Comparative profiling of Chlorella vulgaris cells, extracts, and intact chloroplasts using electron transfer matrix-assisted laser desorption/ionization mass spectrometry (ET-MALDI-MS).

The intricate composition of microalgal pigments plays a crucial role in various biological processes, from photosynthesis to biomarker identification. Traditional pigment analysis methods involve complex extraction techniques, posing challenges in maintaining analyte integrity. In this study, we employ Electron Transfer Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (ET-MALDI-MS) to compare the pigmentary profiles of Chlorella vulgaris intact cells, chloroplasts, and solvent extracts. We aim to obtain comprehensive extracts rich in polar and non-polar compounds using ultrasound-assisted and supercritical fluid extraction methods. Additionally, intact chloroplasts are isolated using a lysis buffer and sucrose density gradient centrifugation. Our ET-MALDI-MS analysis reveals distinct compositional differences, highlighting the impact of extraction protocols on microalgal pigment identification. We observe prominent signals corresponding to radical cations of key pigments, including chlorophylls and carotenoids, which are crucial for C. vulgaris identification. Furthermore, ET-MALDI-MS facilitates the identification of specific lipids within chloroplast membranes and other organelles. This study underscores the rapid and precise nature of ET-MALDI-MS in microalgal biomarker analysis, providing valuable insights into phytoplankton dynamics, trophic levels, and environmental processes. C. vulgaris emerges as a promising model for studying pigment composition and membrane lipid diversity, enhancing our understanding of microalgal ecosystems.

Chlorella vulgaris-mediated bioremediation of food and beverage wastewater from industries in Mexico: Results and perspectives towards sustainability and circular economy.

The food and beverage industries in Mexico generate substantial effluents, including nejayote, cheese-whey, and tequila vinasses, which pose significant environmental challenges due to their extreme physicochemical characteristics and excessive organic load. This study aimed to assess the potential of Chlorella vulgaris in bioremediating these complex wastewaters while also producing added-value compounds. A UV mutagenesis treatment (40 min) enhanced C. vulgaris adaptability to grow in the effluent conditions. Robust growth was observed in all three effluents, with nejayote identified as the optimal medium. Physicochemical measurements conducted pre- and post-cultivation revealed notable reductions of pollutants in nejayote, including complete removal of nitrogen and phosphates, and an 85 % reduction in COD. Tequila vinasses exhibited promise with a 66 % reduction in nitrogen and a 70 % reduction in COD, while cheese-whey showed a 17 % reduction in phosphates. Regarding valuable compounds, nejayote yielded the highest pigment (1.62 mg·g-1) and phenolic compound (3.67 mg·g-1) content, while tequila vinasses had the highest protein content (16.83 %). The main highlight of this study is that C. vulgaris successfully grew in 100 % of the three effluents (without additional water or nutrients), demonstrating its potential for sustainable bioremediation and added-value compound production. When grown in 100 % of the effluents, they become a sustainable option since they don't require an input of fresh water and therefore do not contribute to water scarcity. These findings offer a practical solution for addressing environmental challenges in the food and beverage industries within a circular economy framework.

Biofixation of Air Emissions and Biomass Valorization-Evaluation of Microalgal Biotechnology.

This research appraised the simultaneous biofixation, that is not quite common in scientific literature, of carbon dioxide (CO2) and nitric oxides (NOx) by microalgae species Chlorella vulgaris, Haematococcus pluvialis, and Scenedesmus subspicatus. The experimental design was established by five treatments with gas concentrations between control-0.04% of CO2, 5 to 15% of CO2, and 30 to 100 ppm of NOx. Parameters such as pH, growth, productivity, lipids, protein, carbon/ nitrogen ratio, and astaxanthin were evaluated. For all species, the maximal growth and productivity were achieved with 5% of CO2 and 30 ppm of NOx. Regarding protein content, for all the three species, better results were obtained at higher concentrations of CO2 and NOx. These results prove the microalgae capacity for CO2 and NOx biofixation and reuse of biomass as a source of high value-added products, such as lipids, proteins, and astaxanthin. These findings support the indication of these species for flue gas treatment process and use in biorefineries systems.

Active indole-3-acetic acid biosynthesis by the bacterium Azospirillum brasilense cultured under a biogas atmosphere enables its beneficial association with microalgae.

