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To facilitate the lipid extraction from Nannochloropsis oceanica with thick cell wall using switchable hydrophilicity solvent, ultrasound-assisted N, N, N', N'-tetraethyl-1,3-propanediamine (TEPDA) was used to effectively destruct the cell wall. TEPDA cations were adsorbed on the cells via electrostatic force and formed the electron-donor-acceptor (EDA) complex with the hydroxyl groups in cellulose. This broke the hydrogen-bonding interactions between cellulose chains and stripped them from cell wall, thus reducing the cell wall thickness from 141 nm to 68.6 nm. Moreover, TEPDA cations neutralized the negatively charged phospholipid bilayers, decreasing the cell surface zeta potential from -27.5 eV to -14.1 eV. The local electrostatic equilibrium led to cell membrane leakage. The ultrasound promoted the stripping of the cellulose chains at a power intensity of 0.5 W/mL and frequency of 20 kHz, achieving the lipid extraction efficiency of 98.2% within 2 h at a volume ratio of 1:4 of wet microalgae to TEPDA., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

In order to reutilize Fe 2 O 3 particles in flue gas from coal-fired power plant as a ferrum nutrient for improving microalgae growth, Na-Citrate was proposed to chelate FeCl 3 derived from Fe 2 O 3 and HCl reactions to promote biomass and lipid productivities of Chlorella PY-ZU1. Fe-Citrate gave much higher biomass and lipid productivities than FeCl 3 , Fe-EDTA, Fe-DTPA and Fe-HEDTA, because organic chelator prevented Fe 3+ from depositing, lower stability constant resulted in easier dissociation of ferric chelate, smaller chelate facilitated Fe 2+ (reduced from Fe 3+ ) transportation through cell membranes. The biomass growth and photosynthetic capacity of Chlorella PY-ZU1 cultivated with Fe-Citrate (converted from Fe 2 O 3 particles) medium were similar to those with commercial ferric ammonium citrate medium. The biomass and lipid productivities of Chlorella PY-ZU1 cultivated with 5 mg L -1 Fe-Citrate medium were 1.30 and 1.72 times, respectively, higher than those with FeCl 3 growth medium., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Computational fluid dynamics were used to analyze the flash light effect and CO 2 bubble behavior in an optimized flow field generated by double paddlewheels in a flat plate photoreactor to enhance microalgal biomass productivity. The increased D/w ratio significantly enhanced the average turbulent kinetic energy and flash cycle frequency. However, the effects became weak when the D/w ratio was over 0.67. Appliance of double paddlewheels increased flash cycle frequency from 0.035 to 0.121 Hz and increased light time ratio from 8.3% to 31.5%. Meanwhile, the bubble dynamic behavior was characterized using population balance model. The average bubble size reduced by 24.4% and the bubble rising velocity reduced by 10.6%, which facilitated CO 2 mixing and mass transfer in microalgal solution. Therefore, biomass accumulation of microalgae Chlorella in the photoreactor with double paddlewheels increased by 62.3% under 15% CO 2 ., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Switchable solvent N, N, N', N'-tetraethyl-1,3-propanediamine (TEPDA) was proposed to extract lipids from wet Nannochloropsis oceanica with a 5% higher extraction efficiency than chloroform-methanol. It was found that TEPDA acted mainly as an organic solvent to soften and dissolve lipids, while a small amount of TEPDA was dissociated into tertiary amine ion, i.e.,(C 2 H 5 ) 2 N-(CH 2 ) 3 -NH + (C 2 H 5 ) 2 . This cation acted as a surfactant to promote cell disruption and lipid separation. With moisture increasing from 0 to 84 wt%, more TEPDA was dissociated into cationic surfactant to induce local rearrangement of phospholipid bilayers in cell membranes through electrostatic interaction, resulting in the fractal dimension of disrupted cells increased from 1.49 to 1.66. Accordingly, the yield of fatty acid methyl ester (FAME) through transesterification of lipids extracted with TEPDA increased by 9%, while FAME yield from lipids extracted with chloroform and n-hexane decreased by 41% and 65%, respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Chemical flocculation is considered to be