Auxenochlorella Protothecoides Research Articles

Xylose, glucose and acetate as feedstock for three microalgal species cultivated in heterotrophy

Microalgae are known as good producers of high-added value bioproducts useful in many applications such as pharmaceuticals, nutrition or biofuel production. In contrast to phototrophy, heterotrophy emerges as a promising strategy for algal biomass production due to high cell densities, controlled conditions and reduced space requirement. Hemicellulose is the second most abundant material in land plants. Its hydrolysis liberates xylose, glucose, acetate. Our study focuses on three microalgal species, Galdieria sulphuraria, Euglena gracilis, and Auxenochlorella protothecoides, cultivated under heterotrophy with the above-mentioned carbon sources, supplemented alone or in combination. Growth parameters and biomass analysis revealed distinctive characteristics. G. sulphuraria, despite a modest fatty acid content (5–15 % w/w), displayed potential for hemicellulose valorization, demonstrating high biomass yield using xylose as a sole carbon source (approx. 0.5 gDW gxylose−1) and high saturated fatty acid (SFA) content (45–63 %). In our cultivation conditions, E. gracilis only assimilated acetate with low fatty acid content (approx. 6 % w/w), but high SFA content (60–77 %) and high paramylon content (47 % w/w), convertible to wax-esters under anaerobiosis. A. protothecoides exhibited biomass yields of 0.42–0.54 gDW gsubstrate−1 depending on the carbon substrate supplied but maintained constant fatty acid content (16–18 % w/w) in the presence of all substrates except xylose. Surprisingly, despite an inability to grow with xylose alone, sugar depletion analysis indicated decreasing xylose concentration when other carbon sources were present in the cultivation medium for this alga. This comparative study discusses the strengths and weaknesses of each strain, providing insights into their potential when grown on hemicellulose carbon sources.

Soy whey and brewer’s spent grain hydrolysates wholly replace conventional medium for microalgae growth: Process performance and economic considerations

To enhance circularity in heterotrophic microalgal bioprocesses, this study completely substituted glucose and Bold’s basal medium (BBM) with brewer’s spent grain (BSG) and soy whey (SW) hydrolysates. Mild acid hydrolysis conditions of BSG (0.2 M H2SO4, 130 °C, 36 min) and SW (0.1 M HCl, 95 °C, 30 min) were optimised for glucose release, and their hydrolysates were optimally mixed (15 % SW-85 % BSG) to obtain a medium that best supported Auxenochlorella protothecoides growth. Maximum biomass production (Xmax) and productivity (PXmax) obtained in the hydrolysate medium containing 50.75 g/L endogenous glucose (Xmax: 22.17 g/L; PXmax: 7.06 g/L/day) were comparable to that in BBM containing 50.44 g/L exogenous glucose (Xmax: 20.02 g/L; PXmax: 6.34 g/L/day). Moreover, estimated hydrolysate medium production costs were within an order of magnitude to BBM. Overall, the integrated approach of tailored hydrolytic treatments and complementary side-streams presents a promising technical and economic feasibility, with applications extending beyond A. protothecoides.

Enhancing protein extraction from heterotrophic Auxenochlorella protothecoides microalgae through emerging cell disruption technologies combined with incubation

This study evaluated pulsed electric fields (PEF) and ultrasonication (US) combined with incubation to enhance cell disruption and protein extraction from Auxenochlorella protothecoides, comparing them to conventional high-pressure homogenization (HPH). A 5 h incubation enhanced protein yield by 79.4 % for PEF- and 27.2 % for US-treated samples. Extending the incubation to 24 h resulted in a total yield increase of 122 % for PEF (0.25 ± 0.03 kgEP kgTP−1) and 51.9 % for US (0.20 ± 0.02 kgEP−1 kgTP−1). Autofermentation in untreated cells after 24 h resulted in protein release with lower yields than all other treated and incubated samples. While HPH had the highest protein yield (0.58 ± 0.04 kgEP kgTP−1), PEF-incubation after 5 h (56.6 ± 5.3 MJ kgEP−1) and 24 h (49.5 ± 3.7 MJ kgEP−1) were 1.5 and 1.7-times more energy-efficient than HPH (82.9 ± 7.8 MJ kgEP−1). PEF combined incubation is an energy-efficient and targeted protein extraction method in heterotrophic A. protothecoides biorefinery.