AIMS: This study assessed, at the physiological and molecular levels, the effect of biogas on indole-3-acetic acid (IAA) biosynthesis by Azospirillum brasilense as well as the impact of this bacterium during CO2 fixation from biogas by Chlorella vulgaris and Scenedesmus obliquus. METHODS AND RESULTS: IpdC gene expression, IAA production and the growth of A. brasilense cultured under air (control) and biogas (treatment) were evaluated. The results demonstrated that A. brasilense had a better growth capacity and IAA production (105.7 ± 10.3 µg ml-1 ) when cultured under biogas composed of 25% CO2  + 75% methane (CH4 ) with respect to the control (72.4 ± 7.9 µg ml-1 ), although the ipdC gene expression level was low under the stressful condition generated by biogas. Moreover, this bacterium was able to induce a higher cell density and CO2 fixation rate from biogas by C. vulgaris (0.27 ± 0.08 g l-1 d-1 ) and S. obliquus (0.22 ± 0.08 g l-1 d-1 ). CONCLUSIONS: This study demonstrated that A. brasilense has the capacity to grow and actively maintain its main microalgal growth-promoting mechanism when cultured under biogas and positively influence CO2 fixation from the biogas of C. vulgaris and S. obliquus. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings broaden research in the field of Azospirillum-microalga interactions and the prevalence of Azospirillum in environmental and ecological topics in addition to supporting the uses of plant growth-promoting bacteria to enhance biotechnological strategies for biogas upgrading.

Removal and biotransformation of 4-nonylphenol by Arthrospira maxima and Chlorella vulgaris consortium.

4-Nonylphenol (4-NP) is an anthropogenic contaminant found in different environmental matrices that has an effect over the biotic and abiotic factors within the environment. Bioremediation by microorganisms can be used as a potential treatment to remove this pollutant. In this work, a consortium of two microorganisms, Arthrospira maxima and Chlorella vulgaris, was employed to remove 4-NP from water. The parameters analyzed included cell growth, removal of 4-NP, and 4-NP remnant in the biomass. In addition, the metabolites produced in the process by this consortium were identified. It was found that C. vulgaris is more resistant to 4-NP than A. maxima (cell growth inhibition by 4-NP of 99%). The consortium used in this study had an IC50 greater than any strain of microalgae or cyanobacteria reported for 4-NP removal (9.29 mg/L) and reduced up to 96% of 4-NP in water in the first 48 h of culture. It was also observed that there is a bio-transformation of 4-NP, comparable with the process carried out by another bacterium, in which three similar metabolites were found (4-(1-methyl-octyl)-4-hydroxy-cyclohex-2-enone, 4-nonyl-4-hydroxy-ciclohexa-2,5-dienone and 4-nonyl-4-hydroxy- ciclohex-2-enone) and one that is similar to plant metabolism (4-nonyl-(1-methyl,6,8-metoxy)-hydroxybenzene). These results indicate that microalgae and cyanobacteria consortium can be used to remove 4-NP from water.

Influence of organic matter and CO2 supply on bioremediation of heavy metals by Chlorella vulgaris and Scenedesmus almeriensis in a multimetallic matrix.

This research evaluated the influence of organic matter (OM) and CO2 addition on the bioremediation potential of two microalgae typically used for wastewater treatment: Chlorella vulgaris (CV) and Scenedesmus almeriensis (SA). The heavy metal (HM) removal efficiencies and biosorption capacities of both microalgae were determined in multimetallic solutions (As, B, Cu, Mn, and Zn) mimicking the highest pollutant conditions found in the Loa river (Northern Chile). The presence of OM decreased the total biosorption capacity, specially in As (from 2.2 to 0.0 mg/g for CV and from 2.3 to 1.7 mg/g for SA) and Cu (from 3.2 to 2.3 mg/g for CV and from 2.1 to 1.6 mg/g for SA), but its influence declined over time. CO2 addition decreased the total HM biosorption capacity for both microalgae species and inhibited CV growth. Finally, metal recovery using different eluents (HCl, NaOH, and CaCl2) was evaluated at two different concentrations. HCl 0.1 M provided the highest recovery efficiencies, which supported values over 85% of As, 92% of Cu, and ≈100% of Mn and Zn from SA. The presence of OM during the loaded stage resulted in a complete recovery of As, Cu, Mn, and Zn when using HCl 0.1 M as eluent.

Metabolic Pathway Analysis of Nitrogen and Phosphorus Uptake by the Consortium between C. vulgaris and P. aeruginosa.

