In order to comply with criteria of green chemistry concepts and sustainability, a new procedure has been performed for solvent-free ultrasound-assisted extraction (UAE) to extract lipids from fresh Nannochloropsis oculata biomass. Through response surface methodology (RSM) parameters affecting the oil recovery were optimized. Optimum conditions for oil extraction were estimated as follows: (i) 1000 W ultrasonic power, (ii) 30 min extraction time and (iii) biomass dry weight content at 5%. Yields were calculated by the total fatty acids methyl esters amounts analyzed by GC-FID-MS. The maximum oil recovery was around 0.21%. This value was compared with the one obtained with the conventional extraction method (Bligh and Dyer). Furthermore, effect of temperature on the yield was also investigated. The overall results show an innovative and effective extraction method adapted for microalgae oil recovery, without using solvent and with an enable scaling up.
Nannochloropsis sp. F&M-M24 and Tetraselmis suecica F&M-M33 were cultivated outdoors in Green Wall Panels under nutrient deficiency to stimulate oil synthesis. Under nitrogen deprivation, Nannochloropsis attained average biomass and lipid productivities of 9.9 and 6.5 g m(-2) day(-1), respectively. Starved Tetraselmis cultures achieved a biomass productivity of about 7.6 g m(-2) day(-1) and a lipid productivity of 1.7 g m(-2) day(-1). Lipids represented 39.1% and 68.5% of non-starved and starved Nannochloropsis biomass, respectively. Starvation did not increase lipid content in Tetraselmis biomass. Important differences in lipid classes and in fatty acid composition were observed under the different cultivation conditions for both microalgae.
Magnetic collection of the microalgae Chlorella sp. from culture media facilitated by low-gradient magnetophoretic separation is achieved in real time. A removal efficiency as high as 99% is accomplished by binding of iron oxide nanoparticles (NPs) to microalgal cells in the presence of the cationic polyelectrolyte poly(diallyldimethylammonium chloride) (PDDA) as a binder and subsequently subjecting the mixture to a NdFeB permanent magnet with surface magnetic field ≈6000 G and magnetic field gradient <80 T m(-1) . Surface functionalization of magnetic NPs with PDDA before exposure to Chlorella sp. is proven to be more effective in promoting higher magnetophoretic removal efficiency than the conventional procedure, in which premixing of microalgal cells with binder is carried out before the addition of NPs. Rodlike NPs are a superior candidate for enhancing the magnetophoretic separation compared to spherical NPs due to their stable magnetic moment that originates from shape anisotropy and the tendency to form large NP aggregates. Cell chaining is observed for nanorod-tagged Chlorella sp. which eventually fosters the formation of elongated cell clusters.
Bio-sequestration of CO(2) through microalgae cultivation is considered as a viable option for biofuel production as well as CO(2) mitigation. Influence of CO(2) sparging period and interval was evaluated on the growth and lipid accumulation of microalgae cultivated in domestic wastewater under mixotrophic microenvironment. Process performance was assessed in two phases viz., growth (GP) and starvation phases (SP) each with 8 days of retention time. Experimental variations depicted marked influence on biomass growth and lipid accumulation of microalgae with the function of harvesting period. Sparging period of 120 s documented maximum biomass growth (GP, 3.4 mg/ml) and lipid productivity (SP, 27.3%) while in intervals, 4h (120 s) condition showed maximum biomass (3.2mg/ml) and lipid productivity (27.8%). Total chlorophyll components documented higher concentrations of Chl b supporting the observed higher lipid productivity. Fatty acid composition varied with the experimental variations and represented higher degree of saturation indicating their utility as biodiesel.
Microalgae-based bioenergy has gained extensive attention, but the consumption of non-renewable resource such as phosphorous is inevitable in the production of its feedstock. In this work, the minimal phosphorous consumption for algal biomass production of Scenedesmus sp. LX1 was investigated by monitoring the growth and nutrient uptake under two different cultivation modes: phosphorous-starvation and luxury-nutrient. The results showed that continuous nitrogen and phosphorous feeding in luxury-nutrient mode had no stimulating effect on biomass productivity at the nutrient level in this study, TN: 245 mg L(-1), TP: 5.4 mg L(-1). However, the sustained growth of biomass after the exhaust of phosphate in phosphorous-starvation mode led to significant increase in the biomass yield of phosphorous up to 160 g biomass/g -P, which was nearly six times more than that with nutrient feeding. To minimize phosphorous resource consumption in production of algal biomass, a phosphorous-starvation cultivation mode is proposed.
In an effort to search for an efficient and environmentally friendly harvesting method, a commercially available microbial flocculant poly (γ-glutamic acid) (γ-PGA) was used to harvest oleaginous microalgae. Conditions for flocculation of marine Chlorella vulgaris and freshwater Chlorella protothecoides were optimized by response surface methodology (RSM) and determined to be 22.03 mg L(-1) γ-PGA, 0.57 g L(-1) biomass, and 11.56 g L(-1) salinity, and 19.82 mg L(-1) γ-PGA and 0.60 g L(-1) biomass, respectively. Application of the two optimized flocculation methods to Nannochloropsis oculata LICME 002, Phaeodactylum tricornutum, C. vulgaris LICME 001, and Botryococcus braunii LICME 003 gave no less than 90% flocculation efficiency and a concentration factor greater than 20. Micrographs of the harvested microalgal cells showed no damage to cell integrity, and hence no lipid loss during the process. The results show that flocculation with γ-PGA is feasible for harvesting microalgae for biodiesel production.
