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High moisture content in wet algal biomass hinders effective performance of current lipid extraction methods. An improved aqueous extraction method combing thermal and enzymatic lysis was proposed and performed in algal slurry of Nannochloropsis oceanica (96.0% moisture) in this study. In general, cell-wall of N. oceanica was disrupted via thermal lysis and enzymatic lysis and lipid extraction was performed using aqueous surfactant solution. At the optimal conditions, high extraction efficiencies for both lipid (88.3%) and protein (62.4%) were obtained, which were significantly higher than those of traditional hexane extraction and other methods for wet algal biomass. Furthermore, an excessive extraction of polar lipid was found for wet biomass compared with dry biomass. The advantage of this method is to efficiently extract lipids from high moisture content algal biomass and avoid using organic solvent, indicating immense potential for commercial microalgae-based biofuel production. [ABSTRACT FROM AUTHOR]

A mixed culture of microalgae, containing two Scenedesmus species, was analysed to determine its potential in coupling of pulp and paper mill effluent treatment and microalgal cultivation. Laboratory studies suggested that 60% concentration of wastewater was optimum for microalgal cultivation. A maximum of 82% and 75% removal of BOD and COD respectively was achieved with microalgal cultivation in outdoor open pond. By the end of the cultivation period, 65% removal of NO 3 -N and 71.29% removal of PO 4 -P was observed. The fatty acid composition of mixed microalgal culture cultivated with effluent showed the palmitic acid, oleic acid, linoleic acid and α-linolenic acid as major fatty acids. The results obtained suggest that pulp and paper mill effluent could be used effectively for cultivation of microalgae to minimise the freshwater and nutrient requirements. [ABSTRACT FROM AUTHOR]

This study fundamentally investigated the pseudo-catalytic transesterification of dried Nannochloropsis oceanica into fatty acid methyl esters (FAMEs) without oil extraction, which was achieved in less than 5 min via a thermo-chemical pathway. This study presented that the pseudo-catalytic transesterification reaction was achieved in the presence of silica and that its main driving force was identified as temperature: pores in silica provided the numerous reaction space like a micro-reactor, where the heterogeneous reaction was developed. The introduced FAME derivatization showed an extraordinarily high tolerance of impurities ( i.e ., pyrolytic products and various extractives). This study also explored the thermal cracking of FAMEs derived from N. oceanica : the thermal cracking of saturated FAMEs was invulnerable at temperatures lower than 400 °C. Lastly, this study reported that N. oceanica contained 14.4 wt.% of dried N. oceanica and that the introduced methylation technique could be applicable to many research fields sharing the transesterification platform. [ABSTRACT FROM AUTHOR]

Conditions of light-emitting diode (LED) wavelength, light intensity, nitrate concentration, and time of exposure to green LED light stress in a two-phase culture were optimized for lipid production with three species of microalgae. Among the three microalgae, Nannochloropsis oculata showed the highest specific growth rate ( μ max ) of 0.037 h −1 and the lowest saturation constant ( K s ) of 1.32 mg/L. The highest lipid contents of the three microalgae in the second phase under green LED light stress were 52.0% (w/w) for Nannochloropsis salina at 2 days, 53.0% (w/w) for Nannochloropsis oceanica at 2 days, and 56.0% for N. oculata at 2 days. Fatty acid analysis of the microalgae showed that 85–87% (w/w) of total fatty acids from N. salina , N. oceanica , and N. oculata consisted of palmitic acid (C16:0) and oleic acid (C18:1). [ABSTRACT FROM AUTHOR]

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 [NaNO 3 , NH 4 HCO 3 , MgSO 4 ·7H 2 O, KH 2 PO 4 , K 2 HPO 4 and (NH 4 ) 2 HPO 4 ], 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 (0.73 g L −1 ) compared with the BBM medium and 85% the lipid concentration (281 mg L −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. [ABSTRACT FROM AUTHOR]

A statistical mixture design approach was used to investigate the effects of cell wall degrading enzymes on the recovery of lipids from Nannochloropsis sp. A preliminary screening of potentially suitable enzyme preparations, including lysozyme, cellulase and different types of hemicellulases, was carried out. The most effective preparations were then taken as basic components for the formulation of enzyme mixtures. Optimized ternary mixtures consisting of cellulase and two hemicellulases were obtained which allowed the recovery of up to 37.2 g of lipids per 100 g of dry biomass. SEM and TEM images of the enzymatically treated microalga revealed extensive cell damage, with degradation of the cell wall and release of intracellular material. Overall, the results obtained demonstrate that the mixture design method can be used to prepare cell wall degrading enzyme cocktails that can significantly improve the recovery of lipids or other valuable components from microalgae. [ABSTRACT FROM AUTHOR]

Taxonomy and phylogeny of twenty two microalgal isolates were examined using both universal and newly designed molecular primers. Among the isolates, Scenedesmus bijugus , Coelastrella sp., Auxenochlorella protothecoides , and Chlorella sp. were particularly promising in terms of producing lipids as measured by fatty acid methyl esters (FAME) analysis and significant concentration of carotenoids. A comparative experiment showed that S. bijugus and Chlorella sp. were the most promising candidates (L − 1 d − 1 , with biomass) 174.77 ± 6.75, 169.81 ± 5.22 mg, lipids 40.14 ± 3.31, 39.72 ± 3.89 mg, lutein 0.47, 0.36 mg, and astaxanthin 0.27, 0.18 mg respectively. The fatty acids produced by these microalgal isolates were mainly palmitic, stearic, oleic, linoleic, and linolenic acid. The freshwater microalgal isolate S. bijugus be the most suitable isolate for producing biodiesel and carotenoids, due to high productivity of biomass, lipids, metabolites, and its suitable fatty acid profile. [ABSTRACT FROM AUTHOR]