Anthropogenic activities have increased the amount of urban wastewater discharged into natural aquatic reservoirs containing a high amount of nutrients such as phosphorus (Pi and PO 4 - 3 ), nitrogen (NH 3 and NO 3 - ) and organic contaminants. Most of the urban wastewater in Mexico do not receive any treatment to remove nutrients. Several studies have reported that an alternative to reduce those contaminants is using consortiums of microalgae and endogenous bacteria. In this research, a genome-scale biochemical reaction network is reconstructed for the co-culture between the microalga Chlorella vulgaris and the bacterium Pseudomonas aeruginosa. Metabolic Pathway Analysis (MPA), is applied to understand the metabolic capabilities of the co-culture and to elucidate the best conditions in removing nutrients. Theoretical yields for phosphorus removal under photoheterotrophic conditions are calculated, determining their values as 0.042 mmol of PO 4 - 3 per g DW of C. vulgaris, 19.43 mmol of phosphorus (Pi) per g DW of C. vulgaris and 4.90 mmol of phosphorus (Pi) per g DW of P. aeruginosa. Similarly, according to the genome-scale biochemical reaction network the theoretical yields for nitrogen removal are 10.3 mmol of NH 3 per g DW of P. aeruginosa and 7.19 mmol of NO 3 - per g DW of C. vulgaris. Thus, this research proves the metabolic capacity of these microorganisms in removing nutrients and their theoretical yields are calculated.

Production of Ethyl Esters by Direct Transesterification of Microalga Biomass Using Propane as Pressurized Fluid.

This work aimed to produce ethyl esters from Chlorella vulgaris microalgae biomass, using an immobilized enzymatic catalyst associated with pressurized fluid (propane) by direct transesterification. In order to optimize the ethyl conversion, different temperatures (46.7-68.1 °C) and pressures (59.5-200.5 bar) were applied a central composite design rotational (CCDR) obtaining the high conversion (74.39%) at 50 °C and 180 bar. The molar ratio also was investigated showing conversions ~ 90% using a molar ratio of 1:24 (oil:ethanol). From the best transesterification conditions, 50 °C, 180 bar, 20% enzymatic concentration, and 1:24 oil:ethanol molar ratio were obtained with success 98.9% conversion in 7 h of reaction. The enzyme reuse maintained its activity for three successive cycles. Thus, this simple process was effective to convert microalgal biomass into ethyl ester by direct transesterification and demonstrate high yields.

Biomass and lipid production from Chlorella vulgaris UTEX 26 cultivated in 2 m3 raceway ponds under semicontinuous mode during the spring season.

A Chlorella vulgaris UTEX 26 semicontinuous culture was implemented in 2000 L raceways with M medium during spring season at greenhouse conditions. Areal biomass productivities between 20 and 26 g m-2 d-1 were reached on the third day. The maximal areal lipid productivity obtained was 6.1 g m-2 d-1 and an increment in the saturated fatty acids (SFA) proportion (C14-C18) was favored in comparison with the fatty acids obtained with M medium in photobioreactors of 1 L and photoperiod light:darkness 12:12 h. After the eighth day of the culture or biomass concentrations above 0.25 g L-1, the microalgal cultures were prone to contamination by ciliates and amoebae, due to the sugars excreted by C. vulgaris UTEX 26. The periodical addition of NH4HCO3 to the microalgal culture maintained the ammonium concentration between 25 and 50 mg L-1, which contributed to diminish the contamination risks by protozoa.

Biodiesel synthesis from Chlorella vulgaris under effect of nitrogen limitation, intensity and quality light: estimation on the based fatty acids profiles.

Cultures under nitrogen limitation for Chlorella vulgaris were kept under different light quality (white, blue, yellow and violet) at 70 and 140 µE m-2 s-1; to evaluate the effect on fatty acids profiles and biodiesel quality. The results showed a maximum biomass and cell density at 140 µE m-2 s-1 of: white light (0.69 g L-1 and 6.5 × 106 cells mL-1, respectively) and blue light (0.65 g L-1 and 8.0 × 106 cells mL-1, respectively); compared to violet and yellow light. The chlorophyll concentration (µg mg-1 biomass dry weight) at 70 µE m-2 s-1 were in the order of light: white (25.61) > violet (17.10) > yellow (11.68) > blue (11.40) and, at 140 µE m-2 s-1 were: violet (23.64) > white (10.20) > yellow (9.66) > blue (7.99), suggesting the violet light stimulates the increase of chlorophyll a at higher intensity. The maximum lipid content (% w/w) were present under blue light (43.11), yellow (70.92) and violet (83.87) at 140 µE m-2 s-1. The different wavelengths did not have a negative effect on the quality of the biodiesel, however; violet light presented greater productivity and the indicators such as CFPP were related to the oxidative stability value and low PUFA content, leading biodiesel to good oxidative stability.