an overall low-cost and up-scalable process for harvesting of microalgae. In this study a new flocculation approach utilizing metal coagulant (Fe 2 (SO 4 ) 3 ) and sulfuric acid (H 2 SO 4 ) was introduced for harvesting of Chlorella sp. KR-1, which overcome two main issues of contamination and reuse of coagulant. Reduction of pH successfully released precipitates attached to the microalgae, and the remaining acidic solution containing recovered ferric ions could be reused for harvesting up to three times with high, better-than 98% efficiencies. Moreover, the acid-treated microalgal biomass could be directly used for lipid extraction without additional catalyst. High extraction yields of around 32% were achieved with FAME conversion efficiencies of around 90%. The integrated approach devised in the present study is expected to make the best use of the age-old yet effective harvesting means of flocculation, which can be a practical and economical option in microalgal biorefinery., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Recently, biofuels and nutraceuticals produced from microalgae have emerged as major interests, resulting in intensive research of the microalgal biorefinery process. In this paper, recent developments in cell-wall disruption and extraction methods are reviewed, focusing on lipid and astaxanthin production from the biotechnologically important microalgae Chlorella and Haematococcus, respectively. As a common, critical bottleneck for recovery of intracellular components such as lipid and astaxanthin from these microalgae, the composition and structure of rigid, thick cell-walls were analyzed. Various chemical, physical, physico-chemical, and biological methods applied for cell-wall breakage and lipid/astaxanthin extraction from Chlorella and Haematococcus are discussed in detail and compared based on efficiency, energy consumption, type and dosage of solvent, biomass concentration and status (wet/dried), toxicity, scalability, and synergistic combinations. This report could serve as a useful guide to the implementation of practical downstream processes for recovery of valuable products from microalgae including Chlorella and Haematococcus., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In this study, a simultaneous process of harvesting biomass and extracting crude bio-oil was attempted from wet microalgae biomass using FeCl3 and Fe2(SO4)3 as both coagulant and cell-disrupting agent. A culture solution of Chlorella sp. KR-1 was firstly concentrated to 20 g/L and then proceeded for cell disruption with the addition of H2O2. Optimal dosage were 560 and 1060 mg/L for FeCl3 and Fe2(SO4)3, showing harvesting efficiencies of more than 99%. Optimal extraction conditions were identified via the response surface method (RSM), and the extraction yield was almost the same at 120 °C for both iron salts but FAME compositions after transesterification was found to be quite different. Given iron salts were a reference coagulant in water treatment in general and microalgae harvesting in particular, the present approach of using it for harvesting and oil-extraction in a simultaneous manner can serve as a practical route for the microalgae-derived biodiesel production., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In this study, hydrothermal acid treatment for efficient recovery of sugar from Golenkinia sp. was investigated. The initial glucose and XMG (xylose, mannose, and galactose) contents of a prepared Golenkinia sp. solution (40g/L) were 15.05 and 5.24g/L, respectively. The microalgal cell walls were hydrolyzed, for sugar recovery, by enzymatic saccharification and/or hydrothermal acid treatment. Among the various hydrothermal acid treatment conditions, the most optimal were the 2.0% H2SO4 concentration at 150°C for 15min, under which the glucose- and XMG-extraction yields were 71.7% and 64.9%, respectively. By pH 4.8, 50°C enzymatic hydrolysis after optimal hydrothermal acid treatment, the glucose- and XMG-extraction yields were additionally increased by 8.3% and 0.8%, respectively. After hydrothermal acid treatment, the combination with the enzymatic hydrolysis process improved the total sugar yield of Golenkinia sp. to 75.4%., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