A two-step strategy comprising heterotrophic cultivation and photo-chemical modulator induction was developed to enhance biomass and lipid accumulation in the oleaginous Monoraphidium sp. QLY-1, which was isolated from Qilu Lake in Yunnan Plateau. The algae were first cultivated heterotrophically to achieve high biomass concentration (5.54 g L −1 ) with a lipid content of 22.47%. The cultivated algae were diluted, transferred to light environment, and treated with different chemical elicitors. Results showed that the lipid content increased to 36.68% after 3-day of photoinduction. The lipid content was further enhanced by 1.21, 1.32, and 1.29 folds in algal cells treated with nitrogen deficiency, 20 g L −1 NaCl, and 5 mM glycine betaine, respectively. The maximum lipid content (48.54%) and lipid productivity (121.27 mg L −1 d −1 ) were obtained in treatments with 20 g L −1 NaCl and 5 mM GB, respectively. This study proposes a strategy to efficiently produce lipids by using microalgae. [ABSTRACT FROM AUTHOR]

Despite the vast interest in microalgae as feedstock for biodiesel production, relatively few studies examined their response to diurnal temperature fluctuation. Here, we describe biomass and lipid productivities and fatty acid profiles of thermo-resistant Micractinium sp. and Scenedesmus sp. grown in batch cultures in a laboratory set-up that mimics a typically warm summer day in Central Anatolia with a 16-h light temperature of 30 °C and 8-h dark temperature of 16 °C (30 °C (day)/16 °C (night)). Both strains can survive a temperature range of 10–50 °C. We found the lipid productivities of Micractinium sp. and Scenedesmus sp. as 30/21 mg L −1 d −1 and 6/7 mg L −1 d −1 , respectively during the 30 °C (day)/16 °C (night) cycle. Saturated fatty acid content increased with increasing temperature. Additionally, we cultured Micractinium sp. under Nitrogen (N) and Phosphorus (P) limiting conditions. Highest lipid productivity of 85.4 ± 2 mg L −1 d −1 was obtained under P-depletion during exponential growth phase. Oleic acid amount also increased eight fold during P-deplete. [ABSTRACT FROM AUTHOR]

A novel mathematical model for the quantitative assessment of the effect of dissolved nitrogen on the autotrophic batch-growth and lipid accumulation of Chlorella sorokiniana , is proposed in this work. Model results have been validated through comparison with suitable experimental data performed in lab photobioreactors. Further experiments have been then performed using the BIOCOIL photobioreactor operated in fed-batch mode. The experimental results, which show that a maximum growth rate of 0.52 day −1 and a lipid content equal to 25 %wt can be achieved with the BIOICOIL, have been successfully predicted through the proposed model. Therefore, the model might represent a first step toward the development of a tool for the scale-up and optimization of the operating conditions of BIOCOIL photobioreactors. Finally, the fatty acid methyl esters obtained by trans-esterification of lipids extracted from C. sorokiniana , have been analyzed in view of the assessment of their usability for producing biodiesel. [ABSTRACT FROM AUTHOR]

Pure volatile fatty acid (VFA) solution derived from waste activated sludge (WAS) was used to produce microbial lipids as culture medium in this study, which aimed to realize the resource recovery of WAS and provide low-cost feedstock for biodiesel production simultaneously. Cryptococcus curvatus was selected among three oleaginous yeast to produce lipids with VFAs derived from WAS. In batch cultivation, lipid contents increased from 10.2% to 16.8% when carbon to nitrogen ratio increased from about 3.5 to 165 after removal of ammonia nitrogen by struvite precipitation. The lipid content further increased to 39.6% and the biomass increased from 1.56 g/L to 4.53 g/L after cultivation for five cycles using sequencing batch culture (SBC) strategy. The lipids produced from WAS-derived VFA solution contained nearly 50% of monounsaturated fatty acids, including palmitic acid, heptadecanoic acid, ginkgolic acid, stearic acid, oleic acid, and linoleic acid, which showed the adequacy of biodiesel production. [ABSTRACT FROM AUTHOR]

In the current study it has been hypothesized that, when organic loading of an anaerobic reactor is increased, the additional cell biomass biosynthesis would capture more ammonia nitrogen and thereby reduce the ammonia toxicity. Therefore, the alleviation of the toxicity of high ammonia levels using lipids (glycerol trioleate-GTO) or carbohydrates (glucose-GLU) as co-substrates in manure-based thermophilic continuous stirred-tank reactors (R GTO and R GLU , respectively) was tested. At 5 g NH 4 + -N L −1 , relative methane production of R GTO and R GLU , was 10.5% and 41% compared to the expected uninhibited production, respectively. At the same time control reactor (R CTL ), only fed with manure, reached 32.7% compared to the uninhibited basis production. Therefore, it seems that using lipids to counteract the ammonia effect in CSTR reactors creates an “ammonia–LCFA (long chain fatty acids) synergetic co-inhibition” effect. Moreover, co-digestion with glucose in R GLU was more robust to ammonia toxicity compared to R CTL . [ABSTRACT FROM AUTHOR]

To improve the biomass yield and lipid productivity, two desert microalgae, Desmodesmus sp. S81 and G41 were induced mutagenesis by Ethylmethane sulfonate (EMS), and obtained two potential mutants, Desmodesmus sp. S5 and G3 from the mutagenic clones for their greatly promoted biomass and lipid production. The results showed that the biomass yield, lipid content and lipid productivity of the mutant strains S5 and G3 were 778.10 mg·L −1 , 48.41% and 19.83 mg·L −1 ·d −1 , 739.52 mg·L −1 , 46.01%, and 17.92 mg·L −1 ·d −1 , respectively, which presented the increment of 45.50%, 8.00% and 74.24%, 20.67%, 10.35% and 55.77% than those of S81 and G41. Comparing with the wild strains, the mutants showed reduced PUFAs and glycol lipids, elevated MUFAs and neutral lipids contents, which were appropriate for biodiesel production. [ABSTRACT FROM AUTHOR]

