (in the absence of nitrate). Cultures grown in BG11 accumulated more cell biomass reaching a dry weight of 1.65 ± 0.06 g L-1, compared to 0.92 ± 0.01 g L-1 in BG110 after 240 h of culture. Biomass productivity was higher in culture grown in BG11 medium (average 317 ± 38 mg L-1 day-1) compared to that attained in BG110 (average 262 ± 37 mg L-1 day-1). The chlorophyll content of cells grown in BG11 increased continuously up to (39.0 ± 1.3 mg L-1), while in BG110 it increased much more slowly (13.6 ± 0.8 mg L-1). Biomass grown in BG11 had higher protein and phycobilin contents. However, despite the differences in biochemical composition and pigment concentration, between BG11 and BG110 cultures, both their net photosynthetic rates and maximum quantum yields of the photosystem II resulted in similar.

Photosynthesis Monitoring in Microalgae Cultures Grown on Municipal Wastewater as a Nutrient Source in Large-Scale Outdoor Bioreactors.

Microalgae cultures were used for a WW treatment to remediate nutrients while producing biomass and recycling water. In these trials, raceway ponds (RWPs; 1 and 0.5 ha) were located next to a municipal (WW) treatment plant in Mérida, Spain. The ponds were used for continuous, all-year-round microalgae production using WW as a source of nutrients. Neither CO2 nor air was supplied to cultures. The objective was to validate photosynthesis monitoring techniques in large-scale bioreactors. Various in-situ/ex-situ methods based on chlorophyll fluorescence and oxygen evolution measurements were used to follow culture performance. Photosynthesis variables gathered with these techniques were compared to the physiological behavior and growth of cultures. Good photosynthetic activity was indicated by the build-up of dissolved oxygen concentration up to 380% saturation, high photochemical yield (Fv/Fm = 0.62-0.71), and relative electron transport rate rETR between 200 and 450 µmol e- m-2 s-1 at midday, which resulted in biomass productivity of about 15-25 g DW m-2 day-1. The variables represent reliable markers reflecting the physiological status of microalgae cultures. Using waste nutrients, the biomass production cost can be significantly decreased for abundant biomass production in large-scale bioreactors, which can be exploited for agricultural purposes.

Effect of plate distance on light conversion efficiency of a Synechocystis culture grown outdoors in a multiplate photobioreactor.

In this work, the performance of a vertical multiplate photobioreactor is analyzed and presented. The photobioreactor consisted of 20 vertical plates (1 m2 each) connected by manifolds and a working volume of 1300 L. The total area occupied (footprint) was 10 m2, while the illuminated area was 40 m2, therefore the ratio of illuminated area to volume ratio was about 30 m-1. The performance of the photobioreactor was evaluated using a culture of Synechocystis PCC 6803, circulated by a centrifuge pump. The results showed that the amount of light captured by the photobioreactor at a plate spacing of 0.5 m was 90.2 % of the light incident on the horizontal surface, while at a plate spacing of 1.0 m, 50.3 % was captured. The corresponding biomass yield, calculated based on the ground area occupied by the reactor, was 26.0 g m-2 day-1 and 7.2 g m-2 day-1, when the plates were spaced at 0.5 m and 1.0 m respectively. Therefore, the light conversion efficiency calculated based on the ground area was significantly higher in the configuration with a plate spacing of 0.5 m, reaching 5.43 % based on PAR (photosynthetically active radiation), and 2.44 % based on solar radiation, giving a value 3.7 higher than when the plates were spaced 1.0 m apart. It was concluded that the light conversion efficiency might be further improved by reducing the plate spacing while also reducing the culture light path.

Influence of Different Light Sources on the Biochemical Composition of Arthrospira spp. Grown in Model Systems.

