Microalgae–bacteria symbiosis in microalgal growth and biofuel production: a review

Photosynthetic microalgae can capture solar energy and convert it to bioenergy and biochemical products. In nature or industrial processes, microalgae live together with bacterial communities and may maintain symbiotic relationships. In general interactions, microalgae exude dissolved organic carbon that becomes available to bacteria. In return, the bacteria remineralize sulphur, nitrogen and phosphorous to support the further growth of microalgae. In specific interactions, heterotrophic bacteria supply B vitamins as organic cofactors or produce siderophores to bind iron, which could be utilized by microalgae, while the algae supply fixed carbon to the bacteria in return. In this review, we focus on mutualistic relationship between microalgae and bacteria, summarizing recent studies on the mechanisms involved in microalgae–bacteria symbiosis. Symbiotic bacteria on promoting microalgal growth are described and the relevance of microalgae–bacteria interactions for biofuel production processes is discussed. Symbiotic microalgae–bacteria consortia could be utilized to improve microalgal biomass production and to enrich the biomass with valuable chemical and energy compounds. The suitable control of such biological interactions between microalgae and bacteria will help to improve the microalgae-based biomass and biofuel production in the future.     

Life-cycle assessment of microalgae culture coupled to biogas production

Due to resource depletion and climate change, lipid-based algal biofuel has been pointed out as an interesting alternative because of the high productivity of algae per hectare and per year and its ability to recycle CO₂ from flue gas. Another option for taking advantage of the energy content of the microalgae is to directly carry out anaerobic digestion of raw algae in order to produce methane and recycle nutrients (N, P and K). In this study, a life-cycle assessment (LCA) of biogas production from the microalgae Chlorella vulgaris is performed and the results are compared to algal biodiesel and to first generation biodiesels. These results suggest that the impacts generated by the production of methane from microalgae are strongly correlated with the electric consumption. Progresses can be achieved by decreasing the mixing costs and circulation between different production steps, or by improving the efficiency of the anaerobic process under controlled conditions. This new bioenergy generating process strongly competes with others biofuel productions.     

Mutual influence of light and CO2 on carbon sequestration via cultivating mixotrophic alga Auxenochlorella protothecoides UMN280 in an organic carbon-rich wastewater

This study focusses on the assimilation of carbon in concentrated municipal wastewater rich in organic carbon using the mixotrophic microalga Auxenochlorella protothecoides UMN280 with the addition of supplemental CO₂. The entire growth period of A. protothecoides UMN280 can be characterized by three phases: first, a phase where algae grew in a mixotrophic-dominated mode; second, a transition phase; and last, a phase where algae grew in a photoautotrophic-dominated mode. In this study, it was found that light intensity had a strong effect on algal biomass production; the culture system would transfer from a mixotrophic-dominated mode to a photoautotrophic-dominated mode quicker under higher light intensities. The addition of CO₂ exhibited an important role in the photoautotrophic-dominated cultivation stage. At certain level of irradiance and certain range of CO₂ injection rate, higher CO₂ injection rate would result in a higher level of carbon fixation. It is clearly beneficial to inject exogenous CO₂ in the mixotrophic wastewater algae production system when a light source is available, such as during daylight hours.     

Photochemical and Microbial Degradation of 2,4,5-Trichloroaniline in a Freshwater Lake

Surface waters from a eutrophic lake in northern Georgia were incubated with ¹⁴C-labeled 2,4,5-trichloroaniline to study the disappearance of the parent compound and production of ¹⁴CO₂. There was no degradation of the compound in the dark. Under 12 h of sunlight and 12 h of darkness, 28% of the trichloroaniline was degraded in both poisoned and untreated samples. Mineralization after 24 h in poisoned and untreated lake water was 5.5 and 6.8%, respectively. Thus, 81% of the mineralization was attributable to photochemical processes, and 19% was attributable to microbial processes. Most biological mineralization was due to microbes of bacterial size (<1.0 μm). Approximately 90% of the trichloroaniline bioaccumulated was associated with organisms larger than 1.0 μm, e.g., algae. When algae were removed by filtration, the amount of trichloroaniline mineralized increased to 9.4%, compared with 6.8% in the presence of algae. The excretion of organic compounds by algae may have inhibited bacterial mineralization of photoproducts.     

