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Algal biofuels are a promising alternative to fossil fuels, but their widespread use is hindered by problems with mass production. Light-emitting diodes (LEDs) with specific light wavelengths could be used as an energy source for algal growth and lipid synthesis. In this study, the effects of light source on the biomass and lipid production of the aerial microalga Coccomyxa sp. KGU-D001 were evaluated using LEDs. The integration of two-phase cultures, including growth and lipid production under the stress of nitrate depletion, was assessed for efficient lipid production under liquid- or aerial-phase conditions. Different wavelengths of light (blue, green, and red) were tested under liquid- and aerial-phase conditions. Under aerial-phase culture, the fatty acid contents in biofilm reached 320 mg g DWC -1 with the red LEDs. In view of these findings, we describe a one-step culture method for growth and lipid accumulation in algal biofilm under aerial-phase culture with red LED irradiation. When Coccomyxa biofilm was cultured on wet cotton wool with BBM in a petri dish under the red LED, it was able to grow and accumulate lipids under the aerial-phase condition. Based on the results of this study, a potential method for a continuous biodiesel production system is proposed., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The presence, biosynthesis and functional role of sterols in the green microalga Haematococcus pluvialis remain poorly understood. In this work we studied the effect of high-light (HL) stress on sterol synthesis in H. pluvialis UTEX 2505 cells. HL stress induced the synthesis of sterols in parallel with that of triacylglycerides (TAG), giving rise to the synthesis of cholesterol over that of phytosterols. Blockage of the carotenogenic 1-deoxy-D-xylulose 5-phosphate (MEP) pathway is shown to be involved in HL-induced sterol synthesis. In addition, high irradiance exposure induced MEP- and fatty acid (FA)-biosynthetic transcripts. The pharmacological inhibition of these pathways suggests a possible feedback regulation of sterol and FA homeostasis. Finally, both lipid classes proved crucial to the adequate photosynthetic performance of H. pluvialis grown under HL intensity stress. Our findings reveal new insights into H. pluvialis lipid metabolism that contribute to the development of value-added bioproducts from microalgae., Competing Interests: Declaration of competing interest All authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Microalgae are an alternative source for renewable energy to overcome the energy crises caused by exhaustion of fuel reserves. Algal biofuel technology demands a cost effective strategy for net profitable productivity. Inconsistent illumination intensities hinder microalgal growth. The light-utilizing efficiency of the cells is critical. Light scarcity leads to low production and high intensities cause photo-inhibition. We report effective usage of LEDs of different band wavelengths on the growth of microalgae in a closed, controlled environment to generate biomass and lipid yields. Among the different intensity and wavelengths tested. The light intensities of 500lx of blue-red combination gave maximum biomass in terms of cell density. LED of red light 220lx wavelength doubled the lipid dry weight from 30% (w/w) in white light to 60% (w/w). Thin layer lipid chromatogram demonstrated a dense and prominent spot of triacylglycerols in the red light, 220lx grown cultures. The FTIR profile indicates that different wavelength exposure did not alter the functional groups or change the chemical composition of the extracted lipids ensuring the quality of the product. We reiterate the fact that combination of red and blue LEDs is favoured over white light illumination for generation of biomass. In addition, we report an exciting finding of exposure to LEDs of red wavelength post-biomass generation lead to enhanced lipid production. This simple process doubled the lipid content harvested in 20days culture period., (Copyright © 2017 Elsevier B.V. All rights reserved.)

We attempted to enhance the growth and total lipid production of three microalgal species, Isochrysis galbana LB987, Nannochloropsis oculata CCAP849/1, and Dunaliella salina, which are capable of accumulating high content of lipid in cells. Low nitrogen concentration under photoautotrophic conditions stimulated total lipid production, but a decreasing total lipid content and an increasing biomass were observed with increasing nitrogen concentration. Among the different carbon sources tested for heterotrophic cultivation, glucose improved the growth of all three strains. The optimal glucose concentration for growth of I. galbana LB987 and N. oculata CCAP849/1 was 0.02 M, and that of D. salina was 0.05 M. Enhanced growth occurred when they were cultivated under heterotrophic or mixotrophic conditions compared with photoautotrophic conditions. Meanwhile, high total lipid accumulation in cells occurred when they were cultivated under photoautotrophic or mixotrophic conditions. During mixotrophic cultivation, biomass production was not affected significantly by light intensity; however, both chlorophyll concentration and total lipid content increased dramatically with increasing light intensity up to 150 µmol/m(2)/s. The amount and composition ratio of saturated and unsaturated fatty acids in cells were different from each other depending on both species and light intensity. The highest accumulation of total fatty acid (C16-C18) among the three strains was found from cells of N. oculata CCAP849/1, which indicates that this species can be used as a source for production of biodiesel.