Arthrospira platensis and Arthrospira maxima are prokaryotic microalgae commercially marketed as spirulina. The pigments extracted from these algae are widely used for cosmetic and nutraceutical applications. This work aimed to evaluate the influence of three light-emitting lamps (white, orange and blue) on the growth and biomass composition of two strains of A. platensis (M2 and M2M) and one of A. maxima. The obtained data show strain- and light-dependent responses of the microalgae. In addition, white and orange lights led to a similar overall effect by increasing the levels of chlorophyll a and carotenoids. However, exposure to orange light resulted in the highest dry weight (5973.3 mg L-1 in M2M), whereas white light stimulated an increase in the carbohydrate fraction (up to 42.36 g 100 g-1 in A. maxima). Conversely, blue light led to a constant increase in the concentration of phycocyanin (14 g 100 g-1 in A. maxima) and a higher content of proteins in all strains. These results provide important environmental information for modulating the growth of different spirulina strains, which can be used to address the synthesis of biochemical compounds of strategic importance for the development of new nutraceutical foods.

In situ monitoring of chlorophyll a fluorescence in Nannochloropsis oceanica cultures to assess photochemical changes and the onset of lipid accumulation during nitrogen deprivation.

In situ chlorophyll a fluorescence measurements were applied to monitor changes in the photochemical variables of Nannochloropsis oceanica cultures under nitrogen-deplete and nitrogen-replete (control) conditions. In addition, growth, lipid, fatty acid, and pigment contents were also followed. In the control culture, growth was promoted along with pigment content, electron transport rate (ETR), and polyunsaturated fatty acids, while total lipid content and fatty acid saturation level diminished. Under nitrogen-deplete conditions, the culture showed a higher de-epoxidation state of the xanthophyll cycle pigments. Fast transients revealed a poor processing efficiency for electron transfer beyond QA , which was in line with the low ETR due to nitrogen depletion. Lipid content and the de-epoxidation state were the first biochemical variables triggered by the change in nutrient status, which coincided with a 20% drop in the in situ effective quantum yield of PSII (ΔF'/Fm '), and a raise in the Vj measurements. A good correlation was found between the changes in ΔF'/Fm ' and lipid content (r = -0.96, p < 0.01). The results confirm the reliability and applicability of in situ fluorescence measurements to monitor lipid induction in N. oceanica.

Isoprene Emission in Darkness by a Facultative Heterotrophic Green Alga.

Isoprene is a highly reactive biogenic volatile hydrocarbon that strongly influences atmospheric oxidation chemistry and secondary organic aerosol budget. Many phytoplanktons emit isoprene like terrestrial pants. Planktonic isoprene emission is stimulated by light and heat and is seemingly dependent on photosynthesis, as in higher plants. However, prominent isoprene-emitting phytoplanktons are known to survive also as mixotrophs and heterotrophs. Chlorella vulgaris strain G-120, a unicellular green alga capable of both photoautotrophic and heterotrophic growth, was examined for isoprene emission using GC-MS and real-time PTR-MS in light (+CO2) and in darkness (+glucose). Chlorella emitted isoprene at the same rate both as a photoautotroph under light, and as an exclusive heterotroph while feeding on exogenous glucose in complete darkness. By implication, isoprene synthesis in eukaryotic phytoplankton can be fully supported by glycolytic pathways in absence of photosynthesis, which is not the case in higher plants. Isoprene emission by chlorophyll-depleted mixotrophs and heterotrophs in darkness serves unknown functions and may contribute to anomalies in oceanic isoprene estimates.

Exopolysaccharide Features Influence Growth Success in Biocrust-forming Cyanobacteria, Moving From Liquid Culture to Sand Microcosms.