Potential of diatom consortium developed by nutrient enrichment for biodiesel production and simultaneous nutrient removal from waste water

Because of the decreasing fossil fuel supply and increasing greenhouse gas (GHG) emissions, microalgae have been identified as a viable and sustainable feedstock for biofuel production. The major effect of the release of wastewater rich in organic compounds has led to the eutrophication of freshwater ecosystems. A combined approach of freshwater diatom cultivation with urban sewage water treatment is a promising solution for nutrient removal and biofuel production. In this study, urban wastewater from eutrophic Hussain Sagar Lake was used to cultivate a diatom algae consortium, and the effects of silica and trace metal enrichment on growth, nutrient removal, and lipid production were evaluated. The nano-silica-based micronutrient mixture Nualgi containing Si, Fe, and metal ions was used to optimize diatom growth. Respectively, N and P reductions of 95.1% and 88.9%, COD and BOD reductions of 91% and 51% with a biomass yield of 122.5 mg L^?1 day^?1 and lipid productivity of 37 mg L^?1 day^?1 were observed for cultures grown in waste water using Nualgi. Fatty acid profiles revealed 13 different fatty acids with slight differences in their percentage of dry cell weight (DCW) depending on enrichment level. These results demonstrate the potential of diatom algae grown in wastewater to produce feedstock for renewable biodiesel production. Enhanced carbon and excess nutrient utilization makes diatoms ideal candidates for co-processes such as CO₂ sequestration, biodiesel production, and wastewater phycoremediation.     

CYANOPHYTA-BACTERIA SYSTEMS: EFFECTS OF ADDED CARBON COMPOUNDS OR PHOSPHATE ON ALGAL GROWTH AT LOW NUTRIENT CONCENTRATIONS1

Planktonic blue-green algae are known to be always associated with bacteria. Earlier work has shown that the addition of a bacteria-assimilable carbon source to a normal Zehnder-Gorham culture medium (No. 11) will produce enhanced growth of these algae when atmospheric CO₂ has become the limiting factor. In new work, phosphate-rich culture media were diluted so that they simulated nutrient levels found in the Great Lakes, e.g., Lake Erie. At these low concentrations and when atmospheric CO₂ was not available in a sufficient supply, the addition of sucrose to either a 1/100 or a 1/1000 diluted Z-G, medium (10 mg or 2 mg of sucrose, respectively) also produced enhanced, growth of the tested blue-green algae. The stimulation of algal growth was apparently due to an increased bacterial production of CO₂ and possibly other carbon compounds approaching the composition of the CO₂ molecule. The literature suggests that during vigorous algal growth in lakes, atmospheric CO₂ will be severely limiting. Productive lakes always contain nonliving organic matter. The presence of bacteria-assimilable matter is probably one of the important factors leading to algal bloom.     

Algal photosynthetic aeration increases the capacity of bacteria to degrade organics in wastewater

Wastewater treatment is an energy-intensive process and a net emitter of greenhouse gas emissions. A large fraction of these emissions is due to intensive aeration of aerobic bacteria to facilitate break-down of organic compounds. Algae can generate dissolved oxygen at levels in excess of saturation, and therefore hold the potential to partially displace or complement mechanical aeration in wastewater treatment processes. The objective of this study was to develop an internally consistent experimental and modeling approach to test the hypothesis that algal photosynthetic aeration can speed the removal of organic constituents by bacteria. This framework was developed using a simplified wastewater treatment process consisting of a model bacteria (Escherichia coli), a model algae (Auxenochlorella protothecoides), and a single carbon source that was consumable by bacteria only. This system was then tested both with and without the presence of algae. A MATLAB model that considered mass transfer and biological kinetics was used to estimate the production and consumption of O₂ and CO₂ by algae and bacteria. The results indicated that the presence of algae led to 18–66% faster removal of COD by bacteria, and that roughly one-third of biochemical oxygen demand was offset by algal photosynthetic aeration.     

Atmospheric impact of bioenergy based on perennial crop (reed canary grass,Phalaris arundinaceae,L.) cultivation on a drained boreal organic soil

