Prospects of biodiesel production from microalgae in India

Energy is essential and vital for development, and the global economy literally runs on energy. The use of fossil fuels as energy is now widely accepted as unsustainable due to depleting resources and also due to the accumulation of greenhouse gases in the environment. Renewable and carbon neutral biodiesel are necessary for environmental and economic sustainability. Biodiesel demand is constantly increasing as the reservoir of fossil fuel are depleting. Unfortunately biodiesel produced from oil crop, waste cooking oil and animal fats are not able to replace fossil fuel. The viability of the first generation biofuels production is however questionable because of the conflict with food supply. Production of biodiesel using microalgae biomass appears to be a viable alternative. The oil productivity of many microalgae exceeds the best producing oil crops. Microalgae are photosynthetic microorganisms which convert sunlight, water and CO₂ to sugars, from which macromolecules, such as lipids and triacylglycerols (TAGs) can be obtained. These TAGs are the promising and sustainable feedstock for biodiesel production. Microalgal biorefinery approach can be used to reduce the cost of making microalgal biodiesel. Microalgal-based carbon sequestration technologies cover the cost of carbon capture and sequestration. The present paper is an attempt to review the potential of microalgal biodiesel in comparison to the agricultural crops and its prospects in India.     

Advances and perspectives in using microalgae to produce biodiesel

Carbon-neutral renewable liquid biofuels are needed to displace petroleum-derived transport fuels in the near future – which contribute to global warming and are of a limited availability. A promising alternative is conveyed by microalgae, the oil content of which may exceed 80% (w/w_DW) – as compared with 5% of the best agricultural oil crops. However, current implementation of microalga-based systems has been economically constrained by their still poor volumetric efficiencies – which lead to excessively high costs, as compared with petrofuel prices. Technological improvements of such processes are thus critical – and this will require a multiple approach, both on the biocatalyst and bioreactor levels. Several bottlenecks indeed exist at present that preclude the full industrial exploitation of microalgal cells: the number of species that have been subjected to successful genetic transformation is scarce, which hampers a global understanding (and thus a rational design) of novel blue-biotechnological processes; the mechanisms that control regulation of gene expression are not fully elucidated, as required before effective bioprocesses based on microalgae can be scaled-up; and new molecular biology tools are needed to standardize genetic modifications in microalgae – including efficient nuclear transformation, availability of promoter or selectable marker genes, and stable expression of transgenes. On the other hand, a number of pending technological issues are also present: the relatively low microalga intrinsic lipid productivity; the maximum cell concentration attainable; the efficiency of harvest and sequential recovery of bulk lipids; and the possibility of by-product upgrade. This review briefly covers the state of the art regarding microalgae toward production of biofuels, both from the point of view of the microalgal cell itself and of the supporting bioreactor; and discusses, in a critical manner, current limitations and promising perspectives in this field.     

Potential of microalgae oil from Dunaliella tertiolecta as a feedstock for biodiesel

Alternative, non-food based biomass fuel feedstock development is vital for our national security, economy and the environment. Microalgae are among the most promising of these alternatives. Microalgal cell growth rates and metabolic products are affected by a combination of environmental parameters. In this work, the influences of light source, light intensity, CO₂ concentration, and photoperiod on the growth of Dunaliella tertiolecta (D. tertiolecta) were studied. The effects of these environmental parameters on the lipid content and fatty acid composition of D. tertiolecta were also investigated. Red light-emitting diodes (LEDs), white LEDs, and fluorescent lights were all found to be effective for algal growth. Increasing light intensity resulted in significantly more rapid algal growth, and increasing the period of light also significantly increased biomass productivity. Similar growth rates were observed for 2%, 4%, and 6% CO₂-concentrations. The different light sources and intensities were found to have no significant effect on FAME composition of D. tertiolecta. Methyl linolenate and methyl palmitate were found to be the major components of FAME produced from D. tertiolecta oil. D. tertiolecta and its derived oils should be a suitable feedstock for biofuel production.     