In order to produce biodiesel from microalgae cultured with abundant seawater, Chlorella sp. was mutated with (137)Se-γ ray irradiation and domesticated with f/2 seawater culture medium (salinity=3 wt.%) under 15 vol.% CO2 stress. Biomass yield of the mutant increased by 25% compared with wild species and lipid content increased to 54.9%. When nitrogen and phosphorus concentrations in the initial substrate increased, the increased propagation speed of the mutant resulted in decreased cell diameter by 26.6% and decreased cell wall thickness by 69.7%. The dramatically increased biomass yield of the mutant with sufficient initial substrate and relative nitrogen starvation in the later growth period with continuous 15 vol.% CO2 led to an increased lipid yield of 1.0 g/L. The long-chain unsaturated fatty acids increased, whereas short-chain saturated fatty acids decreased., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Algal cultivation at high latitudes is challenged by the relatively low annual solar flux. One possible scenario to overcome this limitation is the use of mixotrophic growth to potentially boost biomass and lipid production. Here the effect of glycerol addition on the growth and lipid production by twelve indigenous microalgae was examined. The results show that there is considerable strain dependent variation in the maximum growth rate under mixotrophic conditions with the addition of glycerol causing in some cases up to a 2.4-fold increase in growth rate and a up to a 1.9-fold increase in biomass. In addition, glycerol increased total lipid production 40-60% in some strains. These results also show the value in screening culture collections for desired traits independent of strain identification since here one (PCH02) of the five Chlorella strains showed a large increase in lipid with glycerol., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The polarities of the cytoplasm and reactants were measured through dielectric spectroscopy, contact angle test, NMR, and FTIR to investigate the mechanisms underlying acid-catalyzed lipid transesterification in wet microalgal cells subjected to microwave irradiation. Organics with apolar functional groups in the cytoplasm decreased the contact angle of methanol against triglyceride by 13.92°, which subsequently increased transesterification efficiency by 2.4 times. The microalgal biomass, given its higher hydrophilicity index of 1.96 than lipids, was more accessible to hydrophilic alcohols, which subsequently promoted transesterification. Water in the cytoplasm promoted the dielectric constant of methanol and increased the contact angle of methanol against triglyceride by 20.51°, which subsequently decreased transesterification efficiency by 72.6%. The inhibitory effect of water on transesterification weakened with the prolonged carbon lengths of the alcohols because of decreased polarity. Microwave decreased the electric constants of alcohols and reduced the polarity difference between alcohols and lipids, thereby improving transesterification efficiency., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In this study, UV light was adopted to make it possible to attain sufficiently high extraction efficiency even with a minimal amount of H2O2. The Fenton-like reaction showed 80% of lipid extraction efficiency with 0.5% H2O2, whereas the provision of 16 W UV increased efficiency to 85% and decreased H2O2 consumption to 0.3%. This oxidation-based lipid extraction means have one fortuitous yet beneficial effect to remove chlorophylls, which are known to degrade the quality of the final product like biodiesel. The UV-Fenton-like reaction was found to eliminate 77% of chlorophylls. Such oxidation-based lipid extraction approaches as the Fenton-like reaction appear to have the sure application potential; and it is more so with the help of UV., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