Marginal organic soils, abundant in the boreal region, are being increasingly used for bioenergy crop cultivation. Using long‐term field experimental data on greenhouse gas (GHG) balance from a perennial bioenergy crop [reed canary grass (RCG), Phalaris arundinaceae L.] cultivated on a drained organic soil as an example, we show here for the first time that, with a proper cultivation and land‐use practice, environmentally sound bioenergy production is possible on these problematic soil types. We performed a life cycle assessment (LCA) for RCG on this organic soil. We found that, on an average, this system produces 40% less CO₂‐equivalents per MWh of energy in comparison with a conventional energy source such as coal. Climatic conditions regulating the RCG carbon exchange processes have a high impact on the benefits from this bioenergy production system. Under appropriate hydrological conditions, this system can even be carbon‐negative. An LCA sensitivity analysis revealed that net ecosystem CO₂ exchange and crop yield are the major LCA components, while non‐CO₂ GHG emissions and costs associated with crop production are the minor ones. Net bioenergy GHG emissions resulting from restricted net CO₂ uptake and low crop yields, due to climatic and moisture stress during dry years, were comparable with coal emissions. However, net bioenergy emissions during wet years with high net uptake and crop yield were only a third of the coal emissions. As long‐term experimental data on GHG balance of bioenergy production are scarce, scientific data stemming from field experiments are needed in shaping renewable energy source policies.     

Metagenome Survey of a Multispecies and Alga-Associated Biofilm Revealed Key Elements of Bacterial-Algal Interactions in Photobioreactors

Photobioreactors (PBRs) are very attractive for sunlight-driven production of biofuels and capturing of anthropogenic CO₂. One major problem associated with PBRs however, is that the bacteria usually associated with microalgae in nonaxenic cultures can lead to biofouling and thereby affect algal productivity. Here, we report on a phylogenetic, metagenome, and functional analysis of a mixed-species bacterial biofilm associated with the microalgae Chlorella vulgaris and Scenedesmus obliquus in a PBR. The biofilm diversity and population dynamics were examined through 16S rRNA phylogeny. Overall, the diversity was rather limited, with approximately 30 bacterial species associated with the algae. The majority of the observed microorganisms were affiliated with Alphaproteobacteria, Betaproteobacteria, and Bacteroidetes. A combined approach of sequencing via GS FLX Titanium from Roche and HiSeq 2000 from Illumina resulted in the overall production of 350 Mbp of sequenced DNA, 165 Mbp of which was assembled in larger contigs with a maximum size of 0.2 Mbp. A KEGG pathway analysis suggested high metabolic diversity with respect to the use of polymers and aromatic and nonaromatic compounds. Genes associated with the biosynthesis of essential B vitamins were highly redundant and functional. Moreover, a relatively high number of predicted and functional lipase and esterase genes indicated that the alga-associated bacteria are possibly a major sink for lipids and fatty acids produced by the microalgae. This is the first metagenome study of microalga- and PBR-associated biofilm bacteria, and it gives new clues for improved biofuel production in PBRs.     

Algal biofuels

The world is facing energy crisis and environmental issues due to the depletion of fossil fuels and increasing CO₂ concentration in the atmosphere. Growing microalgae can contribute to practical solutions for these global problems because they can harvest solar energy and capture CO₂ by converting it into biofuel using photosynthesis. Microalgae are robust organisms capable of rapid growth under a variety of conditions including in open ponds or closed photobioreactors. Their reduced biomass compounds can be used as the feedstock for mass production of a variety of biofuels. As another advantage, their ability to accumulate or secrete biofuels can be controlled by changing their growth conditions or metabolic engineering. This review is aimed to highlight different forms of biofuels produced by microalgae and the approaches taken to improve their biofuel productivity. The costs for industrial-scale production of algal biofuels in open ponds or closed photobioreactors are analyzed. Different strategies for photoproduction of hydrogen by the hydrogenase enzyme of green algae are discussed. Algae are also good sources of biodiesel since some species can make large quantities of lipids as their biomass. The lipid contents for some of the best oil-producing strains of algae in optimized growth conditions are reviewed. The potential of microalgae for producing petroleum related chemicals or ready-make fuels such as bioethanol, triterpenic hydrocarbons, isobutyraldehyde, isobutanol, and isoprene from their biomass are also presented.     