Land degradation in drylands is a drawback of the combined action of climate change and human activities. New techniques have been developed to induce artificial biocrusts formation as a tool for restoration of degraded drylands, and among them soils inoculation with cyanobacteria adapted to environmental stress. Improvement of soil properties by cyanobacteria inoculation is largely related to their ability to synthesize exopolysaccharides (EPS). However, cyanobacterial EPS features [amount, molecular weight (MW), composition] can change from one species to another or when grown in different conditions. We investigated the differences in growth and polysaccharidic matrix features among three common biocrust-forming cyanobacteria (Nostoc commune, Scytonema javanicum, and Phormidium ambiguum), when grown in liquid media and on sandy soil microcosms under optimal nutrient and water, in controlled laboratory conditions. We extracted and analyzed the released EPS (RPS) and sheath for the liquid cultures, and the more soluble or loosely-bound (LB) and the more condensed or tightly-bound (TB) soil EPS fractions for the sandy soil microcosms. In liquid culture, P. ambiguum showed the greatest growth and EPS release. In contrast, on the sandy soil, S. javanicum showed the highest growth and highest LB-EPS content. N. commune showed no relevant growth after its inoculation of the sandy soil. A difference was observed in terms of MW distribution, showing that the higher MW of the polymers produced by P. ambiguum and S. javanicum compared to the polymers produced by N. commune, could have had a positive effect on growth for the first two organisms when inoculated on the sandy soil. We also observed how both RPS and sheath fractions reflected in the composition of the soil TB-EPS fraction, indicating the role in soil stabilization of both the released and the cell attached EPS. Our results indicate that the features of the polysaccharidic matrix produced by different cyanobacteria can influence their growth success in soil. These results are of great relevance when selecting suitable candidates for large-scale cyanobacteria applications in soil restoration.

Acclimation strategy of Rhodopseudomonas palustris to high light irradiance.

The ability of Rhodopseudomonas palustris cells to rapidly acclimate to high light irradiance is an essential issue when cells are grown under sunlight. The aim of this study was to investigate the photo-acclimation process in Rhodopseudomonas palustris 42OL under different culturing conditions: (i) anaerobic (AnG), (ii) aerobic (AG), and (iii) under H2-producing (HP) conditions both at low (LL) and high light (HL) irradiances. The results obtained clearly showed that the photosynthetic unit was significantly affected by the light irradiance at which Rp. palustris 42OL was grown. The synthesis of carotenoids was affected by both illumination and culturing conditions. At LL, lycopene was the main carotenoid synthetized under all conditions tested, while at HL under HP conditions, it resulted the predominant carotenoid. Oppositely, under AnG and AG at HL, rhodovibrin was the major carotenoid detected. The increase in light intensity produced a deeper variation in light-harvesting complexes (LHC) ratio. These findings are important for understanding the ecological distribution of PNSB in natural environments, mostly characterized by high light intensities, and for its growth outdoors.

H2 production in Rhodopseudomonas palustris as a way to cope with high light intensities.

The ability of coping with the damaging effects of high light intensity represents an essential issue when purple non-sulfur bacteria (PNSB) are grown under direct sunlight for photobiological hydrogen production. This study was aimed at investigating whether H2 photo-evolution could represent, for Rhodopseudomonas palustris 42OL, a safety valve to dissipate an excess of reducing power generated under high light intensities. The physiological status of this strain was assessed under anaerobic (AnG) and aerobic (AG) growing conditions and under H2-producing (HP) conditions at low and high light intensities. The results obtained clearly showed that Fv/Fm ratio was significantly affected by the light intensity under which R. palustris 42OL cells were grown, under either AnG or AG conditions, while, under HP, it constantly remained at its highest value. The increase in light intensity significantly increased the H2 production rate, which showed a positive correlation with the maximum electron transfer rate (rETRmax). These findings are important for optimization of hydrogen production by PNSB under solar light.

Effect of high pH on growth of Synechocystis sp. PCC 6803 cultures and their contamination by golden algae (Poterioochromonas sp.).

Culturing cyanobacteria in a highly alkaline environment is a possible strategy for controlling contamination by other organisms. Synechocystis PCC 6803 cells were grown in continuous cultures to assess their growth performance at different pH values. Light conversion efficiency linearly decreased with the increase in pH and ranged between 12.5 % (PAR) at pH 7.5 (optimal) and decreased to 8.9 % at pH 11.0. Photosynthetic activity, assessed by measuring both chlorophyll fluorescence and photosynthesis rate, was not much affected going from pH 7.5 to 11.0, while productivity, growth yield, and biomass yield on light energy declined by 32, 28, and 26 % respectively at pH 11.0. Biochemical composition of the biomass did not change much within pH 7 and 10, while when grown at pH 11.0, carbohydrate content increased by 33 % while lipid content decreased by about the same amount. Protein content remained almost constant (average 65.8 % of dry weight). Cultures maintained at pH above 11.0 could grow free of contaminants (protozoa and other competing microalgae belonging to the species of Poterioochromonas).