Biomass and lipid productivities of marine microalgae isolated from the Persian Gulf and the Qeshm Island

In this work, the screening of 147 microalgal strains from the Persian Gulf and the Qeshm Island (Iran) were done in order to choose the best ones, in terms of growth (biomass) rate and lipid content for biodiesel production. A methodology, combining experiments in lab-scale and pilot plant (open pond) used to produce and evaluate biomass and lipid productivity is presented for the systematic investigation of the potential of different microalgae species. The culture conditions, including photo flux (180 ??E m⁻² s⁻¹), photoperiod (12 h light/dark), temperature (25 °C), pH (≈8), air (carbon dioxide) and growth medium, were kept constant for all experiments. Microalgae were screened in two stages using optical density (for evaluation of biomass concentration) and Nile red and gas chromatography (for determination of lipid content and fatty acid fractions). In general, maximum specific growth rate and the maximum biomass productivity were obtained after 8-12-day culture. Nannochloropsis sp. and Neochloris sp. were selected from the marine microalgal culture collection, due to their high biomass (50 and 21.7 g L⁻¹, respectively) and oil content (52% and 46%, respectively). If the purpose is to produce biodiesel only from one species, Nannochloropsis sp. presented the most adequate fatty acid profile, namely linolenic and other polyunsaturated fatty acids. However, the microalgae Chlorella sp. can also be used if associated with other microalgal oils. In addition, selected strains could be potent candidates for commercial production in the open pond culture.     

Technoeconomic analysis of an integrated microalgae photobioreactor, biodiesel and biogas production facility

As fossil fuel prices increase and environmental concerns gain prominence, the development of alternative fuels from biomass has become more important. Biodiesel produced from microalgae is becoming an attractive alternative to share the role of petroleum. Currently it appears that the production of microalgal biodiesel is not economically viable in current environment because it costs more than conventional fuels. Therefore, a new concept is introduced in this article as an option to reduce the total production cost of microalgal biodiesel. The integration of biodiesel production system with methane production via anaerobic digestion is proved in improving the economics and sustainability of overall biodiesel stages. Anaerobic digestion of microalgae produces methane and further be converted to generate electricity. The generated electricity can surrogate the consumption of energy that require in microalgal cultivation, dewatering, extraction and transesterification process. From theoretical calculations, the electricity generated from methane is able to power all of the biodiesel production stages and will substantially reduce the cost of biodiesel production (33% reduction). The carbon emissions of biodiesel production systems are also reduced by approximately 75% when utilizing biogas electricity compared to when the electricity is otherwise purchased from the Victorian grid. The overall findings from this study indicate that the approach of digesting microalgal waste to produce biogas will make the production of biodiesel from algae more viable by reducing the overall cost of production per unit of biodiesel and hence enable biodiesel to be more competitive with existing fuels.     

The effects of ultraviolet radiation on growth,biomass, lipid accumulation and biodiesel properties of microalgae

The effect of UV light on growth, biomass, lipid accumulation and biodiesel properties of microalgae was studied. A Microalgae strain Chlorella sorokiniana UUIND6 was cultivated for 14 days as under LED light (Control) and microalgae were exposed to UV light (280–320 nm) in the middle of the photoperiod for 3 days. The growth rate of microalgae was analyzed by spectrophotometer and cell counting, while oil accumulation was analyzed by improved Nile red method. Results showed that microalgae under UV light treated algal cells showed less growth. FAMEs profile of UV treated algal cells mainly contains hexadecanoic acid (C16), stearic acid (C18) fatty acids. PUFA found in very less amount in UV treated cells as compared to control.     

Neochloris oleoabundans grown on anaerobically digested dairy manure for concomitant nutrient removal and biodiesel feedstock production