Converting carbon dioxide to high value-added products: Microalgae-based green biomanufacturing

Today's world faces the dual pressure of carbon dioxide (CO₂) emission reduction and an energy crisis. Microalgae, which can use solar energy to convert CO₂ to organic matter, have emerged as a promising and renewable cell factory for producing nutrients, biofuels, and various high value-added compounds (HVACs). They possess numerous advantages, such as high photosynthetic efficiency, fast growth rate, and use of agro-industrial waste and nonagricultural land for cultivation. Microalgae can also effectively remove eutrophic elements (e.g., nitrogen and phosphorus) from wastewater and atmospheric pollutants (e.g., SO_x and NO_x) from flue gas, thus providing great environmental benefits. However, microalgae-based production often faces low productivity, limiting applicability in industrial settings. Genetic and metabolic modifications of certain microalgal strains have proven effective in improving productivity. Here, we review the latest developments regarding the microalgae-based production of platform compounds, biofuels, and other HVACs. Although still in the early exploration stage, the rapid development of gene editing tools, a deeper understanding of the metabolic pathways of microalgae and their regulatory mechanisms, and further optimization of cultivation procedures and photosynthetic efficiency can eventually enable the launch of microalgae-based biomanufacturing for green industrial production. Therefore, this technology is strategically important for solving the current energy crisis problems of excessive CO₂ emissions and environmental pollution. This review provides information about the advancement and development of microalgae-based production over the past two decades and discusses possible future directions in the field.     

微藻在CO2生物捕集及废水生态修复领域的研究进展

温室效应、水资源短缺和能源危机是21世纪人类面临的三大挑战。微藻是一种水生植物,在CO₂减排、废水生态修复及生物能源领域已成为全球研究热点。综述了微藻在CO₂生物捕集和废水生态修复的应用研究进展。微藻生物柴油现已成为全球研发热点,但研究主要集中在某个单元的最优化设计,而对各单元之间相互作用和耦合的重要性缺乏充分认知,提出了将CO₂生物捕集、废水生态修复、生物柴油制备、藻渣替代水煤浆与煤共气化的理念,这对微藻生物过程的高效全局优化和环境综合治理具有重要意义,是未来我国发展低碳经济的有效途径,并在此基础上对微藻产业规模化的未来核心研究方向进行了展望。     

微藻-细菌共生体系在废水处理中的应用

在微藻-细菌协同共生的过程中,藻类光合作用释放的氧气被异养微生物利用来矿化水体中的污染物,细菌呼吸为藻类提供二氧化碳作为碳源。近年来,藻类-细菌协同共生体系在污水处理中的应用得到了广泛的研究。本文重点综述了菌藻协同共生体系中微藻与细菌之间的三种相互作用,以及菌藻协同共生体系在废水处理中的应用。菌藻协同共生体系中的微藻与细菌通过营养交换、信号转导及基因转移等相互作用实现共赢。该体系广泛用于处理富营养化、重金属、药物、多环芳烃(polycyclicaromatic hydrocarbons,PAHs)、石油烃化合物等难降解的有机污染的水体。对于氮、磷等营养物质的去除,其主要机理涉及同化作用、厌氧氨氧化作用、硝化与反硝化作用、磷酸化作用等。对重金属、药物、石油烃化合物及其他有机化合物的去除机制主要是生物吸附、生物富集及细胞内外的生物降解。     

Microalgae: a robust “green bio-bridge” between energy and environment

Microalgae are a potential candidate for biofuel production and environmental treatment because of their specific characteristics (e.g. fast growth, carbon neutral, and rich lipid accumulations). However, several primary bottlenecks still exist in current technologies, including low biomass conversion efficiency, bio-invasion from the external environment, limited or costly nutrient sources, and high energy and capital input for harvest, and stalling its industrial progression. Coupling biofuel production with environmental treatment renders microalgae a more feasible feedstock. This review focuses on microalgae biotechnologies for both bioenergy generation and environmental treatment (e.g. CO₂ sequestration and wastewater reclamation). Different intelligent technologies have been developed, especially during the last decade, to eliminate the bottlenecks, including mixotrophic/heterotrophic cultivation, immobilization, and co-cultivation. It has been realized that any single purpose for the cultivation of microalgae is not an economically feasible option. Combinations of applications in biorefineries are gradually reckoned to be necessary as it provides more economically feasible and environmentally sustainable operations. This presents microalgae as a special niche occupier linking the fields of energy and environmental sciences and technologies. The integrated application of microalgae is also proven by most of the life-cycle analysis studies. This study summarizes the latest development of primary microalgal biotechnologies in the two areas that will bring researchers a comprehensive view towards industrialization with an economic perspective.     