In this study, crude glycerol from various industries was used to produce lipids via wild type Yarrowia lipolytica A101. We tested samples without any prior purification from five different waste products; each contained various concentrations of glycerol (42–87%) as the sole carbon source. The best results for lipid production were obtained for medium containing glycerol from fat saponification. This reached 1.69 g L −1 (25% of total cell dry weight) with a biomass yield of 0.17 g g −1 in the flasks experiment. The batch cultivation in a bioreactor resulted in enhanced lipid production—it achieved 4.72 g L −1 with a biomass yield 0.21 g g −1 . Moreover, the properly selected batch of crude glycerol provides a defined fatty acid composition. In summary, this paper shows that crude glycerol from soap production could be efficiently converted to single cell oil without any prior purification. [ABSTRACT FROM AUTHOR]

To improve nutrients removal from wastewater and enhance lipid production, cultivation of Chlorella vulgaris in wastewater with waste glycerol generated from biodiesel production using scum derived oil as feedstock was studied. The results showed that nutrients removal was improved and lipid production of C. vulgaris was enhanced with the addition of waste glycerol into wastewater to balance its C/N ratio. The optimal concentration of the pretreated glycerol for C. vulgaris was 10 g L −1 with biomass concentration of 2.92 g L −1 , lipid productivity of 163 mg L −1 d −1 , and the removal of 100% ammonia and 95% of total nitrogen. Alkaline conditions prompted cell growth and lipid accumulation of C. vulgaris while stimulating nutrients removal. The application of the integration process can lower both wastewater treatment and biofuel feedstock costs. [ABSTRACT FROM AUTHOR]

Biofuel production from microalgae requires energy efficient processes for extracting and converting triacylglyceride lipids to fuel, compatible with coproduction of protein feeds and nutraceuticals. Wet solvent extraction involves mechanical cell rupture, lipid extraction via solvent contacting, physical phase separation, thermal solvent recovery, and transesterification. A detailed analysis of the effect of key process parameters on the parasitic energy demand of this process was performed. On a well-to-pump basis, between 16% and 320% of the resultant biodiesel energy was consumed depending solely on the process parameters. Highly positive energy balances can be achieved, but only if a correctly designed process is used. This requires processing concentrated biomass ( ca 25% w/w) with a high triacylglyceride content ( ca 30% w/w), and an efficient extraction process employing a non-polar solvent, low solvent-to-paste ratio, and efficient energy recovery. These requirements preclude many laboratory scale processes and polar co-solvents as viable options for large-scale biofuel production. [ABSTRACT FROM AUTHOR]

Non-polar and polar solvents as well as their mixtures were tested for the extraction of microalgae lipids and thus, to evaluate their effect on total and esterifiable lipids extraction yields with potential to be converted to biodiesel. The obtained results show an increase in lipids and esterifiable lipids extraction yields when non-polar and polar solvent mixtures were used. The higher esterifiable lipids extraction yield was 19.2% wt (based on dry biomass) using a chloroform–methanol mixture (75% v/v of methanol), corresponding to a 98.9% wt esterifiable lipids extraction. In addition, esterifiable lipids extraction yield of 18.9% wt (based on dry biomass) was obtained when a petroleum ether–methanol mixture (75% v/v of methanol) was used, corresponding to a 96.9% wt esterifiable lipids extraction. [ABSTRACT FROM AUTHOR]

Kelp waste extracts (KWE) contained massive soluble sugars, amino acids and various mineral elements. To probe the effects of KWE on microalgal physiological and biochemical responses, the cultures were carried out under the different dilutions. The results showed that 8.0% KWE increased the biomass productivities and total lipid contents of Chlorella strains dramatically, which were 1.83–31.86 times and 20.78–25.91% higher than that of the control. Phaeodactylum tricornutum and Spirulina maxima presented a better growth performance in 1.0% and 4.0% treatment respectively, while their lipid accumulation were not enhanced. In Chlorella -Arc, Chlorella sorokiniana and P. tricornutum , the contents of saturated and monounsaturated fatty acids could be increased, and polyunsaturated fatty acids could be decreased under the conditions of high concentration of KWE (6.0–8.0%). Briefly, KWE facilitated to enhance the biomass productivity and lipid content of Chlorella strains, also improved the fatty acid compositions for biodiesel production. [ABSTRACT FROM AUTHOR]

Crude polysaccharides and proteins extracted from algae were chosen as model materials to investigate the hydrothermal liquefaction (HTL) characteristics and pathways of low-lipid algae. Liquefaction behavior of the two individuals and their binary mixtures with different mass ratios were evaluated under different temperatures. Formation pathways of bio-oil from crude polysaccharides/proteins were proposed. Results showed that polysaccharides had a small contribution to bio-oil (<5%) and approximately 60% distributed in aqueous phase, while proteins played a crucial role on bio-oil formation (maximum 16.29%). Bio-oil from polysaccharides mainly contained cyclic ketones and phenols and from proteins composed of pyrazines, pyrroles and amines. Interaction between polysaccharides and proteins forming polycyclic nitrogenous compounds had a negative effect on bio-oil yield at 220 and 260 °C. However, their further decomposition caused increase of bio-oil yield at 300 °C. Mixture liquefaction obtained the highest higher heating value (HHV) of bio-oil and energy recovery than polysaccharides/proteins liquefaction at 300 °C. [ABSTRACT FROM AUTHOR]

Chlorella protothecoides is a valuable source of lipids that may be used for biodiesel production. The present work shows analysis of the potential of photoheterotrophic cultivation of C. protothecoides under various light intensities aiming to identify the conditions with maximal biomass and lipid content. An increase in light intensity was associated with an increased specific growth rate and a shortened doubling time. Also, the relative total lipid content increased from 24.8% to 37.5% with increase of light intensity. The composition of fatty acid methyl esters was affected by light intensity with the C16–18 fatty acids increased from 76.97% to 90.24% of total fatty acids. However, the content of linolenic acids decreased with the increase of the culture irradiance. These studies indicate that cultures irradiated with high light intensities achieve the minimal specifications for biodiesel quality on linolenic acids and thus are suitable for biodiesel production. [ABSTRACT FROM AUTHOR]

This study presents gravimetric enrichment of mixed culture to screen starch and lipid producing species separately in a sequencing batch reactor. In the enriched starch-producing mixed culture photobioreactor, the starch content at the end of steady state batch became 3.42 times the beginning of depletion. Whereas in the enriched lipid-producing photobioreactor, the lipid content at the end of steady state batch became 3 times the beginning of famine phase. The obtained results revealed that the gravimetric enrichment is a suitable screening method for specific production of storage compounds in none-sterile large-scaled condition. [ABSTRACT FROM AUTHOR]