Chlorella vulgaris mixotrophic growth enhanced biomass productivity and reduced toxicity from agro-industrial by-products.

Algal wastewater remediation has become attractive for a couple of years now, however the effectiveness of genetic toxicity reducing of some by-products through microalgae are still not well reported. This study aimed to evaluate the growth, nutrients and toxicity removal of Chlorella vulgaris cultivated under autotrophic and mixotrophic conditions in three agro-industrial by-products. Mixotrophic culture using corn steep liquor showed higher cell concentration, specific growth rate, maximum cell productivity and biomass protein content when compared to cheese whey and vinasse. Nutrient removal results showed that C. vulgaris was able to completely remove corn steep liquor nutrients, while in cheese whey and vinasse culture this removal was not as efficient, observing remaining COD. This work evaluated for the first time the corn steep liquor and cheese whey genetic toxicity through Allium cepa seeds assay. These results demonstrate that corn steep liquor toxicity was totally eliminated by C. vulgaris cultivation, and cheese whey and vinasse toxicity were minimized. This study proves that the mixotrophic cultivation of C. vulgaris can increase cellular productivity, as well as it is a suitable and economic alternative to remove the toxicity from agroindustrial by-products.

Azospirillum brasilense Increases CO2 Fixation on Microalgae Scenedesmus obliquus, Chlorella vulgaris, and Chlamydomonas reinhardtii Cultured on High CO2 Concentrations.

Mutualism interactions of microalgae with other microorganisms are widely used in several biotechnological processes since symbiotic interaction improves biotechnological capabilities of the microorganisms involved. The interaction of the bacterium Azospirillum brasilense was assessed with three microalgae genus, Scenedesmus, Chlorella, and Chlamydomonas, during CO2 fixation under high CO2 concentrations. The results in this study have demonstrated that A. brasilense maintained a mutualistic interaction with the three microalgae assessed, supported by the metabolic exchange of indole-3-acetic acid (IAA) and tryptophan (Trp), respectively. Besides, CO2 fixation increased, as well as growth and cell compound accumulation, mainly carbohydrates, in each microalgae evaluated, interacting with the bacterium. Overall, these results propose the mutualism interaction of A. brasilense with microalgae for improving biotechnological processes based on microalgae as CO2 capture and their bio-refinery capacity.

Bioelectrogenesis with microbial fuel cells (MFCs) using the microalga Chlorella vulgaris and bacterial communities

Background: Microbial Fuel Cell (MFC) technology is used in various applications such as wastewater treatment with the production of electrical energy. The objective of this study was to estimate the biodepuration of oils and fats, the elimination of blue dye brl and bioelectro-characterization in MFCs with Chlorella vulgaris and bacterial community. Results: The operation of MFCs at 32 d showed an increase in bioelectrogenic activity (from 23.17 to 327.67 mW/m2 ) and in the potential (from 200 to 954 mV), with biodepuration of fats and oils (95%) in the microalgal cathode, and a removal of the chemical oxygen demand COD (anode, 71%, cathode, 78.6%) and the blue dye brl (73%) at the anode, here biofilms were formed by the bacterial community consisting of Actinobacteria and Deltaproteobacteria. Conclusions: These findings suggest that MFCs with C. vulgaris and bacterial community have a simultaneous efficiency in the production of bioelectricity and bioremediation processes, becoming an important source of bioenergy in the future.

Evaluation of nutrients removal (NO3-N, NH3-N and PO4-P) with Chlorella vulgaris, Pseudomonas putida, Bacillus cereus and a consortium of these microorganisms in the treatment of wastewater effluents.