Depth-Dependent Mineral Soil CO 2 Production Processes: Sensitivity to Harvesting-Induced Changes in Soil Climate

Forest harvesting induces a step change in the climatic variables (temperature and moisture), that control carbon dioxide (CO ₂) production arising from soil organic matter decomposition within soils. Efforts to examine these vertically complex relationships in situ within soil profiles are lacking. In this study we examined how the climatic controls on CO ₂ production change within vertically distinct layers of the soil profile in intact and clearcut forest soils of a humid temperate forest system of Atlantic Canada. We measured mineral soil temperature (0, 5, 10, 20, 50 and 100 cm depth) and moisture (0–15 cm and 30–60 cm depth), along with CO ₂ surface efflux and subsurface concentrations (0, 2.5, 5, 10, 20, 35, 50, 75 and 100 cm depth) in 1 m deep soil pits at 4 sites represented by two forest-clearcut pairs over a complete annual cycle. We examined relationships between surface efflux at each site, and soil heat, moisture, and mineral soil CO ₂ production. Following clearcut harvesting we observed increases in temperature through depth (1–2°C annually; often in excess of 4°C in summer and spring), alongside increases in soil moisture (30%). We observed a systematic breakdown in the expected exponential relationship between CO ₂ production and heat with mineral soil depth, consistent with an increase in the role moisture plays in constraining CO ₂ production. These findings should be considered in efforts to model and characterize mineral soil organic matter decomposition in harvested forest soils.     

Partitioning of CH4 and CO2 Production Originating from Rice Straw,Soil and Root Organic Carbon in Rice Microcosms

Flooded rice fields are an important source of the greenhouse gas CH₄. Possible carbon sources for CH₄ and CO₂ production in rice fields are soil organic matter (SOM), root organic carbon (ROC) and rice straw (RS), but partitioning of the flux between the different carbon sources is difficult. We conducted greenhouse experiments using soil microcosms planted with rice. The soil was amended with and without ¹³C-labeled RS, using two ¹³C-labeled RS treatments with equal RS (5 g kg⁻¹ soil) but different δ¹³C of RS. This procedure allowed to determine the carbon flux from each of the three sources (SOM, ROC, RS) by determining the δ¹³C of CH₄ and CO₂ in the different incubations and from the δ¹³C of RS. Partitioning of carbon flux indicated that the contribution of ROC to CH₄ production was 41% at tillering stage, increased with rice growth and was about 60% from the booting stage onwards. The contribution of ROC to CO₂ was 43% at tillering stage, increased to around 70% at booting stage and stayed relatively constant afterwards. The contribution of RS was determined to be in a range of 12–24% for CH₄ production and 11–31% for CO₂ production; while the contribution of SOM was calculated to be 23–35% for CH₄ production and 13–26% for CO₂ production. The results indicate that ROC was the major source of CH₄ though RS application greatly enhanced production and emission of CH₄ in rice field soil. Our results also suggest that data of CH₄ dissolved in rice field could be used as a proxy for the produced CH₄ after tillering stage.     

Carbon dioxide utilisation of Dunaliella tertiolecta for carbon bio-mitigation in a semicontinuous photobioreactor

Bio-fixation of carbon dioxide (CO₂) by microalgae has been recognised as an attractive approach to offset anthropogenic emissions. Biological carbon mitigation is the process whereby autotrophic organisms, such as microalgae, convert CO₂ into organic carbon and O₂ through photosynthesis; this process through respiration produces biomass. In this study Dunaliella tertiolecta was cultivated in a semicontinuous culture to investigate the carbon mitigation rate of the system. The algae were produced in 1.2-L Roux bottles with a working volume of 1 L while semicontinuous production commenced on day 4 of cultivation when the carbon mitigation rate was found to be at a maximum for D. tertiolecta. The reduction in CO₂ between input and output gases was monitored to predict carbon fixation rates while biomass production and microalgal carbon content are used to calculate the actual carbon mitigation potential of D. tertiolecta. A renewal rate of 45 % of flask volume was utilised to maintain the culture in exponential growth with an average daily productivity of 0.07 g L^?1 day^?1. The results showed that 0.74 g L^?1 of biomass could be achieved after 7 days of semicontinuous production while a total carbon mitigation of 0.37 g L^?1 was achieved. This represented an increase of 0.18 g L^?1 in carbon mitigation rate compared to batch production of D. tertiolecta over the same cultivation period.     