Microalgae have been investigated as a promising biodiesel feedstock; however, large-scale production is not currently cost-competitive with petroleum diesel, and its environmental impacts have received little attention. Using wastewater to supply nutrients for algal growth obviates synthetic fertilizer use, provides on-site nutrient removal, and reduces greenhouse gas emissions. In this work, anaerobically digested dairy manure was used to grow the oleaginous green alga Neochloris oleoabundans. In batch culture experiments with both synthetic media and anaerobic digester effluent, N. oleoabundans assimilated 90-95% of the initial nitrate and ammonium after 6 d and yielded 10-30% fatty acid methyl esters on a dry weight basis. Cellular lipid content and the N concentration in the growth media were inversely correlated. In addition, the proportion of polyunsaturated fatty acids (i.e. C16:3, C18:2, and C18:3) decreased with N concentration over time while the proportion of C18:1 fatty acid increased. Although N deficiency is likely the primary driver behind lipid accumulation, the influence of culture pH confounded results and requires further study. Other living microorganisms in the digester effluent were not observed to affect algal growth and lipid productivity, though the breakdown of organic nitrogen may have hindered lipid accumulation traditionally achieved through the manipulation of synthetic media. This work highlights the potential for waste-grown mono-algal cultures to produce high quality biodiesel while accomplishing simultaneous wastewater treatment.     

Biodiesel from microalgae – Life cycle assessment and recommendations for potential improvements

Microalgae are considered as one of the potential major source of biofuel for the future. However, their environmental benefit is still unclear and many scientific publications provide contradictory results. Here we perform the Life Cycle Assessment of the production and combustion of 1 MJ of algal methylester. The system under consideration uses standard open raceways under greenhouses. Lipid extraction and transesterification are carried out on a humid paste produced by centrifugation. Our environmental and energetic analysis shows that improving the energy balance is clearly the key priority to make microalgal cultivation sustainable and to reduce its greenhouse gas (GHG) emissions. To achieve significant reduction of the GHG emissions, most of the studies of the literature focus on technological breakthroughs, especially at the production step. However, since a large fraction of environmental impacts and especially GHG emissions do not occur directly at the production facility but stem from the production of the electricity required for producing, harvesting and transforming algae, it seems relevant to question the source of electricity as well as algae production technology. We consider a scenario where up to 45% of electricity was produced by a local renewable source and then we compare it to the improvements resulting from technological breakthroughs resulting in higher microalgal productivity or biomass concentration. It turns out that increasing the yield only drastically reduces the climate change for low starting productivity. The climate change is always significantly reduced by the use of local renewable electricity. It is therefore wiser to increase biomass productivity to easily achievable values (10–15 gm⁻² d⁻¹), and then radically change improvements pathways by considering the composition of the electricity mix used for example. At least, it must be underlined that the introduction of renewable electricity also affect energetic efficiency, leading to a positive cumulative energy balance due to better energetic ratios.     

Search for new strains of microalgae-producers of lipids from natural sources for biodiesel production

Biomass of high-yielding strains of phototrophic microorganisms actively accumulating lipids is a promising non-traditional raw material for bioenergy including the production of biodiesel. In this study, we present results of searching for new strains of microalgae-producers of lipids from hot springs. Within the framework of research, the primary screening of water for the presence of lipid - accumulative microalgae was carried out with the help of qualitative reaction with lipid-specific dyes, as well as 5 axenic isolates of microalgae with stable growth were identified in the laboratory and their productivity and fatty acid composition were studied. The isolated strains were identified as Chlorella vulgaris sp-1, Ankistrodesmus sp-21, Scеnеdеsmus obliquus sp-21, Chlorella pyrenoidosa sp-13 and Chlamydomonas sp-22. The obtained data showed that the isolated strains determined by biomass in the range 1.3 g/l to 1.81 g/l. As a result of the research, it was established that the highest content of lipids was observed in the strains Chlorella vulgaris sp-1 and Scеnеdеsmus obliquus sp-21, which is 28.7 and 29.8% of the cell dry weight, respectively. The analysis of the fatty acid composition of the cells showed that the largest mass fraction of saturated and monounsaturated fatty acids was found in strain Scеnеdеsmus obliquus sp-21 - 61.9%. In the result, Scеnеdеsmus obliquus sp-21 strain isolated from thermal sources was selected as a promising candidate for biodiesel production.     