Acclimated green microalgae consortium to treat sewage in an alternative urban WWTP in a coastal area of Central Italy

This study exposed a microalgal consortium formed by Auxenochlorella protothecoides, Tetradesmus obliquus, and Chlamydomonas reinhardtii to six mixed wastewater media containing different proportions of primary (P) or secondary (S) effluents diluted in centrate (C). Algae could grow at centrate concentrations up to 50 %, showing no significant differences between effluents. After acclimation, microalgae cultivated in 50%P-50%C and 50%S-50%C grew at a rate similar to that of control cultures (0.59–0.66 d−1). These results suggest that the consortium acclimated to both sewage streams by modulating the proportion of the species and their metabolism. Acclimation also altered the photosynthetic activity of wastewater-grown samples compared to the control, probably due to partial photoinhibition, changes in consortium composition, and changes in metabolic activity. No major differences were observed between the two streams with respect to biochemical composition, biomass yield, or bioremediation capacity of the cultivated algae but algae grown in the secondary effluent showed qualitatively higher exopolysaccharides (EPS) production than algae grown in primary. Regarding wastewater remediation, microalgae grown in both WW media showed proficient nutrient removal efficiencies (close to 100 %); however, the final pH value (close to 11) would be controversial if the system were upscaled as it is over the legal limit and would cause phosphorus precipitation, so that CO2 addition would be required. The theoretical scale-up of the microalgae system could achieve water treatment costs of 0.109 €·m−3, which was significantly lower than the costs of typical activated sludge systems.

Two-Dimensional Thin-Layer Chromatography as an Accessible, Low-Cost Tool for Lipid-Class Profile Screening

The interest in lipid composition profiling is significantly increasing as research reveals the immense importance of lipids in medicine, plant science, food and agriculture. However, lipidomic analysis requires high-end specialty equipment. We used two-dimensional thin-layer chromatography (2D-TLC) as a readily available, low-cost tool for basic lipidomic profiling of lipid classes in algal samples in the models Chlamydomonas reinhardtii, Auxenochlorella protothecoides, and Euglena gracilis, validating lipid class identification using an LC-MS/MS analysis. Algal lipid extracts were separated on a 2D-TLC plate, and TLC analysis was followed by scraping individual TLC spots off the plate, and a subsequent liquid chromatography separation and tandem mass spectrometry (LC-MS/MS) analysis. For comparison, crude lipid extracts were also injected directly to the LC-MS/MS system. Lipid class annotation was achieved by a combination of accurate mass, retention time information, neutral loss and fragment ion analysis by MS2Analyzer, and by matching spectra to LipidBlast MS/MS library. Overall, we were able to identify 15 lipid classes, and to adequately profile the lipid classes in all three organisms. This TLC method is thus suggested as an accessible tool for lipid class profiling of algal, plant, and food lipids, alike, when a rapid and simple analysis is required, e.g., for screening purposes.

Growth and Cell Size of Microalga Auxenochlorella protothecoides AS-1 under Different Trophic Modes.