Phycoremediation coupled production of algal biomass,harvesting and anaerobic digestion: Possibilities and challenges

Biogas produced from anaerobic digestion is a versatile and environment friendly fuel which traditionally utilizes cattle dung as the substrate. In the recent years, owing to its high content of biodegradable compounds, algal biomass has emerged as a potential feedstock for biogas production. Moreover, the ability of algae to treat wastewater and fix CO₂ from waste gas streams makes it an environmental friendly and economically feasible feedstock. The present review focuses on the possibility of utilizing wastewater as the nutrient and waste gases as the CO₂ source for algal biomass production and subsequent biogas generation. Studies describing the various harvesting methods of algal biomass as well as its anaerobic digestion have been compiled and discussed. Studies targeting the most recent advancements on biogas enrichment by algae have been discussed. Apart from highlighting the various advantages of utilizing algal biomass for biogas production, limitations of the process such as cell wall resistivity towards digestion and inhibitions caused due to ammonia toxicity and the possible strategies for overcoming the same have been reviewed. The studies compiled in the present review indicate that if the challenges posed in translating the lab scale studies on phycoremediation and biogas production to pilot scale are overcome, algal biogas could become the sustainable and economically feasible source of renewable energy.     

Energy Demands of Nitrogen Supply in Mass Cultivation of Two Commercially Important Microalgal Species, <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> and <Emphasis Type="Italic">Dunaliella tertiolecta</Emphasis>

Mass culture of microalgae is a potential alternative to cultivation of terrestrial crops for bioenergy production. However, microalgae require nitrogen fertiliser in quantities much higher than plants, and this has important consequences for the energy balance of these systems. The effect of nitrogen fertiliser supplied to microalgal bubble-column photobioreactor cultures was investigated using different nitrogen sources (nitrate, urea, ammonium) and culture conditions (air, 12% CO₂). In 20 L cultivations, maximum biomass productivity for Chlorella vulgaris cultivated using nitrate and urea was 0.046 and 0.053 g L⁻¹ day⁻¹, respectively. Maximum biomass productivity for Dunaliella tertiolecta cultivated using nitrate, urea and ammonium was 0.033, 0.038 and 0.038 g L⁻¹ day⁻¹, respectively. In intensive bubble-column photobioreactors using 12% CO₂, maximum productivity reached 0.60 and 0.83 g L⁻¹ day⁻¹ for C. vulgaris and D. tertiolecta, respectively. Recycling of nitrogen within the photobioreactor system via algal exudation of nitrogenous compounds and bacterial activity was identified as a potentially important process. The energetic penalty incurred by supply of artificial nitrogen fertilisers, phosphorus, power and CO₂ to microalgal photobioreactors was investigated, although analysis of all energy burdens from biomass production to usable energy carriers was not conducted. After subtraction of the power, nitrogen and phosphorus energy burdens, maximum net energy ratios for C. vulgaris and D. tertiolecta cultivated in bubble columns were 1.82 and 2.10. Assuming CO₂ was also required from a manufactured source, the net energy ratio decreased to 0.09 and 0.11 for C. vulgaris and D. tertiolecta, so that biomass production in this scenario was unsustainable. Although supply of nitrogen is unlikely to be the most energetically costly factor in sparged photobioreactor designs, it is still a very significant penalty. There is a need to optimise both cultivation strategies and recycling of nitrogen in order to improve performance. Data are supported by measurements including biochemical properties (lipid, protein, heating value) and bacterial number by epifluorescence microscopy.     

Optimization of CO2 concentration and light intensity for biodiesel production by Chlorella vulgaris FACHB-1072 under nitrogen deficiency with phosphorus luxury uptake

Microalgal biodiesel is an alternative bioenergy for the future. Nitrogen deprivation is usually used to increase lipid content in microalgae, however, it also lowers biomass production, resulting in not much increase of lipid productivity. Our previous study found that phosphorus played an important role in enhancing biodiesel productivity of C. vulgaris FACHB-1072 under nitrogen deficient condition. The aim of this study was to optimize two significant parameters of CO₂ concentration (0.03, 4, 6, 12 %) and light intensity (40, 120, 200 μmol photons m^-2 s^-1) with respect to biodiesel productivity and P uptake rate of C. vulgaris FACHB-1072. It was found that the optimized conditions were 4 % CO₂ concentration and 200 μmol photons m^-2 s^-1 light intensity. The maximum biodiesel productivity was 34.56 mg L^-1 day^-1; 2.7 times higher than the control (nutrient sufficient condition). Phosphorus was accumulated as polyphosphate and its maximum uptake rate was 2.08 mg L^-1 day^-1; twice that of the control. After optimization, the performances under nitrogen deficiency were significantly better compared with those under nitrogen sufficiency, which were rarely reported in literature. Our findings suggest a great potential to combine phosphorus removal from wastewater with biodiesel production via microalgae.