A bioenergetic assessment of photosynthetic growth of Synechocystis sp. PCC 6803 in continuous cultures.

BACKGROUND: Synechocystis sp. PCC 6803, a model organism used for bioenergy and bioplastic production, was grown in continuous culture to assess its most important bioenergetic parameters. RESULTS: Biomass yield on light energy of 1.237 g mol photons(-1) and maintenance energy requirement of 0.00312 mol photons g(-1) h(-1) were calculated. This corresponded to a light conversion efficiency of 12.5 %, based on the model of Pirt which was about 35 % lower than the theoretical one based on the stoichiometric equation for the formation of biomass on carbon dioxide, water, and nitrate. The maximum F v/F m ratio recorded in the Synechocystis cultures was 0.57; it progressively declined to 0.45 as the dilution rate increased. An over-reduction of reaction centers at a high dilution rate was also recorded, together with an increased VJ phase for the chlorophyll fluorescence transient. In contrast, the chlorophyll optical cross section increased by about 40 % at the fastest dilution rate, and compensated for the decline in F v/F m, thus resulting in a constant total photosynthesis rate (photosynthesis plus respiration). Chlorophyll content was maximum at the lowest dilution rate and was 48 % lower at the highest one, while phycocyanin, and total carotenoids decreased by about 42 % and 37 %, respectively. Carotenoid analysis revealed increased echinenone, zeaxanthin, and myxoxanthophyll contents as the dilution rate increased (40.6, 63.8 and 35.5 %, respectively, at the fastest dilution rate). A biochemical analysis of the biomass harvested at each different dilution rates showed no changes in the lipid content (averaging 11.2 ± 0.6 % of the dry weight), while the protein content decreased as the dilution rate increased, ranging between 60.7 ± 1.1 and 72.6 ± 0.6 %. Amino acids pattern did not vary with the dilution rate. Carbohydrate content ranged from 9.4 to 16.2 % with a mean value of 11.2 ± 1.4 %. CONCLUSIONS: The present work provides useful information on the threshold of light conversion efficiency in Synechocystis, as well as basic bioenergetic parameters that will be helpful for future studies related to its genetic transformation and metabolic network reconstruction.

Advances in the biotechnology of hydrogen production with the microalga Chlamydomonas reinhardtii.

Biological hydrogen production is being evaluated for use as a fuel, since it is a promising substitute for carbonaceous fuels owing to its high conversion efficiency and high specific energy content. The basic advantages of biological hydrogen production over other "green" energy sources are that it does not compete for agricultural land use, and it does not pollute, as water is the only by-product of the combustion. These characteristics make hydrogen a suitable fuel for the future. Among several biotechnological approaches, photobiological hydrogen production carried out by green microalgae has been intensively investigated in recent years. A select group of photosynthetic organisms has evolved the ability to harness light energy to drive hydrogen gas production from water. Of these, the microalga Chlamydomonas reinhardtii is considered one of the most promising eukaryotic H2 producers. In this model microorganism, light energy, H2O and H2 are linked by two excellent catalysts, the photosystem 2 (PSII) and the [FeFe]-hydrogenase, in a pathway usually referred to as direct biophotolysis. This review summarizes the main advances made over the past decade as an outcome of the discovery of the sulfur-deprivation process. Both the scientific and technical barriers that need to be overcome before H2 photoproduction can be scaled up to an industrial level are examined. Actual and theoretical limits of the efficiency of the process are also discussed. Particular emphasis is placed on algal biohydrogen production outdoors, and guidelines for an optimal photobioreactor design are suggested.

Scenedesmus incrassatulus CLHE-Si01: a potential source of renewable lipid for high quality biodiesel production.

The potential of microalgal oil from Scenedesmus incrassatulus as a feedstock for biodiesel production was studied. Cell concentration of S. incrassatulus and lipid content obtained during mixotrophic growth were 1.8 g/L and 19.5 ± 1.5% dry cell weight, respectively. The major components of biodiesel obtained from S. incrassatulus oil were methyl palmitate (26%) and methyl linoleate (49%), which provided a strong indication of high quality biodiesel. Fuel properties were determined by empirical equations and found to be within the limits of biodiesel standard ASTM D6751 and EN 14214. The quality properties of the biodiesel were high cetane number (62), low density (803 kg/m(3)), low viscosity (3.78 mm(2)/s), oxidation stability (9h) and cold filter plugging point (-4°C). Hence, S. incrassatulus has potential as a feedstock for the production of excellent quality biodiesel.