Oxidative stress-tolerant microalgae strains are highly efficient for biofuel feedstock production on wastewater

Nutrient-rich wastewater may provide a sustainable means to cultivate microalgal biomass for biofuel use, yet many microalgal strains are very sensitive to wastewater due to toxicity caused by abiotic and biotic stresses. Naturally adapted strains that can efficiently grow in wastewater effluent are therefore of interest, however, the mechanisms by which such strains tolerate wastewater conditions are unknown. This study isolated indigenous chlorophyte microalgae strains from a municipal secondary wastewater effluent tank. The strains were identified by molecular phylogenetics and characterised by their ability to utilise exogenous organic carbon sources for mixotrophic growth and on the basis of oxidative stress tolerance, in order to elucidate the mechanisms of wastewater adaptation. Two of the strains, identified as Chlorella luteoviridis and Parachlorella hussii, could grow very well in raw wastewater due to their substantial tolerance to oxidative stress, which is highly induced by the wastewater environment. These strains exhibited high ascorbate peroxidase activity allowing increased scavenging of reactive oxygen species compared to strains that are not well adapted to the wastewater conditions. Both strains displayed high biomass and lipid productivity values in wastewater effluent. The accumulated lipids were suitable for biodiesel usage with characteristics equivalent to palm oil- and sunflower oil-derived biodiesel. The strains were also efficient in nutrient remediation from the wastewater. These results demonstrate the potential of these two strains for future biofuel applications coupled to wastewater remediation and highlight the importance of oxidative stress tolerance as a key indicator of efficient wastewater growth.     

Role of hydrogen in improving performance and emission characteristics of homogeneous charge compression ignition engine fueled with graphite oxide nanoparticle-added microalgae biodiesel/diesel blends

The development of low-temperature combustion models combined with the use of biofuels has been considered as an efficient strategy to reduce pollutant emissions like CO, HC. NO_x, and smoke. Indeed, Homogeneous Charge Compression Ignition (HCCI) is the new approach to drastically minimize NO_x emissions and smoke owing to the lower cylinder temperature and a higher rate of homogeneous A/F mixture as compared to compression ignition (CI) engines. The present research deal with the behavior analysis of a CI engine powered by diesel, Euglena Sanguinea (ES), and their blends (ES20D80, ES40D60, ES60D40, ES80D20). The experimental results revealed the highest brake thermal efficiency for ES20D80 although it decreased by 4.1% compared to diesel at normal mode. The average drop in HC, CO, and smoke was 2.1, 2.3, and 5.7% for ES20D80 as opposed to diesel fuel. Therefore, in the next stage, ES20D80 with various concentrations of graphite oxide (GO) nanoparticle (20, 40, 60, and 80 ppm) was chosen to carry out experiments in the HCCI mode, in which hydrogen gas was induced along with air through the intake pipe at a fixed flow rate of 3 lpm for the enrichment of the air-fuel mixture. As a result, the combination of hydrogen-enriched gas and GO-added ES20D80 in the HCCI mode showed similar performance to the CI engine but registered a major reduction of NO_x and smoke emissions, corresponding to 75.24% and 53.07% respectively, as compared to diesel fuel at normal mode.     

Effective extraction of microalgae lipids from wet biomass for biodiesel production

Producing biodiesel from lipid extracted from microalgae is a promising approach for sustainable fuel production. However, this approach is not yet commercialized due to the high costs of upstream processes that are associated with the time consuming and/or energy intensive drying, and lipid extraction processes. In this study, the possibility of avoiding the drying process, and extracting the lipid directly from the wet concentrated cells, using enzymatic disruption to enhance the extraction, has been tested. Results showed that lysozyme and cellulase were both efficient in disrupting cell walls and enhancing lipid extraction from wet samples, with highest lipid extraction yield of 16.6% achieved using lysozyme. The applicability of using supercritical CO₂ (SC-CO₂) in extracting lipid from wet biomass was also tested and the highest yield of 12.5% was achieved using lysozyme. In addition, a two-step culturing process was applied, using Scenedesmus sp., to combine both high biomass growth and lipid content. The strain was able to increase its biomass productivity in the first stage, reaching 174 mg l⁻¹ d⁻¹, with almost constant lipid content. In the second stage, the lipid content was enhanced by six-fold after three weeks of nitrogen starvation, but with lower biomass productivity.     