In this research removal of NH3-N, NO3-N and PO4-P nutrients from municipal wastewater was studied, using Chlorella vulgaris, Pseudomonas putida, Bacillus cereus and an artificial consortium of them. The objective is to analyze the performance of these microorganisms and their consortium, which has not been previously studied for nutrient removal in municipal wastewater. A model wastewater was prepared simulating the physicochemical characteristics found at the wastewater plant in Chapala, Mexico. Experiments were carried out without adding an external carbon source. Results indicate that nutrient removal with Chlorella vulgaris was the most efficient with a removal of 24.03% of NO3-N, 80.62% of NH3-N and 4.30% of PO4-P. With Bacillus cereus the results were 8.40% of NO3-N, 28.80% of NH3-N and 3.80% of PO4-P. The removals with Pseudomonas putida were 2.50% of NO3-N, 41.80 of NH3-N and 4.30% of PO4-P. The consortium of Chlorella vulgaris-Bacillus cereus-Pseudomonas putida removed 29.40% of NO3-N, 4.2% of NH3-N and 8.4% of PO4-P. The highest biomass production was with Bacillus cereus (450 mg/l) followed by Pseudomonas putida (444 mg/l), the consortium (205 mg/l) and Chlorella vulgaris (88.9 mg/l). This study highlights the utility of these microorganisms for nutrient removal in wastewater treatments.

Green production of microalgae-based silver chloride nanoparticles with antimicrobial activity against pathogenic bacteria.

Silver nanoparticles are powerful antimicrobial agents. Here, the synthesis of silver chloride nanoparticles (AgCl-NPs) was consistently evidenced from a commercially valuable microalgae species, Chlorella vulgaris. Incubation of C. vulgaris conditioned medium with AgNO3 resulted in a medium color change to yellow/brown (with UV-vis absorbance at 415nm), indicative of silver nanoparticle formation. Energy-dispersive X-ray spectroscopy (EDS) of purified nanoparticles confirmed the presence of both silver and chlorine atoms, and X-ray diffraction (XRD) showed the typical pattern of cubic crystalline AgCl-NPs. Transmission electron microscopy (TEM) showed that most particles (65%) were spherical, with average diameter of 9.8±5.7nm. Fourier transform infrared spectroscopy (FTIR) of purified nanoparticle fractions suggested that proteins are the main molecular entities involved in AgCl-NP formation and stabilization. AgCl-NPs (from 10µg/mL) decreased by 98% the growth of Gram-positive Staphylococcus aureus and Gram-negative Klebsiella pneumoniae bacterial pathogens, and had a dose-dependent effect on cell viability, which was measured by automated image-based high content screening (HCS). Ultrastructural analysis of treated bacteria by TEM revealed the abnormal arrangement of the chromosomal DNA. Our findings strongly indicated that the AgCl-NPs from C. vulgaris conditioned medium is a promising 'green' alternative for biomedical application as antimicrobials.

A novel culture medium designed for the simultaneous enhancement of biomass and lipid production by Chlorella vulgaris UTEX 26.

A novel culture medium to enhance the biomass and lipid production simultaneously by Chlorella vulgaris UTEX 26 was designed in three stages of optimization. Initially, a culture medium was inferred applying the response surface method to adjust six factors [NaNO3, NH4HCO3, MgSO4·7H2O, KH2PO4, K2HPO4 and (NH4)2HPO4], which were selected on the basement of BBM (Bold's Basal Medium) and HAMGM (Highly Assimilable Minimal Growth Medium) culture media. Afterwards, the nitrogen source compound was optimized to reduce both, ammonium and nitrate concentrations. As result of the optimization process, the proposed culture medium improved 40% the biomass (0.73gL(-1)) compared with the BBM medium and 85% the lipid concentration (281mgL(-1)), with respect to HAMGM medium. Some culture media components concentrations were reduced up to 50%. Gas chromatography analysis revealed that C16:0, C18:0, C18:1, C18:2 and C18:3 were the major fatty acids produced by C. vulgaris UTEX 26.

NMR techniques for determination of lipid content in microalgal biomass and their use in monitoring the cultivation with biodiesel potential.

In the present investigation, the application of NMR spectroscopic techniques was extensively used with an objective to explore the biodiesel potential of biomass cultivated on a lab scale using strains of Chlorella vulgaris and Scenedesmus ecornis. The effect of variation in the composition of culturing medium on the neutral and polar lipids productivity, and fatty acid profile of solvent extracts of microalgae biomass was studied. Determination of unsaturated fatty acid composition (C18:N = 1-3, ω3 C20:5, ω3 C22:6), polyunsaturated fatty esters (PUFEs), saturated fatty acids (SFAs), unsaturated fatty acids (UFAs), free fatty acids (FFAs), and iodine value were achieved from a single (1)H NMR spectral analysis. The results were validated by (13)C NMR and GC-MS analyses. It was demonstrated that newly developed methods based on (1)H and (13)C NMR techniques are direct, rapid, and convenient for monitoring the microalgae cultivation process for enhancement of lipid productivity and their quality aspects in the solvent extracts of microalgal biomasses without any sample treatment and prior separation compared to other methods. The fatty acid composition of algae extracts was found to be similar to vegetable and fish oils, mostly rich in C16:0, C18:N (N = 0 to 3), and n-3 omega polyunsaturated fatty acids (PUFAs). The lipid content, particularly neutral lipids, as well as most of the quality parameters were found to be medium specific by both the strains. The newly developed methods based on NMR and ultrasonic procedure developed for efficient extraction of neutral lipids are cost economic and can be an effective aid for rapid screening of algae strains for modulation of lipid productivity with desired biodiesel quality and value-added products including fatty acid profile.