Unusual catalysts from molasses: synthesis, properties and application in obtaining biofuels from algae.

Acid catalysts were prepared by sulfonation of carbon materials obtained from the pyrolysis of sugar beet molasses, a cheap, viscous byproduct in the processing of sugar beets into sugar. Conditions for the pyrolysis of molasses (temperature and time) influenced catalyst performance; the best combination came from pyrolysis at low temperature (420 °C) for a relatively long time (8-15 h), which ensured better stability of the final material. The most effective molasses catalyst was highly active in the esterification of fatty acids with methanol (100 % yield after 3 h) and more active than common solid acidic catalysts in the transesterification of vegetable oils with 25-75 wt % of acid content (55-96 % yield after 8 h). A tandem process using a solid acid molasses catalyst and potassium hydroxide in methanol was developed to de-acidificate and transesterificate algal oils from Chlamydomonas reinhardtii, Nannochloropsis gaditana, and Phaeodactylum tricornutum, which contain high amounts of free fatty acids. The amount of catalyst required for the de-acidification step was influenced by the chemical composition of the algal oil, thus operational conditions were determined not only in relation to free fatty acids content in the oil, but according to the composition of the lipid extract of each algal species.

Outdoor H2 production in a 50-L tubular photobioreactor by means of a sulfur-deprived culture of the microalga Chlamydomonas reinhardtii.

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.

Sustained H2 production in a Chlamydomonas reinhardtii D1 protein mutant.

In the present investigation, a detailed biochemical analysis of the high H2 producer D1 protein mutant strain L159I-N230Y of Chlamydomonas reinhardtii, carrying a double amino acid substitution, was made. The leucine residue L159 was replaced by isoleucine, and the N230 asparagine was replaced by tyrosine. The performance of this strain was compared to that of the cc124 strain. The mutant showed a sustained capacity to donate electrons by means of direct biophotolysis for H2 production, as demonstrated by the higher efficiency of utilization of the hydrogenase enzyme when carried out under anaerobic conditions. The latter property was maintained also under sulfur deprivation. Furthermore, when compared to the cc124, the mutant showed a higher amount of D1 protein content, a higher carbohydrate storage capacity and a sustained PSII direct contribution to the H2 production during sulfur deprivation. The addition of DCMU to the cells showed that as much as 7.0 mL H2 liter of culture h⁻¹ were produced by means of direct biophotolysis. The maximum apparent light-to-hydrogen conversion efficiency expressed on PAR (photosynthetically active radiation) reached 3.22%, while PSII efficiency to perform direct biophotolysis was calculated to be 2.03%. These values are significantly higher than what has been reported in the literature.

Preliminary investigation on the production of fuels and bio-char from Chlamydomonas reinhardtii biomass residue after bio-hydrogen production.

The aim of this work was to investigate the potential conversion of Chlamydomonas reinhardtii biomass harvested after hydrogen production. The spent algal biomass was converted into nitrogen-rich bio-char, biodiesel and pyrolysis oil (bio-oil). The yield of lipids (algal oil), obtained by solvent extraction, was 15 ± 2% w/w(dry-biomass). This oil was converted into biodiesel with a 8.7 ± 1% w/w(dry-biomass) yield. The extraction residue was pyrolysed in a fixed bed reactor at 350 °C obtaining bio-char as the principal fraction (44 ± 1% w/w(dry-biomass)) and 28 ± 2% w/w(dry-biomass) of bio-oil. Pyrolysis fractions were characterized by elemental analysis, while the chemical composition of bio-oil was fully characterized by GC-MS, using various derivatization techniques. Energy outputs resulting from this approach were distributed in hydrogen (40%), biodiesel (12%) and pyrolysis fractions (48%), whereas bio-char was the largest fraction in terms of mass.