Anaerobic digestion of microalgae residues resulting from the biodiesel production process

The recovery of methane from post transesterified microalgae residues has the potential to improve the renewability of the ‘microalgae biomass to biodiesel’ conversion process as well as reduce its cost and environmental impact. This paper deals with the anaerobic digestion of microalgae biomass residues (post transesterification) using semi-continuously fed reactors. The influence of substrate loading concentrations and hydraulic retention times on the specific methane yield of the anaerobically digested microalgae residues was investigated. The co-digestion of the microalgae residues with glycerol as well as the influence of temperature was also examined. It was found that the hydraulic retention period was the most significant variable affecting methane production from the residues, with periods (>5 days) corresponding to higher energy recovery. The methane yield was also improved by a reduction in the substrate loading rates, with an optimum substrate carbon to nitrogen ratio of 12.44 seen to be required for the digestion process.     

Characterization of microalgal species isolated from fresh water bodies as a potential source for biodiesel production

Due to increasing oil prices and climate change concerns, biodiesel has gained attention as an alternative energy source. Biodiesel derived from microalgae is a potentially renewable and carbon–neutral alternative to petroleum fuels. One of the most important decisions in obtaining oil from microalgae is the choice of algal species to use. Eight microalgae from a total of 33 isolated cultures were selected based on their morphology and ease of cultivation. Five cultures were isolated from river and identified as strains of Scenedesmus obliquus YSR01, Nitzschia cf. pusilla YSR02, Chlorella ellipsoidea YSR03, S. obliquus YSR04, and S. obliquus YSR05, and three were isolated from wastewater and identified as S. obliquus YSW06, Micractinium pusillum YSW07, and Ourococcus multisporus YSW08, based on LSU rDNA (D1-D2) and ITS sequence analyses. S. obliquus YSR01 reached a growth rate of 1.68 ± 0.28 day⁻¹ at 680_nm and a biomass concentration of 1.57 ± 0.67 g dwt L⁻¹, with a high lipid content of 58 ± 1.5%. Under similar environmental conditions, M. pusillum reached a growth rate of 2.3 ± 0.55 day⁻¹ and a biomass concentration of 2.28 ± 0.16 g dwt L⁻¹, with a relatively low lipid content of 24 ± 0.5% w/w. The fatty acid compositions of the studied species were mainly myristic, palmitic, palmitoleic, oleic, linoleic, g-linolenic, and linolenic acids. Our results suggest that S. obliquus YSR01 can be a possible candidate species for producing oils for biodiesel, based on its high lipid and oleic acid contents.     

Microalgae for biodiesel production and other applications: A review

Sustainable production of renewable energy is being hotly debated globally since it is increasingly understood that first generation biofuels, primarily produced from food crops and mostly oil seeds are limited in their ability to achieve targets for biofuel production, climate change mitigation and economic growth. These concerns have increased the interest in developing second generation biofuels produced from non-food feedstocks such as microalgae, which potentially offer greatest opportunities in the longer term. This paper reviews the current status of microalgae use for biodiesel production, including their cultivation, harvesting, and processing. The microalgae species most used for biodiesel production are presented and their main advantages described in comparison with other available biodiesel feedstocks. The various aspects associated with the design of microalgae production units are described, giving an overview of the current state of development of algae cultivation systems (photo-bioreactors and open ponds). Other potential applications and products from microalgae are also presented such as for biological sequestration of CO₂, wastewater treatment, in human health, as food additive, and for aquaculture.     

Evaluation and optimization of two stage sequential in situ transesterification process for fatty acid methyl ester quantification from microalgae

This study demonstrates a direct transesterification (DT) method for reliable quantification of microalgal lipid. Primary screening of various transesterification methods and the types of biomass (wet, oven dried and lyophilized) were performed with heterotrophically grown Chlorella sp. FC2 IITG which revealed two stage DT with lyophilized biomass using NaOH in first stage and H₂SO₄ in second stage as the best combination with fatty acid methyl ester (FAME) yield of 39.17% (w/w, dry cell weight). Further optimization of transesterification parameters for selected method using response surface methodology, predicted the optimum values for catalyst to biomass ratio 0.67 (w/w) and 2.07 (v/w), methanol to biomass ratio 49.51 (v/w) and 61.07 (v/w) and reaction time 19.33 (min) and 10 (min) for first and second stages respectively. The optimum conditions showed 462.6% and 445.4% increment in FAME yield when compared with Bligh and Dyer method for Chlorella sp. FC2 IITG and Chlorella sorokiniana FC6 IITG respectively with highest transesterification efficiency of 98.96%. Improved transesterification efficiency of two stage DT was attributed to efficient destabilization of cell wall as confirmed by scanning electron microscopic imaging. FAME produced via DT of Chlorella sp. FC2 IITG satisfied most of the biodiesel properties as per ASTM D6751 and hence, could be an alternative to petro-diesel.     