The development of a culture system for Chlorella sp. GD to efficiently produce biomass and oil for biodiesel production was investigated. Chlorella sp. GD was cultivated with 0%, 25%, 50%, 75% and 100% piggery wastewater (diluted by medium) at 300 μmol m −2 s −1 , a 2% CO 2 aeration rate of 0.2 vvm and 26 ± 1 °C; after a 10-day culture in batch cultures, the maximum specific growth rate and biomass productivity of the microalga obtained in 100% piggery wastewater were 0.839 d −1 and 0.681 g L −1 d −1 , respectively. The highest lipid content and lipid productivity were 29.3% and 0.155 g L −1 d −1 at 25% wastewater, respectively. In semi-continuous cultures, the biomass and lipid productivities with 25–75% wastewater ratios were greater than 0.852 and 0.128 g L −1 d −1 , respectively. These results show that Chlorella sp. GD grows efficiently in piggery wastewater, and that a stable growth performance was achieved for long-term microalgal cultivation in a semi-continuous culture. [ABSTRACT FROM AUTHOR]

A culture medium based on NPK-10:26:26 fertilizer was formulated for enhanced biomass and lipid production of Dunaliella tertiolecta by selecting appropriate nutrients and environmental parameters. Five-level-five-factor central composite design assisted response surface methodology was adopted for optimal cultivation of D. tertiolecta and results were compared with simple genetic algorithm (GA). Significant improvement in biomass and lipid production was obtained using newly formulated fertilizer medium over f /2 medium. Following optimal parameters [i.e., NaHCO 3 , (mM), NPK-10:26:26 (g L −1 ), NaCl (M), light intensity (μmol m −2 s −1 ) and temperature (°C)] were obtained for maximum biomass (1.98 g L −1 ) and lipid production (0.76 g L −1 ): (42.50, 0.33, 1.09, 125, 25.13) and (38.44, 0.40, 1.25, 125, 24.5), respectively using GA. A multi-objective optimization was solved using non-dominated sorting GA to find best operating variables to maximize biomass and lipid production simultaneously. Effects of operating parameters and their interactions on algae and lipid productivity were successfully revealed. [ABSTRACT FROM AUTHOR]

The pore structures and surface morphological characteristics of Nannochloropsis sp. cells with arsenic adsorption were initially investigated by N 2 -adsorption analysis and scanning electronic microscopy. Functional groups of cells were analysed by Fourier-transform infrared spectrometry and X-ray photoelectron spectroscopy. Total surface area of microalgal cells increased from 0.54 m 2 /g to 1.80 m 2 /g upon arsenic adsorption. The external cell surface area increased. More wrinkles and measles-like granules formed on the surfaces as a result of arsenic toxicity. Arsenic ions blocked cell pores and decreased the average pore diameter and total pore volume. Ether cross-linked structures in the algaenan layer of cell walls were disrupted as the percentage of C–O functional groups decreased. These functional groups underwent complexation reactions with arsenic ions. Accumulation of polyunsaturated fatty acids decreased because of oxidative stresses induced by arsenic. The increase in generation of short-chain saturated fatty acids was favourable for the production of quality biodiesel. [ABSTRACT FROM AUTHOR]

The marine eustigmatophyte Nannochloropsis is one of the potential producers of eicosapentaenoic acid (EPA), a valued nutraceutical. Nannochloropsis sp. was cultivated under photoautotrophic condition utilizing CO 2 in a two phase cultivation process in order to enhance the eicosapentaenoic acid (EPA) productivity. It was cultivated in a photobioreactor up to late log phase for cell growth (phase I). Then, the culture was harvested and confronted to relatively low temperature (10 °C) and low light (30 μmol photons m −2 s −1 ) in both photobioreactor and Erlenmeyer flask (phase II), thus augmenting EPA% by 3.4 fold. Lower temperature with low light favored the synthesis of EPA although, biomass productivity, lipid content and lipid productivity were slightly decreased relative to phase I. The total lipids extracted from Nannochloropsis sp. fractionated into neutral lipids (NLs), glycolipids (GLs) and phospholipids (PLs) and a major proportion of EPA was found in phospholipids. Results suggested that low temperature and low light may ameliorate partitioning towards EPA in phospholipids. [ABSTRACT FROM AUTHOR]

The aim of the present study was to find out the optimum sodium bicarbonate concentration to produce higher biomass with higher lipid and carbohydrate contents in microalgae Scenedesmus sp. CCNM 1077. The role of bicarbonate supplementation under different nutritional starvation conditions was also evaluated. The results clearly indicate that 0.6 g/L sodium bicarbonate was optimum concentration resulting in 20.91% total lipid and 25.56% carbohydrate along with 23% increase in biomass production compared to normal growth condition. Addition of sodium bicarbonate increased the activity of nutrient assimilatory enzymes, biomass, lipid and carbohydrate contents under different nutritional starvation conditions. Nitrogen starvation with bicarbonate supplementation resulted in 54.03% carbohydrate and 34.44% total lipid content in microalgae Scenedesmus sp. CCNM 1077. These findings show application of bicarbonate grown microalgae Scenedesmus sp. CCNM 1077 as a promising feedstock for biodiesel and bioethanol production. [ABSTRACT FROM AUTHOR]

A 2-kW continuous ultrasonic flow system (UFS) was found effective in the disruption of two microalgal strains: Scenedesmus dimorphus and Nannochloropsis oculata . Compared to the control, cell debris concentration of UFS treatments increased up to 202% for S. dimorphus and 112% for N. oculata. Similarly, Nile red stained lipid fluorescence density (NRSLD) increased up to 59.5% and 56.3% for S. dimorphus and N. oculata , respectively. It was also found that increasing ultrasound intensity improved cell disruption efficiency indicated by up to 54% increase in NRSLFD of the two strains. Increasing sonication-processing time to 3-min resulted in 33.0% increase for S. dimorphus and 45.7% increase for N. oculata in NRSLFD compared to the control. Cell recirculation was found beneficial to cell disruption, however, higher initial cell concentration significantly reduced cell disruption efficiency, indicated by 98.2% decrease in NRSLFD per cell when initial cell concentration increased from 4.25 × 10 6 to 1.7 × 10 7 cells ml −1 . [ABSTRACT FROM AUTHOR]