Certain microalgal species can grow with different trophic strategies depending on the availability of nutrient resources. They can use the energy from light or an organic substrate, or both, and can therefore be called autotrophs, heterotrophs, or mixotrophs. We recently isolated a microalgal strain from the microplastic biofilm, which was identified as Auxenochlorella protothecoides, AS-1. Strain AS-1 grew rapidly in bacterial culture media and exhibited different growth rates and cell sizes under different trophic conditions. We compared the growth performance of AS-1 under the three different trophic modes. AS-1 reached a high biomass (>4 g/L) in 6 days under mixotrophic growth conditions with a few organic carbons as a substrate. In contrast, poor autotrophic growth was observed for AS-1. Different cell sizes, including daughter and mother cells, were observed under the different growth modes. We applied a Coulter Counter to measure the size distribution patterns of AS-1 under different trophic modes. We showed that the cell size distribution of AS-1 was affected by different growth modes. Compared to the auto-, hetero- and mixotrophic modes, AS-1 achieved higher biomass productivity by increasing cell number and cell size in the presence of organic substrate. The mechanisms and advantages of having more mother cells with organic substrates are still unclear and warrant further investigations. The work here provides the growth information of a newly isolated A. protothecoides AS-1 which will be beneficial to future downstream applications.

Removal efficiencies of seven frequently detected antibiotics and related physiological responses in three microalgae species

The main objective of this study is to investigate the effect of mixtures of seven widely used human antibiotics (ciprofloxacin, clarithromycin, erythromycin, metronidazole, ofloxacin, sulfamethoxazole, and trimethoprim) on the growth, pH, pigment production, and antibiotics removal of three microalgal species (Auxenochlorella protothecoides, Tetradesmus obliquus, and Chlamydomonas acidophila). Batch assays were conducted with media with antibiotic mixtures at 10, 50, and 100 μg L−1 for each antibiotic. The three microalgae species effectively removed the antibiotics without any growth inhibition, even when exposed to the highest antibiotic concentrations. Biosorption was reported as the primary mechanism for ciprofloxacin, clarithromycin, metronidazole, and ofloxacin, with up to 70% removal, especially in A. protothecoides and C. acidophila. A. protothecoides, a species never investigated for antibiotic removal, was the only microalgae exhibiting bioaccumulation and biodegradation of specific antibiotics, including sulfamethoxazole. Furthermore, in media with the highest antibiotic concentration, all three species exhibited increased chlorophyll (up to 37%) and carotenoid (up to 32%) production, accompanied by a pH decrease of 3 units. Generally, in the present study, it has been observed that physiological responses and the removal of antibiotics by microalgae are interlinked and contingent on the antibiotic levels and types.

Biotechnological product potential of Auxenochlorella protothecoides including biologically active compounds (BACs) under nitrogen stress conditions.

Nitrogen stress can influence microalgae's growth characteristics, and microalgae grown in nitrogen-deficient conditions may produce higher or lower levels of biotechnological products as a result of metabolic changes. In photoautotrophic and heterotrophic cultures, nitrogen limitation has been proven effective in promoting lipid accumulation. In spite of this, no study has demonstrated a significant correlation between lipid content and other biotechnological products such as bioactive compounds (BACs).This research examines a strategy for lipid accumulation as well as the potential production of BACs with antibacterial properties in parallel with that strategy. This concept involved the treatment of the microalga Auxenochlorella protothecoides with low and high concentrations of ammonium (NH4+). This particular experiment reached a maximum lipid content of 59.5% using a 0.8 mM NH4+ concentration, resulting in the yellowing of the chlorophyll levels. Agar diffusion assays were conducted to determine the antibacterial activity of different extracts derived from the biomass when stressed with different levels of nitrogen.Algal extracts prepared by a variety of solvents showed different levels of antibacterial activity against representative strains of both gram-negative (Escherichia coli) and gram-positive (Staphylococcus aureus) bacteria. Among the extracts tested, 500mg/L ethyl acetate extract had the greatest antibacterial activity against Escherichia coli. In order to identify the components responsible for the extract's antibacterial activity, fatty acid methyl ester (FAME) analysis was performed. It has been suggested that the lipid fraction may be a valuable indicator of these activities since some lipid components are known to possess antimicrobial properties. In this regard, it was found that the amount of polyunsaturated fatty acid (PUFA) significantly decreased by 53.4% under the conditions with the highest antibacterial activity observed.