Accumulation of intra-cellular polyphosphate in Chlorella vulgaris cells is related to indole-3-acetic acid produced by Azospirillum brasilense.

Accumulation of intra-cellular phosphate, as polyphosphate, was measured when the microalga Chlorella vulgaris was immobilized in alginate with either of two wild-type strains of the microalgae growth-promoting bacterium Azospirillum brasilense or their corresponding IAA-attenuated mutants. Wild type strains of A. brasilense induced higher amounts of intra-cellular phosphate in Chlorella than their respective mutants. Calculations comparing intra-cellular phosphate accumulation by culture or net accumulation by the cell and the amount of IAA that was produced by each of these strains revealed that higher IAA was linked to higher accumulations of intra-cellular phosphate. Application of four levels of exogenous IAA reported for A. brasilense and their IAA-attenuated mutants to cultures of C. vulgaris enhanced accumulation of intra-cellular phosphate; the higher the content of IAA per culture or per single cell, the higher was the amount of accumulated phosphate. When an IAA-attenuated mutant was complemented with exogenous IAA, accumulation of intra-cellular phosphate at the culture level was even higher than phosphate accumulation with the respective wild type strains. When calculating the net accumulation of intra-cellular phosphate in the complementation experiment, net intra-cellular phosphate induced by the IAA-attenuated mutant was completely restored and was similar to the wild strains. We propose that IAA produced by A. brasilense is linked to polyphosphate accumulation in C. vulgaris.

Bioremoval of the azo dye Congo Red by the microalga Chlorella vulgaris.

Discharge of dye-containing wastewater by the textile industry can adversely affect aquatic ecosystems and human health. Bioremoval is an alternative to industrial processes for detoxifying water contaminated with dyes. In this work, active and inactive biomass of the microalga Chlorella vulgaris was assayed for the ability to remove Congo Red (CR) dye from aqueous solutions. Through biosorption and biodegradation processes, Chlorella vulgaris was able to remove 83 and 58 % of dye at concentrations of 5 and 25 mg L(-1), respectively. The maximum adsorption capacity at equilibrium was 200 mg g(-1). The Langmuir model best described the experimental equilibrium data. The acute toxicity test (48 h) with two species of cladocerans indicated that the toxicity of the dye in the effluent was significantly decreased compared to the initial concentrations in the influent. Daphnia magna was the species less sensitive to dye (EC50 = 17.0 mg L(-1)), followed by Ceriodaphnia dubia (EC50 = 3.32 mg L(-1)). These results show that Chlorella vulgaris significantly reduced the dye concentration and toxicity. Therefore, this method may be a viable option for the treatment of this type of effluent.

Effects of different biomass drying and lipid extraction methods on algal lipid yield, fatty acid profile, and biodiesel quality.

Three lipid extraction methods of hexane Soxhlet (Sox-Hex), Halim (HIP), and Bligh and Dyer (BD) were applied on freeze-dried (FD) and oven-dried (OD) Chlorella vulgaris biomass to evaluate their effects on lipid yield, fatty acid profile, and algal biodiesel quality. Among these three methods, HIP was the preferred one for C. vulgaris lipid recovery considering both extraction efficiency and solvent toxicity. It had the highest lipid yields of 20.0 and 22.0% on FD and OD biomass, respectively, with corresponding neutral lipid yields of 14.8 and 12.7%. The lipid profiling analysis showed that palmitic, oleic, linoleic, and α-linolenic acids were the major fatty acids in the algal lipids, and there were no significant differences on the amount of these acids between different drying and extraction methods. Correlative models applied to the fatty acid profiles concluded that high contents of palmitic and oleic acids in algal lipids contributed to balancing the ratio of saturated and unsaturated fatty acids and led to a high-quality algal biodiesel.