Microbial lipids produced by oleaginous yeasts serve as promising alternatives to traditional oils and fats for the production of biodiesel and oleochemicals. To improve its techno-economics, it is pivotal to use wastes and produce high quality lipids of special fatty acid composition. In the present study, four oleaginous yeasts were tested to use free fatty acids for lipid production under non-growth conditions. Microbial lipids of exceptionally high fatty acid relative contents, e.g. those contained over 70% myristic acid or 80% oleic acid, were produced that may be otherwise inaccessible by growing cells on various carbon sources. It was found that Cryptococcus curvatus is a robust strain that can efficiently use oleic acid as well as even-numbered saturated fatty acids with carbon atoms ranging from 10 to 20. Our results provided new opportunity for the production of functional lipids and for the exploitation of organic wastes rich in free fatty acids. [ABSTRACT FROM AUTHOR]

Extracting lipids from oleaginous microbial cells in a cost effective and environmentally compatible manner remains a critical challenge in developing manufacturing paradigms for advanced liquid biofuels. In this study, a new approach using microbial growth inhibitors from lignocellulose-derived feedstocks was used to extract lipids efficiently from wet cell mass of the oleaginous bacterium Rhodococcus opacus MITXM-61. Nine common lignocellulose-derived inhibitors for treatment of cells prior to solvent extraction were used and evaluated for their efficiency of lipid extraction from the cells. When the inhibitors were individually examined, formic acid and furfural showed the highest extraction efficiency of lipids from wet cell mass. Multiple extractions of lipids with methanol from wet cell mass pretreated with combined common inhibitors or hardwood hydrolysate comprising lignocellulose-derived inhibitors resulted in lipid recovery of greater than 85% of total lipids, a 1.7-fold increase of lipid extraction as compared to those in the absence of the inhibitors. [ABSTRACT FROM AUTHOR]

Chlorella vulgaris , a marine microalgae strain adaptable to 0–50 g L −1 of salinity, was selected for studying the coupling system of saline wastewater treatment and lipid accumulation. The effect of total nitrogen (TN) concentration was investigated on algal growth, nutrients removal as well as lipid accumulation. The removal efficiencies of TN and total phosphorus (TP) were found to be 92.2–96.6% and over 99%, respectively, after a batch cultivation of 20 days. To illustrate the response of lipid accumulation to nutrients removal, C. vulgaris was further cultivated in the recycling experiment of tidal saline water within the photobioreactor. The lipid accumulation was triggered upon the almost depletion of nitrate (<5 mg L −1 ), till the final highest lipid content of 40%. The nitrogen conversion in the sequence of nitrate, nitrite, and then to ammonium in the effluents was finally integrated with previous discussions on metabolic pathways of algal cell under nitrogen deficiency. [ABSTRACT FROM AUTHOR]

Methods to convert microalgal biomass to bio based fuels and chemicals are limited by several processing and economic hurdles. Research conducted in this study modified/optimized a previously published procedure capable of extracting transesterifiable lipids from wet algal biomass. This optimization resulted in the extraction of 77% of the total transesterifiable lipids, while reducing the amount of materials and temperature required in the procedure. In addition, characterization of side streams generated demonstrated that: (1) the C/N ratio of the residual biomass or lipid extracted (LE) biomass increased to 54.6 versus 10.1 for the original biomass, (2) the aqueous phase generated contains nitrogen, phosphorous, and carbon, and (3) the solid precipitate phase was composed of up to 11.2 wt% nitrogen (70% protein). The ability to isolate algal lipids and the possibility of utilizing generated side streams as products and/or feedstock material for downstream processes helps promote the algal biorefinery concept. [ABSTRACT FROM AUTHOR]

[Display omitted] • Lipid synthesis by oleaginous yeasts is a promising alternative to vegetable oils. • Y. lipolytica can bioconvert mixed C 2 -C 6 VFAs from gas fermentation into lipids. • Yarrowia lipolytica shows preference for acetic acid over butyric and caproic acids. • Y. lipolytica used the VFA mixture at a rate of 0.664 g/L·h with lipid accumulation. • The lipid profile obtained shows its high suitability for biodiesel production. Volatile fatty acids (VFAs) can be considered as low-cost carbon substrates for lipid accumulation by oleaginous yeasts. This study demonstrates that a common mixture of VFAs, typically obtained from the anaerobic fermentation of C1-gases by some acetogenic bacteria, can be used in a second aerobic fermentation with the yeast Yarrowia lipolytica to obtain lipids as precursors of biodiesel. In the batch experiments, the preference of Yarrowia lipolytica W29 for acetic acid over butyric and caproic acids was demonstrated, with the highest consumption rate reaching 0.664 g/L·h. In the bioreactor experiments, the amount initial biomass inoculated, as well as the initial acid concentration, were found to have a significant influence on the process. Though the lipid content was relatively low, it can be optimized and further improved. Oleic, linoleic and palmitic acids accounted for about 80 % of the fatty acids in the lipids, which makes them suitable for biodiesel. [ABSTRACT FROM AUTHOR]