Production and characterization of exopolysaccharides from salinity-induced Auxenochlorella protothecoides and the analysis of anti-inflammatory activity

The set scenario of this work was to investigate the production, physicochemical characteristics, and anti-inflammatory activities of exopolysaccharides from salinity-induced Auxenochlorella protothecoides. The results demonstrated that 10 ‰ salinity manipulation endowed preferable exopolysaccharide production by A. protothecoides. Under this salinity stress, ACPEPS1A and ACPEPS2A were purified from exopolysaccharide production by anion chromatography and molecular exclusion chromatography. ACPEPS1A exhibited a molecular weight (Mw) of 132 kDa and mainly consisted of galactose. ACPEPS2A was a heteropolysaccharide with an Mw of 170 kDa and the main monosaccharides of galactose and rhamnose with separate molar percents of 42.41 % and 35.29 %, respectively. FTIR, 1H and 13C NMR supported that monosaccharide components of ACPEPS1A and ACPEPS2A possessed both α- and β-configuration pyranose rings. Further evidence indicated that ACPEPS1A and ACPEPS2A could effectively inhibit the inflammatory response in lipopolysaccharide (LPS) induced RAW264.7 cells by quenching inflammatory factor levels such as ROS, iNOS, TNF-α, and IL-6. The potential anti-inflammatory possibilities were that the monosaccharides of ACPEPS1A and ACPEPS2A possessed higher affinity with receptors on the macrophage surface than LPS and hampered LPS-induced inflammation. The findings of this work would favor innovative applications of exopolysaccharides from microalgae in complementary medicines or functional foods.

Expression profile of genes associated with the accumulation of triacylglycerol (TAG) in Auxenochlorella protothecoides KP7 under alkaline pH and nitrogen starvation conditions

Identification of target genes related to lipid accumulation in microalgae can increase triacylglycerol production without affecting biomass production through genetic manipulation. This study is part of our previous study and aimed to determine the effects of nitrogen starvation, alkaline pH (pH 10), and a combination of these factors on the transcriptional profiles of eight genes potentially related to triacylglycerol accumulation in Auxenochlorella protothecoides KP7. As shown previously, triacylglycerol content in Auxenochlorella protothecoides KP7 reached the maximum level under combined stress conditions. In this study, the highest upregulation for the gene encoding phosphoenolpyruvate carboxylase (PEPC) was observed under both nitrogen starvation and combined stress. Nitrogen starvation alone resulted in a significant increase in the expression level of all genes except NADP-malic enzyme (MEchl), which is localized in the chloroplast. Compared with nitrogen starvation alone, the upregulation of pyruvate dehydrogenase complex E2 component (PDHC-E2), glycerol-3-phosphate dehydrogenase (GPDH), diacylglycerol acyltransferase 1 (DGAT1), and MEchl genes further increased under the combined stress. These results suggest that genes encoding the enzymes DGAT1, GPDH, ME, PEPC, and PDHC-E2 are associated with triacylglycerol accumulation and could be used as target genes for genetic modification of Auxenochlorella protothecoides and possibly other microalgal species.

The Physiological Effects of Organic Solvents, Plant Growth Regulators, and Fungicide on Auxenochlorella protothecoides

The Physiological Effects of Organic Solvents, Plant Growth Regulators, and Fungicide on Auxenochlorella protothecoides