An interest in biodiesel as an alternative fuel for diesel engines has been increasing because of the issue of petroleum depletion and environmental concerns related to massive carbon dioxide emissions. Researchers are strongly driven to pursue the next generation of vegetable oil-based biodiesel. Oleaginous microalgae are considered to be a promising alternative oil source. To commercialize microalgal biodiesel, cost reductions in oil extraction and downstream biodiesel conversion are stressed. Herein, starting from an investigation of oil extraction from wet microalgae, a review is conducted of transesterification using enzymes, homogeneous and heterogeneous catalysts, and yield enhancement by ultrasound, microwave, and supercritical process. In particular, there is a focus on direct transesterification as a simple and energy efficient process that omits a separate oil extraction step and utilizes wet microalgal biomass; however, it is still necessary to consider issues such as the purification of microalgal oils and upgrading of biodiesel properties., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Hydrothermal acid (combined with autoclaving and nitric acid) pretreatment was applied to Nannochloropsis salina as a cost-effective yet efficient way of lipid extraction from wet biomass. The optimal conditions for this pretreatment were determined using a statistical approach, and the roles of nitric acid were also determined. The maximum lipid yield (predicted: 24.6%; experimental: 24.4%) was obtained using 0.57% nitric acid at 120°C for 30min through response surface methodology. A relatively lower lipid yield (18.4%) was obtained using 2% nitric acid; however, chlorophyll and unsaturated fatty acids, both of which adversely affect the refinery and oxidative stability of biodiesel, were found to be not co-extracted. Considering its comparable extractability even from wet biomass and ability to reduce chlorophyll and unsaturated fatty acids, the hydrothermal nitric acid pretreatment can serve as one direct and promising route of extracting microalgae oil., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Microalgal lipid with a high free fatty acid (FFA) content was directly extracted from Chlorella vulgaris, using SDBS, in an acid-catalyzed hot-water extraction process. The total fatty acid content of C. vulgaris was 296.0 mg/g cell. Under the 1.0% sulfuric acid, 0.4% SDBS conditions, the FFA content of the lipid increased to 96.7%, and the lipid-extraction yield was 248.4 mg/g cell. Under the 2.0% sulfuric acid, 0.2% SDBS conditions, the FFA content of the lipid was 96.1%, and the lipid-extraction yield was 266.0mg/g cell. Whereas the FAME content of the microalgal lipid extracted by hexane-methanol was 76.4% at the 10.0% sulfuric acid concentration, the FAME content of the high-FFA microalgal lipid was increased to 70.1% at a sulfuric acid concentration of only 0.1%. By combined sulfuric acid/SDBS treatment, high-FFA microalgal lipid was extracted in large yields; moreover, the amount of catalyst was remarkably reduced in the esterification of FFA., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, acid-catalyzed hot-water extraction of docosahexaenoic acid (DHA)-rich lipids from Aurantiochytrium sp. was performed, and its yield-enhancing effects were investigated. The total fatty acid content of the Aurantiochytrium sp. was 482.5mg/g cell, of which 141.7mg/g cell (29.4% of total fatty acids) was DHA. The lipid-extraction yield by acid-catalyzed hot-water treatment was compared with those by organic solvents. Among the various acid-catalyzed hot-water treatment conditions, the most optimal were 1.00% H2SO4 concentration, 100°C, 30min, under which the lipid-extraction yield was 472.4mg/g cell, and most of the DHA was extracted (29.2% of total fatty acids). Acid-catalyzed hot-water extraction treatment markedly improved the lipid-extraction yield of Aurantiochytrium sp., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Microalgae biorefinement has attracted in intensive academic and industrial interest worldwide for its potential to replace petrol biofuels as economically and environmentally advantageous alternatives. However, harvesting and lipid extraction remain as critical and difficult issues to be resolved. In the present study, four amino-groups functionalized organic-nano clays were prepared. Specifically, Mg or Al or Ca backboned and covalently linked with 3-aminopropyltriethoxysilane or 3-[2-(2-aminoethylamino)ethylamino]propyltrimethoxysilane by sol-gel reaction under ambient conditions, resulted in Mg-APTES clay, Al-APTES clay, Ca-APTES clay, and Mg-N3 clay, respectively. Each organic-nanoclay was utilized for lipid extraction from wet microalgae biomass. As a result, the lipid-extraction efficiency of paste docosahexaenoic acid (DHA)-rich Chlorella sp. with low lipid content was high, while one of paste oleaginous Chlorella sp. with high lipid content was relatively low. Despite the low lipid-extraction efficiencies in all of the wet microalgae biomass, the conversion of the extracted lipids' fatty acids to biodiesel was nearly 100%., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Microalgal cultivation using wastewater is now regarded as essential for biodiesel production, as two goals can be achieved simultaneously; that is, nutrient removal efficiency and biomass production. Therefore, this study examined the effects of carbon sources, the N:P ratio, and the hydraulic retention time (HRT) to identify the optimal conditions for nutrient removal efficiency and biomass production. The effluent from a 2nd lagoon was used to cultivate microalgae. Whereas the algal species diversity and lipid content increased with a longer HRT, the algal biomass productivity decreased. Different carbon sources also affected the algal species composition. Diatoms were dominant with an increased pH when bicarbonate was supplied. However, 2% CO(2) gas led to a lower pH and the dominance of filamentous green algae with a much lower biomass productivity. Among the experiments, the highest chlorophyll-a concentration and lipid productivity were obtained with the addition of phosphate up to 0.5 mg/l P, since phosphorus was in short supply compared with nitrogen. The N and P removal efficiencies were also higher with a balanced N:P ratio, based on the addition of phosphate. Thus, optimizing the N:P ratio for the dominant algae could be critical in attaining higher algal growth, lipid productivity, and nutrient removal efficiency.