[Display omitted] • Algal growth was boosted by various forms of sulfur with different dose dependent. • Effect of nitrogen on carbon assimilation in cells was regulated by sulfur form. • Sulfate was more beneficial to increase lipid productivity than other form. • Sulfur induced changes in expression of genes encoding lipid-related enzymes. Algal biodiesel has been becoming a focus in the field of bioenergy worldwide. In this study, effects of heterogeneous sulfur (SO 4 2-, SO 3 2- and S2-) on growth and lipid accumulation of Scenedesmus obliquus cultured in wastewater with a C/N ratio of 30 were investigated, respectively. The results shown that SO 4 2-, the optimal sulfur source, could trigger cell growth in a concentration-dependent manner. However, SO 3 2- was superior to the others in boosting carbon uptake of cells, which was subject to NH 4 +-N concentration. Only SO 4 2- could simultaneously increase lipid content and productivity of cells with a dominant component of oleic acid (C18:1n9c) occupying approximately 40% in fatty acid profile. Additionally, the genes encoding enzymes such as CDIPT, ADPRM, DPP1, pmtA and BTA1 involved in the uppermost lipid-related pathway (glycerophospholipid metabolism) were identified facing different sulfur source regardless of the concentration changes. These findings may facilitate nutrition management efforts to enhance microalgae-based biofuel production. [ABSTRACT FROM AUTHOR]

[Display omitted] • Chlorella sorokiniana cultivation under photoautotrophic or heterotrophic conditions. • Higher lipids (3.19 ± 0.23 g/L) at C/N 60 after 72 h growth on heterotrophic mode. • Autotrophic cultivation favored proteins (416.66 mg/g CDW) & lutein (6.40 mg/g CDW) • Total Carbohydrates for photoautotrophic (6.39 %) and heterotrophic (8.66 %) modes. • The lipid profile was suitable for its application in biodiesel production. Biorefineries enable the circular, sustainable, and economic use of waste resources if value-added products can be recovered from all the generated fractions at a large-scale. In the present studies the comparison and assessment for the production of value-added compounds (e.g., proteins, lutein, and lipids) by the microalga Chlorella sorokiniana grown under photoautotrophic or heterotrophic conditions was performed. Photoautotrophic cultivation generated little biomass and lipids, but abundant proteins (416.66 mg/g CDW) and lutein (6.40 mg/g CDW). Heterotrophic conditions using spruce hydrolysate as a carbon source favored biomass (8.71 g/L at C/N 20 and 8.28 g/L at C/N 60) and lipid synthesis (2.79 g/L at C/N 20 and 3.61 g/L at C/N 60) after 72 h of cultivation. Therefore, heterotrophic cultivation of microalgae using spruce hydrolysate instead of glucose offers a suitable biorefinery concept at large-scale for biodiesel-grade lipids production, whereas photoautotrophic bioreactors are recommended for sustainable protein and lutein biosynthesis. [ABSTRACT FROM AUTHOR]

[Display omitted] • Nordic microalgae produce compounds of high value when exposed to abiotic stress. • Light stress stimulates carotenoid production up to 44 mg L−1 in Desmodesmus sp. • Combined light and salt stress enhanced the lipid content by 43% of DW. • Nordic microalgae grown in various wastewaters can produce carotenoids and lipids. Nordic Desmodesmus microalgal strains (2–6) and (RUC-2) were exposed to abiotic stress (light and salt) to enhance lipids and carotenoids. The biomass output of both strains increased by more than 50% during light stress of 800 μmol m−2 s−1 compared to control light. The biomass of Desmodesmus sp. (2–6) contained most lipids (15% of dry weight) and total carotenoids (16.6 mg g−1) when grown at moderate light stress (400 μmol m−2 s−1), which further could be enhanced up to 2.5-fold by salinity stress. Desmodesmus sp. (RUC-2) exhibited maximal lipid (26.5%) and carotenoid (43.8 mg L−1) content at light intensities of 400 and 100 μmol m−2 s−1, respectively. Salinity stress stimulated lipid accumulation by 39%. Nordic Desmodesmus strains therefore are not only able to tolerate stress conditions, but their biomass considerably improves under stress. These strains have high potential to be used in algal bio-factories on low-cost medium like Baltic seawater. [ABSTRACT FROM AUTHOR]

[Display omitted] • The combined application of SA and NaSH heightened astaxanthin and lipid yields. • SA and NaSH induction upregulated growth and carotenogenesis gene expression levels. • Cellular H 2 S mediated ROS signal transduction and enzymatic antioxidant levels. To obtain higher yield of natural astaxanthin, the present study aims to develop a viable and economic induction strategy for astaxanthin production comprising succinic acid (SA) combined with sodium hydrosulfide (NaHS). The biomass (1.33 g L-1), astaxanthin concentration (44.96 mg L-1), astaxanthin content (163.55 pg cell−1), and lipid content (55.34%) were achieved under 1.0 mM SA and 100 μM NaHS treatment. These results were concomitant with enhanced hydrogen sulfide (H 2 S) but diminished reactive oxide species (ROS). Further study discovered that endogenous H 2 S could improve astaxanthin and lipid coproduction under SA induction by mediating related gene transcript levels and ROS signalling. Additionally, the concentrations of biomass and astaxanthin increased to 2.14 g L-1 and 66.25 mg L-1, respectively, under the induction of SA and NaHS in a scaled-up bioreactor. Briefly, the work proposed a novel feasible strategy for high yields of biomass and astaxanthin by H. pluvialis. [ABSTRACT FROM AUTHOR]

This work investigated a top-lit open microalgae bioreactor that uses a gas-lift system to enable deeper production depths, thereby significantly reducing the footprint. Growth of Scenedesmus sp. in a one-meter deep system by sparged with 6% CO 2 -enhanced air was evaluated. The results gave comparable volumetric biomass productivity (0.06 g dw L −1 day −1 ), but around three-times higher areal productivity (60.0 g dw m − 2 day − 1 ) than reported for traditional raceways. The lipid content of the Scenedesmus sp. was increased by 27% with an enhanced level of CO 2 in the sparging gas. [ABSTRACT FROM AUTHOR]

A local Chlorella sp . isolate with 97% rbcL sequence identity to Chlorella sorokiniana was evaluated in terms of its biomass and lipid production under mixotrophic growth conditions. Glucose-supplemented cultures exhibited increasing growth rate and biomass yield with increasing glucose concentration. Highest growth rate and biomass yield of 1.602 day −1 and 687.5 mg L −1 , respectively, were achieved under 2 g L −1 glucose. Nitrogen starvation up to 75% in the 1.0 g L −1 glucose-supplemented culture was done to induce lipid accumulation and did not significantly affect the growth. Lipid content ranges from 20% to 27% dry weight. Nile Red staining showed more prominent neutral lipid bodies in starved mixotrophic cultures. C. sorokiniana exhibited enhanced biomass production under mixotrophy and more prominent neutral lipid accumulation under nitrogen starvation with no significant decrease in growth; hence, this isolate could be further studied to establish its potential for biodiesel production. [ABSTRACT FROM AUTHOR]