Direct transesterification of microalgae after pulsed electric field treatment

AbstractBACKGROUNDLipid extraction is a major bottleneck for the commercialization of microalgae due to energy costs involved during solvent recycling. Direct transesterification offers the possibility to bypass the extraction step by immediately converting the lipids to fatty acid methyl esters (FAMEs). In this study, the efficiency of direct transesterification after pulsed electric field (PEF) treatment was evaluated. Freshly harvested Auxenochlorella protothecoides (A. protothecoides), cultivated either autotrophically or mixotrophically, was subjected to PEF. Two treatment energies were tested, 0.25 and 1.5 MJ kgdw−1, and results were compared with those for conventional two‐step transesterification.RESULTSFor autotrophically grown A. protothecoides, the percentage of the total FAMEs recovered from untreated biomass and microalgae treated with 0.25 MJ kgdw−1 was 30% for both cases, while for 1.5 MJ kgdw−1 it was 65%. A 24 h incubation step between PEF treatment and direct transesterification significantly improved the results. Untreated biomass remained stable with 30% of FAMEs, while with both treatment energies a 97% FAME recovery was achieved. However, for mixotrophic A. protothecoides the process was not as effective. Approximately 30% of FAMEs were recovered for all three conditions immediately after PEF with only a marginal increase after incubation. The reason for this different behavior of the two cultivation modes is unknown and under investigation.CONCLUSIONSOverall, the synergy between PEF and direct transesterification was proven to have potential, in particular for autotrophic microalgae. Its implementation and further optimization in a biorefinery therefore merit further attention. © 2022 The Authors. Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI).

An In Silico Framework to Mine Bioactive Peptides from Annotated Proteomes: A Case Study on Pancreatic Alpha Amylase Inhibitory Peptides from Algae and Cyanobacteria.

Bioactive peptides may exert beneficial activities in living organisms such as the regulation of glucose metabolism through the inhibition of alpha amylases. Algae and cyanobacteria are gaining a growing interest for their health-promoting properties, and possible effects on glucose metabolism have been described, although the underlying mechanisms need clarification. This study proposes a computer-driven workflow for a proteome-wide mining of alpha amylase inhibitory peptides from the proteome of Chlorella vulgaris, Auxenochlorella protothecoides and Aphanizomenon flos-aquae. Overall, this work presents an innovative and versatile approach to support the identification of bioactive peptides in annotated proteomes. The study: (i) highlighted the presence of alpha amylase inhibitory peptides within the proteomes under investigation (including ELS, which is among the most potent inhibitory tripeptides identified so far); (ii) mechanistically investigated the possible mechanisms of action; and (iii) prioritized further dedicated investigation on the proteome of C. vulgaris and A. flos-aquae, and on CSSL and PGG sequences.

Improvement of Lutein Production in Auxenochlorella protothecoides Using Its Genome-Scale Metabolic Model and a System-Oriented Approach.

Lutein is a valuable metabolite widely used in the food, pharmaceutical, cosmetic, and aquaculture industries. Marigold flowers are the most common source of commercial lutein, but cultivation area, weather conditions, and high manpower costs are among the disadvantages of lutein production from marigold flowers. Microalgae are an excellent alternative to plant sources of lutein as they do not have the limitations of plant extraction. Auxenochlorella protothecoides is a promising candidate for commercial production of lutein. In the present research, a genome-scale metabolic model was applied to introduce some strategies to improve lutein production in A. protothecoides. The effective reactions to improve lutein production were determined based on analysis of multiple optimal solutions. The enzymatic regulators of candidate reactions were identified using the BRENDA database. The effect of 13 activators was investigated experimentally. Our results showed that sodium citrate has the greatest effect on lutein production, so it was introduced as the most effective compound for increasing lutein production by A. protothecoides.

Auxenochlorella protothecoides populations adapted to low phosphate conditions accumulated more non-phosphorus glycerolipids and biomass than wild type progenitors