Highlights: [•] Chlorophyll removal from microalgal lipid by hypochlorous acid. [•] Decrease in unsaturated fatty acid content of microalgal lipid by hypochlorous acid. [•] Hypochlorous acid produced from NaClO solution and NaCl electrolysis. [Copyright &y& Elsevier]

Graphical abstract Highlights • Comparison of imidazolium ionic liquids (ILs) for disruption of H. pluvialis cyst. • Co-extraction of lipid and astaxanthin using room-temperature IL/water mixtures. • Very high astaxanthin recoveries (>99%) by IL pretreatments and hexane extractions. Abstract Energy-saving, high-efficiency cell disruption is a critical step for recovery of thermolabile antioxidant astaxanthin from Haematococcus pluvialis cyst cells of rigid cell-wall structure. In this study, as room-temperature green solvents, 10 types of 1-ethyl-3-methylimidazolium ([Emim])-based ionic liquids (ILs) were compared and evaluated for their abilities to disrupt H. pluvialis cyst cells for astaxanthin/lipid extraction. Among the 10 ILs tested, 3 [Emim]-based ILs with HSO 4 , CH 3 SO 3 , and (CF 3 SO 2) 2 N anions were selected based on astaxanthin/lipid extraction performance and synthesis cost. When pretreated with IL/water mixtures, intact cyst cells were significantly torn, broken or shown to release cytoplasmic components, thereby facilitating subsequent separation of astaxanthin/lipid by hexane. However, excess IL pretreatments at high temperature/IL dosages and longer incubation times significantly deteriorated lipid and/or astaxanthin. Under optimized mild conditions (6.7% (v/v) IL in water solution, 30 °C, 60 min), almost complete astaxanthin recoveries (>99%) along with moderate lipid extractions (∼82%) could be obtained. [ABSTRACT FROM AUTHOR]

• Novel bifunctional heterogeneous catalyst was synthesized for biodiesel production. • IL/IRMOF-3 exhibited synergistic catalysis of acid-base and Brønsted-Lewis acid sites. • The new bond formed between IL and IRMOF-3 strengthened catalysts recyclability. • The conversion efficiency of microalgal lipids to FAME with IL/IRMOF-3 was 98.2% In order to enhance catalytic active sites to improve microalgal biodiesel production, a novel efficient bifunctional catalyst was synthesized by modifying metal–organic framework (IRMOF-3) with Brønsted acidic ionic liquid (1-butylsulfonate-3-methylimidazolium bisulfate [BSO 3 HMIm][HSO 4 ], IL). It was determined that IL was immobilized on IRMOF-3 framework through chemical bonding formed by an acid-base reaction between sulfonic acid group of IL and amino group of IRMOF-3. The bifunctional catalyst exhibited acid–base synergistic catalysis as well as a synergistic effect of Brønsted acid and Lewis acid, which efficiently catalyzed esterification and transesterification of microalgal lipids. The introduction of [BSO 3 HMIm][HSO 4 ] on IRMOF-3 framework promoted the acidity of the catalyst from 8.91 to 10.76 mmol/g and increased strong/weak acidity ratio from 0.27 to 0.85, resulting an increased catalytic conversion efficiency of 98.2% under optimal reaction conditions. Due to the strong ionic bond, the catalyst showed good reusability with the catalytic efficiency up to 85.5% after six cycles. [ABSTRACT FROM AUTHOR]

In the present study, a simple magnetic-particle recycling strategy was developed for harvest of the oleaginous microalga Chlorella sp. KR-1. The method entails the flocculation of microalgal cells and bare-Fe3O4 magnetic particles (bMP) by electrostatic attraction and the subsequent recovery of the bMP from the harvested flocs by electrostatic repulsion below and above the isoelectric points (IEP), respectively. For 10 recycles, the bMP showed 94-99% and 90-97% harvest and recovery efficiencies, respectively. Furthermore, neither the use of bMP nor pH adjustment showed any adverse effect on the microalgal cell growth or the co-existing bacterial species, as confirmed from the subsequent medium-recycling test and denaturing gradient gel electrophoresis (DGGE) analysis. [ABSTRACT FROM AUTHOR]