NaCl is an effective inducer of lipid accumulation in freshwater microalgae, but little is known on whether the enhanced lipid components are desired. To address this issue, Monoraphidium dybowskii LB50 from a freshwater habitat was selected, cultivated, and subjected to NaCl induction at different scales outdoors. Results showed that the optimal salt concentration reduced glycolipid (GL) content, as well as enhanced neutral lipid (NL) and phospholipid (PL) contents. Moreover, GL was preferentially converted to NL at 20gL(-1) NaCl. Total lipid and NL contents respectively increased to 41.7% and 17.48% in 1d. The highest NL productivity was also achieved at both the 5L (24.13mgL(-1)d(-1)) and 140L (13.05mgL(-1)d(-1), 3.43gm(-2)d(-1)) scales. These results suggest that NL accumulated effectively and rapidly at different scales, indicating that this strategy has broad application prospects for the scale-up cultivation of oily algae., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The effects of ultrasonic treatment on fractal microstructures of wet microalgal cells were investigated for lipid extraction. Fractal dimension of cells with distorted surfaces increased with power and ultrasonication time. Microalgal cells shrank owing to dehydration and cytomembranes were reduced to debris, but cell walls were not fragmented. When ultrasonication power increased from 0 to 500W for 30min, the fractal dimension of cells increased from 1.21 to 1.51, cell sizes decreased from 2.78 to 1.68μm and cell wall thickness decreased from 0.08 to 0.05μm. When ultrasonication time increased from 5 to 30min with a power of 150W, the fractal dimension of cells increased from 1.24 to 1.37, cell sizes decreased from 2.72 to 2.38μm and cell wall thickness first increased to a peak of 0.22μm and then decreased. Long-chain and unsaturated lipids were degraded into short-chain and saturated lipids with ultrasonic cavitation., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Wavelength of light is a crucial factor which renders microalgae as the potential biodiesel. In this study, Tetraselmis sp. and Nannochloropsis sp. as famous targets were selected. The effect of different light wavelengths on growth rate and lipid production was studied. Microalgae were cultivated for 14 days as under blue, red, red-blue LED and white fluorescent light. The growth rate of microalgae was analyzed by spectrophotometer and cell counting while oil production under improved Nile red method. Optical density result showed the microalgae exhibited better growth curve under blue wavelength. Besides, Tetraselmis sp. and Nannochloropsis sp. under blue wavelength showed the higher growth rate (1.47 and 1.64 day(-1)) and oil production (102.954 and 702.366 a.u.). Gas chromatography analysis also showed that palmitic acid and stearic acid which were compulsory components for biodiesel contribute around 49-51% of total FAME from Nannochloropsis sp. and 81-83% of total FAME from Tetraselmis sp., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Microalgae are being investigated as potential candidates for biodiesel production since they can be grown without competition with food production, have an inherently fast growth rate, and can have a high lipid content under different nutrient limiting conditions. However, large scale production will best be carried out with indigenous strains, well adapted to local conditions. This study reports on the characterization of the novel microalga Chlorella sp. PCH90, isolated in Quebec. Its molecular phylogeny was established and lipid production studies as a function of the initial concentrations of nitrate, phosphate, and sodium chloride were carried out using response surface methodology. Under the appropriate conditions this microalga could produce up to 36% lipid and grew well in both synthetic medium and secondary effluent from a wastewater treatment plant at both 22 and 10°C. Thus, this strain is promising for further development as a potential biofuels producer under local climatic conditions., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The optimal conditions for cultivating the thermotolerant lipid-rich microalga Desmodesmus sp. F2 to achieve maximal lipid productivity were determined in this study. The conditions were light intensity, 700μmol/m(2)s; temperature, 35°C; cultivation nitrogen source, nitrate; initial nitrogen level, 6.6mM nitrogen. Carbon dioxide (2.5%, 0.2 vvm) was pumped into the cultures continuously. In the pre-optimized conditions, the maximal lipid productivity of this microalga was 113mg/L/d, which was raised to 263mg/L/d in the optimized conditions. This level of lipid productivity of microalgae is the highest ever reported in the literature. Fatty acid composition of the lipid produced by Desmodesmus sp. F2 in the optimal conditions was determined, in which C16 and C18 species accounted for 95% of the fatty acids. Saturated, monounsaturated and polyunsaturated fatty acids accounted for 38.9%, 33.1% and 22.6%, respectively. Based on the analysis, this lipid quality makes it a good feedstock for biodiesel production., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

This study explores the influence of alkaline pH and light intensity on the performance of Neochloris oleoabundans in two-stage batch cultivation: a first stage for nitrogen-sufficient growth followed by a second stage for lipid accumulation under nitrogen starvation. The highest TAG yield on absorbed light was obtained at low light conditions when pre-cultivation occurred at pH 8 and lipid accumulation was induced at pH 10. However, a higher alkaline pH by itself appears not to enhance the starvation-induced increase in lipid contents, except when combined with high light and pre-cultivation occurs at those same conditions. Such strategy however also results in low biomass and TAG yields on absorbed light. Fatty acid composition analysis revealed that the relative fatty acid contents of the TAG pool are nevertheless independent from the light intensity and pH applied at either cultivation stage, suggesting a high specificity of N. oleoabundans cell machinery towards TAG production., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Light quality and the intensity are key factors which render microalgae as a potential source of biodiesel. In this study the effects of various intensities of blue light and its photoperiods on the growth and lipid content of Chlorella vulgaris were investigated by using LED (Light Emitting Diode) in batch culture. C. vulgaris was grown for 13 days at three different light intensities (100, 200 and 300 μmol m(-2)s(-1)). Effect of three different light and dark regimes (12:12, 16:08 and 24:00 h Light:Dark) were investigated for each light intensity at 25°C culture temperature. Maximum lipid content (23.5%) was obtained due to high efficiency and deep penetration of 200 μmol m(-2)s(-1) of blue light (12:12 L:D) with improved specific growth (1.26 d(-1)) within reduced cultivation time of 8 days. White light could produce 20.9% lipid content in 10 days at 16:08 h L:D., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Nannochloropsis has been considered as a promising feedstock for biodiesel production in recent years. To improve its lipid productivity, heavy-ion irradiation mutagenesis, an effectively breeding method used in plants and microorganisms was applied in Nannochloropsis oceanica OZ-1. After large-scale screening using Imaging-PAM and microplate-reader, two mutants (HP-1 and HP-2) with higher growth rate were isolated from the wild type N. oceanica. Subsequently analysis showed that after 18 days of cultivation biomass accumulation of the HP-1 and HP-2 mutant was increased by 18% and 26% respectively compare to the wild type. Total lipid productivity of the HP-1 and HP-2 mutant was 295 mg/(L x d) and 275 mg/(L x d), respectively, whereas that of the wild type was 247 mg/(L x d). Both mutants showed significantly advantage over their wild type concerning biomass accumulation and lipid productivity.