In this work, neutral lipids (NLs) extraction capacity and selectivity of six solvents were firstly compared. In addition, an eco-friendly solvent combination of ethyl acetate and ethanol (EA/E) was proposed and tested for lipid extraction from Chlorella sp. cultivated in outdoor raceway ponds and effect of extraction variables on lipid yield were intensively studied. Results indicated that lipid extraction yield was increased with solvent to biomass ratio but did not vary significantly when the value exceeded 20:1. Lipid yield was found to be strongly dependent on extraction temperature and time. Finally, fatty acid profiles of lipid were determined and results indicated that the major components were octadecanoic acid, palmitic acid, linoleic acid and linolenic acid, demonstrating that the lipid extracted from the Chlorella sp. cultivated in outdoor raceway ponds by EA/E was suitable feedstock for biodiesel production. [ABSTRACT FROM AUTHOR]

In this study, different light intensities (80, 160, 240 and 320 μmol/m 2 s) and various mediums including control medium (CM), N/P rich medium (NPM), N rich medium (NM), and P rich medium (PM) were applied for cultivation of Chlorella sp. It was revealed that cultivation of Chlorella sp. in CM under the light intensity of 320 μmol/m 2 s led to a lipid content up to 30% enhancement, which was higher than the results of other cases. A rather high unsaturated fatty acid (UFA) content of 7.5% and unsaturated fatty acid/total fatty acid (UFA/TFA) ratio of 0.73 were obtained under 320 μmol/m 2 s in CM, indicating that the CM-320 system was applicable for the generation of UFA. Moreover, Chlorella sp. cultivated in PM under 320 μmol/m 2 s provided higher TFA content (7.3%), which was appropriate for biofuel production. [ABSTRACT FROM AUTHOR]

The aim of this study was to select a potential microalgal strain for lipid production and to examine the suitability of anaerobically treated piggery wastewater as a nutrient source for production of lipid-rich biomass with the selected microalga. Biomass and lipid productivity of three microalgal strains ( Chlorella sorokiniana CY1, Chlorella vulgaris CY5 and Chlamydomonas sp . JSC-04) were compared by using different media, nitrogen sources, and nitrogen concentrations. The highest lipid content and productivity (62.5 wt%, 162 mg/L/d) were obtained with C. vulgaris with BG-11 with 62 mg N/L. Secondly, C. vulgaris was cultivated in sterilized, diluted (1–20×), anaerobically treated piggery wastewater. Biomass production decreased and lipid content increased, when wastewater was more diluted. The highest lipid content of 54.7 wt% was obtained with 20× dilution, while the highest lipid productivity of 100.7 mg/L/d with 5× dilution. Piggery wastewater is a promising resource for mass production of oleaginous microalgal biomass. [ABSTRACT FROM AUTHOR]

In this study, pyrite (FeS 2 ) was used for lipid extraction as well as esterification processes for microalgae-based biodiesel production. An iron-mediated oxidation reaction, Fenton-like reaction, produced an expected degree of lipid extraction, but pyrite was less effective than FeCl 3 commercial powder. That low efficiency was improved by using oxidized pyrite, which showed an equivalent lipid extraction efficiency to FeCl 3 , about 90%, when 20 mM of catalyst was used. Oxidized pyrite was also employed in the esterification step, and converted free fatty acids to fatty acid methyl esters under acidic conditions; thus, the fatal problem of saponification during esterification with alkaline catalysts was avoided, and esterification efficiency over 90% was obtained. This study clearly showed that pyrite could be utilized as a cheap catalyst in the lipid extraction and esterification steps for microalgae-based biodiesel production. [ABSTRACT FROM AUTHOR]

Instant catapult steam explosion (ICSE) was employed to disrupt wet microalgal cells for efficient lipid extraction. Physicochemical properties of exploded cells were investigated through SEM, TEM, FTIR, and TGA. The exploded cells increased in fractal dimension (1.53–1.65) when preheat time was prolonged from 0 min to 5 min and in surface pore area when steam pressure was increased. Meanwhile, the exploded cells decreased in mean size (1.69–1.44 μm) when the filling ratio of wet microalgal biomass in the preheat chamber decreased (75–12.5%). Flash evaporation and volume expansion exploded the cell walls and released the cytoplasm of the microalgal cells. These phenomena decreased the carbohydrate content and increased the lipid content in the exploded biomass. However, ICSE treatment did not change the lipid compositions in the microalgal cells. Using isopropanol as a cosolvent significantly increased the yield of lipids extracted with hexane from the exploded wet microalgal biomass. [ABSTRACT FROM AUTHOR]

Bio-based fuels are becoming more and more important due to the depleting fossil resources. The production of biodiesel from algae oil is challenging compared to terrestrial vegetable oils, as algae oil consists of polar fatty acids, such as phospholipids and glycolipids, as well as non-polar triglycerides and free fatty acids common in vegetable oils. It is shown that a single sulphonated solid acid catalyst can perform the esterification and transesterification reactions of both polar and non-polar lipids. In mild reaction conditions (60–70 °C) Nafion NR50 catalyst produces methyl palmitate (FAME) from the palmitic acid derivatives of di-, and tri-glyceride, free fatty acid, and phospholipid with over 80% yields, with the glycolipid derivative giving nearly 40% yields of FAME. These results demonstrate how the polar and non-polar lipid derivatives of algal oil can be utilised as feedstocks for biodiesel production with a single catalyst in one reaction step. [ABSTRACT FROM AUTHOR]