Abstract: In this study, pre-biodiesel-production lipid extraction from microalgae (Chlorella vulgaris) was performed. The yield-enhancing effects of ionic liquid blends on the lipid extraction were investigated. The initial fatty acids content of the C. vulgaris was 292.2 mg/g cell. The lipid extraction yield using single ionic liquids was compared with the yield obtained with organic solvents and ionic liquid mixtures, respectively. The yield by hexane–methanol solvent was 185.4 mg/g cell. Among the 12 ionic liquids, 1-ethyl-3-methyl imidazolium acetate, 1-ethyl-3-methyl imidazolium diethylphosphate, 1-ethyl-3-methyl imidazolium tetrafluoroborate, and 1-ethyl-3-methyl imidazolium chloride showed high (>200.0 mg/g cell) lipid extraction yields. Although the yields of 1-ethyl-3-methyl imidazolium ethyl sulfate and 1-ethyl-3-methyl imidazolium thiocyanate were only 60.5 and 42.7 mg/g cell, respectively, the yield for their mixture (weight ratio of 1:1) was improved to 158.2 mg/g cell. Similarly, whereas the lipid extraction yield of 1-ethyl-3-methyl imidazolium hydrogen sulfate was just 35.2 mg/g cell, that for its mixture with 1-ethyl-3-methyl imidazolium thiocyanate (weight ratio of 1:1) was boosted to 200.6 mg/g cell. Overall, the synergistic effects of the ionic liquid mixtures with different anions improved the lipid extraction yield of C. vulgaris. [Copyright &y& Elsevier]

Highlights: [•] Aminoclay-induced humic acid flocculation (AC-HA) was investigated. [•] AC-HA showed for efficient harvesting of oleaginous microalgae. [•] AC-HA played a type of network in capturing microalgal cells. [•] Flocculation approach can be utilized for microalgae flocculation. [Copyright &y& Elsevier]

Abstract: In this study, the effects of enzymatic hydrolysis on lipid extraction from microalga (Chlorella vulgaris) were investigated prior to biodiesel production. The initial fatty acids content of C. vulgaris was 87.6 mg/g cell. The microalgal cell walls were hydrolyzed by cellulases and then their lipid fractions were extracted using various organic solvents such as hexane, methanol, and chloroform. Optimal pH and temperature for the enzymatic hydrolysis were pH 4.8 and 50 °C, respectively, and the maximal hydrolysis yield was 85.3%, which was achieved after 72 h. After the enzymatic hydrolysis, the lipid extraction yield by the organic solvents was improved compared to when there was no enzymatic hydrolysis process, by 1.29–1.73-fold depending on the solvents used. The total fatty acid methyl ester (FAME) productivity through the enzymatic hydrolysis was higher than when there was no enzymatic hydrolysis, by 1.10–1.69-fold depending on the solvents used. When lipid was extracted from the C. vulgaris after the enzymatic hydrolysis in chloroform-methanol solution, FAME productivity was 59.4 mg FAME/g cell. [Copyright &y& Elsevier]

Abstract: In microalgae-based biorefinement, one of the highest practical priorities is to reduce the costs of downstream processes. As one potential solution, microalgae harvesting by organoclays has received particularly keen research interest. In the present study, cationic charged aluminum- and magnesium-backboned organoclays were synthesized and solubilized in aqueous solution due to their high-density of amino sites. Each, within 30min of its injection into 1.7g/L-concentration microalgal feedstocks, effected harvesting efficiencies of almost 100% at concentrations above 0.6g/L while maintaining a neutral pH. Conclusively, organoclays, if recycled efficiently, can be uniquely effective microalgae harvesting agents. [Copyright &y& Elsevier]

• ZIF-8 nanoparticles promoted dissolving carbon in a photo-bioreactor. • ZIF-8 with small size was more effective at increasing mass transfer coefficient. • Biomass accumulation was increased by 25.5% with ZIF-8 nanoparticles. Low dissolution of CO 2 in closed tubular photobioreactors results in insufficient carbon source supply, and often limits microalgae growth. An optimized ZIF8-SE medium containing zeolitic imidazolate framework-8 (ZIF-8) nanoparticles was developed for strengthening CO 2 mass transfer in the photobioreactor, which improved CO 2 dissolution to increase microalgal biomass yield under 15% CO 2. The CO 2 mass transfer coefficient markedly increased from 1.82 to 3.85, leading to an increased dissolved inorganic carbon concentration. Meanwhile, microalgae biomass accumulation with 0.01 mmol/L ZIF-8 nanoparticles was 25.5% higher than that without ZIF-8 nanoparticles. Furthermore, the average cell size decreased from 2.61 to 2.38 μm and the cell fractal dimension increased from 1.38 to 1.56 when ZIF-8 nanoparticle concentration increased from 0 to 0.2 mmol/L, indicating that potential toxicity might occurred with excess ZIF-8 nanoparticles. Reutilization of ZIF-8 nanoparticles in the recycled culture medium improved microalgal growth by 23%, which was similar to the promotion of fresh ZIF8-SE medium. This new finding provides a new strategy to improve CO 2 mass transfer in the horizontal tubular photobioreactor, which makes it a feasible platform for CO 2 capture and utilization. [ABSTRACT FROM AUTHOR]