Effect of hydroxy and hydrogen gas addition on diesel engine fuelled with microalgae biodiesel

Owing to high growth rate, being non-edible, and environmental friendliness; microalgae is a promising third generation biodiesel raw material. In this study, hydrogen and hydroxy gas aspirated compression ignition engine which was fuelled with microalgae biodiesel and low sulphur diesel fuel blend were investigated in order to evaluate their combined effect. The results showed that the brake power and torque output of the test engine decreased with microalgae biodiesel usage. Moreover, microalgae biodiesel addition results in lower carbon monoxide and nitrogen oxides emissions, and higher carbon dioxide. The introduction of hydrogen and hydroxy gas compensated the decrement of torque and power output and increment of carbon dioxide emission. The study enlightened that usage of microalgae biodiesel with hydrogen and hydroxy gas addition is a very promising combination from the environmental viewpoint.     

Effects of flocculants on lipid extraction and fatty acid composition of the microalgae Nannochloropsis oculata and Thalassiosira weissflogii

The aim of this study was to investigate the possible interference of anionic and cationic flocculants in the lipid extraction and fatty acid profiles of two species of marine microalgae: Nannochloropsis oculata and Thalassiosira weissflogii. Cells were grown in batch cultures (f/2 medium, salinity of 28, temperature of 20 °C, light intensity of 40 ??mol photons  m^-2 s^-1 and 12/12 h L/D photoperiod) and concentrated using sodium hydroxide (control), sodium hydroxide and the anionic polyacrylamide flocculant Magnafloc^® LT-25 (APF treatment) and sodium hydroxide plus the cationic polyacrylamide flocculant Flopam^® (CPF treatment). There were no statistically significant differences among treatments with respect to lipid extraction for both species. However, N. oculata which presented higher percentages of C16:0, C16:1 and C20:5 fatty acids showed an increase of C14:0 and a decrease of C20:5 with the use of anionic flocculant. Additionally, T. weissflogii which had high percentages of C16:0, C16:1, C16:3 and C20:5, showed a decrease of C18:0 and C18:1n9c when both flocculants were used and a small decrease of C16:0 in the APF treatment. The results indicate that the choice of flocculant should be based on the level of saturation desirable, i.e., if the goal is to produce more stable biodiesel, with low percentage unsaturated fatty acids, then anionic flocculants should be used. On the other hand, if the aim is to produce unsaturated fatty acids for commercial uses in the pharmacy or food industries, then anionic polymers should be avoided.     

Characterization and identification of lipid-producing microalgae species isolated from a freshwater lake

Microalgal lipids are the oils of the future for sustainable biodiesel production. One of the most important decisions in obtaining oil from microalgae is the choice of species. A total of 45 algal cultures were isolated from a freshwater lake at Wonju, South Korea. Five microalgal isolates were selected based on their morphology and ease of cultivation under our test conditions. These cultures were identified as strains of Scenedesmus obliquus YSL02, Chlamydomonas pitschmannii YSL03, Chlorella vulgaris YSL04, S. obliquus YSL05, and Chlamydomonas mexicana YSL07 based on microscopic examination and LSU rDNA (D1-D2) sequence analysis. S. obliquus YSL02 reached a higher biomass concentration (1.84 ± 0.30 g L⁻¹) with a lower lipid content (29% w/w), than did Chla. pitschmannii YSL03 (maximum biomass concentration of 1.04 ± 0.09 with a 51% lipid content). Our results suggest that Chla. pitschmannii YSL03 is appropriate for producing biodiesel based on its high lipid content and oleic acid proportion.