Microalgae are a promising feedstock for biofuels because of their capability to produce lipids. Cell disruption is necessary to maximize lipid extraction. Sonication conditions were evaluated for breaking heterotrophic (Schizochytrium limacinum) and autotrophic (Chlamydomonas reinhardtii) microalgae cells. Cell disruption was estimated by Nile red-lipids fluorescence quantification in S. limacinum and by the release of intracellular chlorophyll and carotenoids in green microalga C. reinhardtii. In both species, approximately 800 J/10 mL was the energy input necessary to maximize cell disruption, regardless of the cell concentrations studied. Increasing sonication time produced increasing amount of free radicals, quantified by the formation of hydroxyterephthalate. Sonication energy beyond the level needed for cell disruption induced oxidation of arachidonic acid, a polyunsaturated fatty acid typically found in marine lipids. Careful control of sonication conditions is necessary to maximize oil extraction at the lowest operational cost and to prevent oil from free radical-induced degradation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

For overcoming the long period of seed cultured photoautotrophically and inadequate cell supply for the inoculation of microalgae photoautotrophic cultivation, a model for the photoautotrophic culture of three Chlorella species with heterotrophic cells as seed was investigated. The model can not only take advantages of rapid cell growth in heterotrophic process for preparation of cells as seed but also increase the biomass and lipid productivities of the microalgae cultivated photoautotrophically. The results showed that biomass productivities of Chlorella pyrenoidosa, Chlorella ellipsoidea and Chlorella vulgaris cultured by heterotrophy were 20.9, 26.9 and 25.2 times higher than those by photoautotrophy in seed culturing period. In the subsequent photoautotrophic culture, the biomass and lipid productivities of C. pyrenoidosa, C. ellipsoidea and C. vulgaris with heterotrophic seed were 1.91, 1.51, 1.48 and 1.66, 1.37, 1.42 times higher than those with photoautotrophic seed. Furthermore, the culture model was also carried out successfully outdoor., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The green microalga strain, Botryococcus braunii KMITL 2, was isolated from a freshwater reservoir in central Thailand, and the effects of light, nitrogen, phosphorus, iron, cultivation time and salinity on lipid production were studied by varying parameters one at a time. When cultured in Chlorella medium containing 222 mg L(-1) phosphorus (PO(4)(3-)-P) under continuous illumination of 200 μE m(-2)s(-1) with a salinity of 0 psu, a maximum lipid content of 54.69 ± 3.13% was obtained. Its high lipid content makes strain KMITL 2 a potential source for biodiesel production in tropical regions., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

We examined responses of batch cultures of the marine microalga Nannochloropsis sp. to combined alterations in salinity (13, 27, and 40 g/l NaCl) and light intensity (170 and 700 μmol photons/m(2)·s). Major growth parameters and lipid productivity (based on total fatty acid determination) were determined in nitrogen-replete and nitrogen-depleted cultures of an initial biomass of 0.8 and 1.4 g/l, respectively. On the nitrogen-replete medium, increases in light intensity and salinity increased the cellular content of dry weight and lipids due to enhanced formation of triacylglycerols (TAG). Maximum average productivity of ca. 410 mg TFA/l/d were obtained at 700 μmol photons/m(2)·s and 40 g/l NaCl within 7 days. Under stressful conditions, content of the major LC-PUFA, eicosapentaenoic acid (EPA), was significantly reduced while TAG reached 25% of biomass. In contrast, lower salinity tended to improve major growth parameters, consistent with less variation in EPA contents. Combined higher salinity and light intensity was detrimental to lipid productivity under nitrogen starvation; biomass TFA content, and lipid productivity amounted for only 33% of DW and ca. 200 mg TFA/l/day, respectively. The highest biomass TFA content (ca. 47% DW) and average lipid productivity of ca. 360 mg TFA/l/day were achieved at 13 g/l NaCl and 700 μmol photons/m(2)·s. Our data further support selecting Nannochloropsis as promising microalgae for biodiesel production. Moreover, appropriate cultivation regimes may render Nannochloropsis microalgae to produce simultaneously major valuable components, EPA, and TAG, while sustaining relatively high biomass growth rates.