Hydrothermal acid treatment, was adopted to extract eicosapentaenoic acid (EPA) from wet biomass of Nannochloropsis salina . It was found that sulfuric acid-based treatment increased EPA yield from 11.8 to 58.1 mg/g cell in a way that was nearly proportional to its concentration. Nitric acid exhibited the same pattern at low concentrations, but unlike sulfuric acid its effectiveness unexpectedly dropped from 0.5% to 2.0%. The optimal and minimal conditions for hydrothermal acid pretreatment were determined using a statistical approach; its maximum EPA yield (predicted: 43.69 mg/g cell; experimental: 43.93 mg/g cell) was established at a condition of 1.27% of sulfuric acid, 113.34 °C of temperature, and 36.71 min of reaction time. Our work demonstrated that the acid-catalyzed cell disruption, accompanied by heat, can be one potentially promising option for ω-3 fatty acids extraction. [ABSTRACT FROM AUTHOR]

In the study, fourteen chemical modulators from five groups ( i.e. , auxin, gibberellin, cytokinin, signal transducer and amine) were evaluated for their effects on lipid accumulation in Crypthecodinium cohnii . The results showed that naphthoxyacetic acid (BNOA), 2-chlorodracylicacid, salicylic acid (SA), abscisic acid (ABA) and ethanolamine (ETA), increased lipid accumulation in C. cohnii by 10.00–18.78%. In addition, the combined uses of the above chemicals showed that two combinations, 1.0 mg/L SA & 152.7 mg/L ETA and 4.0 mg/L BNOA & 152.7 mg/L ETA, increased lipid accumulation by 22.45% and 20.54%, respectively. Moreover, a targeted metabolomic approach was employed to decipher the possible mechanisms responsible for the increased lipid accumulation, and the results showed that the enhanced metabolism in glycolysis and TCA cycle as well as the decreased metabolism in PPP pathway could be important for the stimulatory roles of BNOA & ETA and SA & ETA on lipid accumulation in C. cohnii . [ABSTRACT FROM AUTHOR]

The fermentation process for high yields of fatty acid and neutral lipid production from cassava bagasse hydrolysate (CBH) was developed by heterotrophic Chlorella protothecoides . An efficient single-step enzymatic hydrolysis of cassava bagasse (CB) by cellulase was firstly developed to produce >30 g/L of reducing sugars. The concentrated CBH was subsequently applied in a batch culture, producing 7.9 g/L of dry biomass with yield of 0.44 g/g reducing sugar and 34.3 wt% of fatty acids and 48.6 wt% of neutral lipids. Furthermore, fed-batch fermentation using CBH achieved higher yields of fatty acids (41.0 wt% and a titer of 5.83 g/L) and neutral lipids (58.4 wt% and yield of 0.22 g/g reducing sugar). Additionally, the fatty acid profile analysis showed that the intercellular lipid was suitable to prepare high-quality biodiesel. This study demonstrated the feasibility of using CBH as low-cost feedstock to produce crude algal oil for sustainable biodiesel production. [ABSTRACT FROM AUTHOR]

This work aimed to find an optimum culture medium for green microalga Botryococcus braunii KMITL and investigate its biodiesel properties based on fatty acid composition. Four different media were tested. Chlorella medium was the best medium for lipid yield. Among four nitrogen sources tested, KNO 3 produced the highest lipid yield. When varied the nitrogen concentrations, this strain gave the highest lipid yield at the highest nitrogen level. When cultivated in the best medium and nitrogen source and level for 30 days, and then cultivated further for 14 days in the medium with no nitrogen, the highest lipid content and yield were 49.94 ± 0.82% and 2.71 ± 0.02 g L −1 , respectively. C16:0 fatty acid was the major fatty acid found. Fatty acid profiles of B. braunii KMITL cultivated in Chlorella medium with 1.25 g L −1 KNO 3 gave the best biodiesel properties with the lowest iodine value, maximum cetane number, and lowest degree of unsaturation. [ABSTRACT FROM AUTHOR]

Critical influence of different short chain fatty acids as organic carbon source, during growth (GP) and nutrient stress lipogenic phase (NSLP) was investigated on biomass and lipid productivity, in mixotrophic fed-batch microalgae cultivation. Nutrient deprivation induced physiological stress stimulated highest lipid productivity with acetate (total/neutral lipids, 35/17) with saturation index of 80.53% by the end of NSLP followed by butyrate (12/7%; 78%). Biomass growth followed the order of acetate (2.23 g/l) >butyrate (0.99 g/l) >propionate (0.77 g/l). VFA removal (as COD) was maximum with acetate (87%) followed by butyrate (55.09%) and propionate (10.60%). Palmitic acid was the most dominant fatty acid found in the fatty acid composition of all variants and butyrate fed system yielded a maximum of 44% palmitic acid. Protein profiling illustrated prominence of acetyl CoA-synthetase activity in acetate system. Thus, fatty acids provide a promising alternative feedstock for biodiesel production with integrated microalgae-biorefinery. [ABSTRACT FROM AUTHOR]

Novel strategy for enhancing TAG accumulation by simultaneous utilization of fermentable and non-fermentable carbon sources as substrate for cultivation of oleaginous yeast Rhodosporidium kratochvilovae HIMPA1 were undertaken in this investigation. The yeast strain showed direct correlation between the size of lipid bodies, visualized by BODIPY stain (493–515 nm) and TAG accumulation when examined on individual fermenting and non-fermenting carbon sources and their mixtures. Maximum TAG accumulation (μm) in glucose (2.38 ± 0.52), fructose (4.03 ± 0.38), sucrose (4.24 ± 0.45), glycerol (4.35 ± 0.54), xylulose (3.94 ± 0.12), and arabinose (2.98 ± 0.43) were observed. Synergistic effect of the above carbon sources (fermentable and non-fermentable) in equimolar concentration revealed maximum lipid droplet size of 5.35 ± 0.76 μm and cell size of 6.89 ± 0.97 μm. Total lipid content observed in mixed carbon sources was 9.26 g/l compared to glucose (6.2 g/l). FAME profile revealed enhanced longer chain (C 14:0 -C 24:0 ) fatty acids in mix carbon sources. [ABSTRACT FROM AUTHOR]