Biodiesel produced by microalgae has great potential as a portable energy that can replace traditional hydrocarbon sources. One limitation of scaling up algal cultivation is the availability of macronutrients, particularly inorganic phosphate (Pi), which is a finite and dwindling resource. Here, Auxenochlorella protothecoides was adapted to low Pi conditions by continuous cultivation in growth media containing 100 times less Pi (17.15 µM PO4) than replete media for ∼ 40 generations. The low Pi-adapted A. protothecoides populations showed significantly higher growth rates, compared to wild type (WT) (natural, non-mutated) progenitor populations in batch experiments, with average maximum growth rates of 0.72 d−1 and 0.54 d−1, respectively. Total lipid profiling of the adapting A. protothecoides populations indicated a shift to non-phosphorus glycerolipids, based on UPLC/MS analyzes. The proportions of monogalactosyldiacylglycerol (MGDG) and sulfoquinovosyldiacyglycerol (SQDG) fluctuated during adaptation, accumulating 305% and 317% of the WT levels respectively by the final sampling. Time-course transcriptome profiling of A. protothecoides across all adaptation stages revealed initial increases in transcript levels, followed by global decreased expression. The short-term transcriptomic changes, prior to ∼ 11 generations, were associated with major metabolic pathways. The long-term changes indicated increased fatty acid turnover and a decrease in photosynthesis-related gene expression. Transcripts predicted to encode alternative oxidase and pyrophosphate-dependent phosphofructokinase fluctuated during adaptation. The selection of A. protothecoides under low Pi conditions resulted in a microalga variant that after only ∼40 generations utilized Pi more efficiently for growth than its wild type progenitor population, while also producing 1.22 times more biomass. The adaptive processes described herein produced commercially relevant strain material and provide avenues for future, targeted engineering of molecular pathways for increased Pi use efficiency in similar organisms.

Post-incubation pH Impacts the Lipid Extraction Assisted by Pulsed Electric Fields from Wet Biomass of Auxenochlorella protothecoides

Pulsed electric field (PEF) treatment is a promising technology for efficient lipid extraction from microalgae. This study focusses on under-investigated processing parameters, such as media pH, pulse application, and incubation protocols. The lipid yield and electroporation level of PEF-treated Auxenochlorella protothecoides were determined at a medium pH of 3.0 and 5.0 under variation of the pre- or post-PEF incubation time and for split-dose treatments. Low energetic PEF treatments with 40 kV/cm and 1 μs pulses at 9.6 and 19.2 kJ/L were performed either in batch mode or in continuous flow. Post-PEF incubation significantly increased the shared electroporated cells (>60%) in medium pH 3.0, while no change was observed at pH 5.0. Split-dose PEF treatments at pH 5.0 caused significantly higher electroporation levels and lipid extraction yields than equivalent single-dose treatments. Results have shown that medium pH is critical in the final electroporation and lipid extraction yields of A. protothecoides and therefore should be considered in further studies.

Extracellular Metabolites of Heterotrophic Auxenochlorella protothecoides: A New Source of Bio-Stimulants for Higher Plants.

The biodiversity of microalgal species is enormous, and their versatile metabolism produces a wide diversity of compounds that can be used in food, healthcare, and other applications. Microalgae are also a potential source of bio-stimulants that enhance nutrition efficiency, abiotic stress tolerance, and/or crop quality traits. In this study, the extracellular metabolites of Auxenochlorella protothecoides (EAp) were prepared using three different culture strategies, and their effects on plant growth were examined. Furthermore, the composition of EAp was analyzed by GC-MS. The elongation of lateral roots and the cold-tolerance of Arabidopsis thaliana and Nicotiana benthamiana were promoted by EAp. Moreover, EAp from high-cell-density fermentation stimulated the growth of the leafy vegetables Brassica rapa and Lactuca sativa at dilutions as high as 500- and 1000-fold. Three major groups of compounds were identified by GC-MS, including organic acids or organic acid esters, phenols, and saccharides. Some of these compounds have known plant–stimulating effects, while the rest requires further investigation in the future. Our study demonstrates that EAp is a potential bio-stimulant, while also providing an environmentally friendly and economical microalgae fermentation process.

New concept of biodiesel production using food waste digestate powder: Co-culturing algae-activated sludge symbiotic system in low N and P paper mill wastewater

This paper aims to demonstrate an innovative process for the conversion of food waste digestate (FWD) powder into biofuel. The effects of different doses of FWD are investigated on microalgae-activated sludge (MAS) in treating pulp and paper mill wastewater (PPW) which generally contains insufficient nitrogen and phosphorus. FWD was added to adjust the initial N:P molar ratio in MAS at various levels (8:1 to 15:1). The highest Auxenochlorella protothecoides biomass achieved was 1.67 gL−1 at a 13.45:1 N/P molar ratio of PPW. After 10 days of cultivation, Auxenochlorella protothecoides-activated sludge system removed 91.7 %, 74.6 %, and 91.5 % of total nitrogen, phosphorus, and sCOD respectively at D0.836 gL−1 DD. The highest lipid productivity was reported as 41.27 ± 2.43 mg L−1 day−1. Fatty acid methyl ester (FAME) analysis showed the presence of an appreciable percentage of balanced saturated and unsaturated fatty acids i.e. palmitic, oleic, and linoleic acid, rendering its potential as a feedstock for biodiesel production. Activated sludge induced flocculation of Auxenochlorella protothecoides was measured. The whole process establishes an effective means of circular economy, where the secondary source of recyclable nutrients i.e. FWD will be used as a source of N and P in PPW to obtain algal biodiesel from a negative value industrial wastewater.

Enhancement of black and odorous water treatment coupled with accelerated lipid production by microalgae exposed to 12C6+ heavy-ion beam irradiation

In this study, Auxenochlorella protothecoides (AP-CK) was selected due to its reported high growth potential in sterilized black and odorous water (SBOW). In order to improve the resource utilization level of microalgae for wastewater treatment, AP-CK was mutated using 12C6+ heavy-ion beam irradiation, and a high lipid-containing mutant (AP-34#) was isolated and further evaluated to treat original black and odorous water (OBOW). Compared with the wild type, the maximum removal rates of COD, NH4+-N and TP of the mutant increased by 8.12 ± 0.33%, 10.43 ± 0.54% and 11.97 ± 0.16%, respectively, while maximum dissolved oxygen content increased from 0 to 4.36 ± 0.25 mg/L. Besides, the mutant lipid yield increased by 115.87 ± 3.22% over the wild type in OBOW. The fatty acid profile of AP-34# grown in SBOW and OBOW showed higher proportion of saturated fatty acids (C16:0 and C18:0) and valuable polyunsaturated fatty acids (mainly C20:5n3 and C22:6n3) which are more suitable for biodiesel production and value-added products, respectively. This work provides a new perspective on improving the characteristics of microalgae and an innovative approach for resource-based microalgae wastewater treatment through bioremediation of black and odorous water.

Fe3O4-PEI Nanocomposites for Magnetic Harvesting of Chlorella vulgaris, Chlorella ellipsoidea, Microcystis aeruginosa, and Auxenochlorella protothecoides.

Magnetic separation of microalgae using magnetite is a promising harvesting method as it is fast, reliable, low cost, energy-efficient, and environmentally friendly. In the present work, magnetic harvesting of three green algae (Chlorella vulgaris, Chlorella ellipsoidea, and Auxenochlorella protothecoides) and one cyanobacteria (Microcystis aeruginosa) has been studied. The biomass was flushed with clean air using a 0.22 μm filter and fed CO2 for accelerated growth and faster reach of the exponential growth phase. The microalgae were harvested with magnetite nanoparticles. The nanoparticles were prepared by controlled co-precipitation of Fe2+ and Fe3+ cations in ammonia at room temperature. Subsequently, the prepared Fe3O4 nanoparticles were coated with polyethyleneimine (PEI). The prepared materials were characterized by high-resolution transmission electron microscopy, X-ray diffraction, magnetometry, and zeta potential measurements. The prepared nanomaterials were used for magnetic harvesting of microalgae. The highest harvesting efficiencies were found for PEI-coated Fe3O4. The efficiency was pH-dependent. Higher harvesting efficiencies, up to 99%, were obtained in acidic solutions. The results show that magnetic harvesting can be significantly enhanced by PEI coating, as it increases the positive electrical charge of the nanoparticles. Most importantly, the flocculants can be prepared at room temperature, thereby reducing the production costs.