Flue gases are a resource yet to be fully utilised in microalgal biotechnology, not only to moderate the anthropogenic effects on our climate, but also to steer microalgal resource management towards innovative applications of microalgal biomass compounds. These gases, both untreated and treated into current discharge standards, contain CO(2), N(2), H(2)O, O(2), NO(x), SO(x), C(x)H(y), CO, particulate matter, halogen acids and heavy metals. To better steer and engineer flue gas-fed microalgal cultures, all these compounds need to be considered. Therefore, here, we review (i) the chemical composition and treatment technologies of flue gas, (ii) the uptake pathways and removal of the different compounds in microalgae reactors, and (iii) the tolerance and effects on microalgae of all flue gas compounds. By emphasising the interactions between microalgae and flue gas compounds, we envisage new pathways for microalgal biomass valorisation such as enzyme production for environmental technology, novel biogas production and biosequestration of minerals. Furthermore, we highlight fundamental and applied research niches that merit further investigation.
Two human clinical studies were performed. One was an open-label non-controlled study involving 30 healthy female subjects for 8 weeks. Significant improvements were observed by combining 6 mg per day oral supplementation and 2 ml (78.9 μM solution) per day topical application of astaxanthin. Astaxanthin derived from the microalgae, Haematococcus pluvialis showed improvements in skin wrinkle (crow's feet at week-8), age spot size (cheek at week-8), elasticity (crow's feet at week-8), skin texture (cheek at week-4), moisture content of corneocyte layer (cheek in 10 dry skin subjects at week-8) and corneocyte condition (cheek at week-8). It may suggest that astaxanthin derived from H. pluvialis can improve skin condition in all layers such as corneocyte layer, epidermis, basal layer and dermis by combining oral supplementation and topical treatment. Another was a randomized double-blind placebo controlled study involving 36 healthy male subjects for 6 weeks. Crow's feet wrinkle and elasticity; and transepidermal water loss (TEWL) were improved after 6 mg of astaxanthin (the same as former study) daily supplementation. Moisture content and sebum oil level at the cheek zone showed strong tendencies for improvement. These results suggest that astaxanthin derived from Haematococcus pluvialis may improve the skin condition in not only in women but also in men.
Triacylglycerols (TAGs) from microalgae have the potential to be used for biodiesel, but several technical and economic hurdles have to be overcome. A major challenge is efficient extraction of intracellular TAGs from algae. Here we investigate the use of enzymes to deconstruct algal cell walls/membranes. We describe a rapid and simple assay that can assess the efficacy of different enzyme treatments on TAG-containing algae. By this means crude papain and bromelain were found to be effective in releasing TAGs from the diatom Phaeodactylum tricornutum, most likely because of their cysteine protease activity. Pre-treating algal biomass with crude papain enabled complete extraction of TAGs using heptane/isopropyl alcohol. Heptane as a single solvent was also effective, although complete recovery of TAG was not obtained. Economic implications of these findings are discussed, with the aim to reduce the complexity of, and energy needed in, TAG extraction.
Application of photosynthetic micro-organisms, such as cyanobacteria and green algae, for the carbon neutral energy production raises the need for cost-efficient photobiological processes. Optimization of these processes requires permanent control of many independent and mutably dependent parameters, for which a continuous cultivation approach has significant advantages. As central factors like the cell density can be kept constant by turbidostatic control, light intensity and iron content with its strong impact on productivity can be optimized. Both are key parameters due to their strong dependence on photosynthetic activity.
Here we introduce an engineered low-cost 5 L flat-plate photobioreactor in combination with a simple and efficient optimization procedure for continuous photo-cultivation of microalgae. Based on direct determination of the growth rate at constant cell densities and the continuous measurement of O2 evolution, stress conditions and their effect on the photosynthetic productivity can be directly observed.
As a consequence of increased insulation levels, the growth of algae on façades has developed into a serious aesthetic problem over the past decade. Manufacturers of façade and coating materials are looking for strategies to avoid or at least delay the establishment of biofilms. Efficient product development, however, is complicated by the necessity of time-consuming free-weathering tests and a lack of objective measures to quantify the growth. In a two-year study the potential of pulse-amplitude modulation (PAM) fluorometry was tested for the quantification of algal growth on free-weathered specimens below the visual threshold. By using an imaging PAM in combination with a scanning device, surface colonization by algae was repeatedly visualized and analysed in detail. It was demonstrated that in the case of water saturation the resistance of a specific paint against algal growth can be quantified with sufficient accuracy by measuring the fluorescence yield after dark adaptation. The representative growth pattern identified, however, is not characterized by a linear accumulation of algal colonies but by a long phase of fluctuating biomass at a low level, followed by a steep increase. Only if the fluorescence yield rises significantly above the basic value does the specimen simultaneously show green discoloration. The duration of free-weathering tests, therefore, cannot be significantly shortened by using the more sensitive diagnostic tool.
A green microalgae strain, Chlorococcum sp., was obtained from tropical freshwater in Indonesia. The effects of pH on growth, effects of salt, carbon dioxide level, nitrate, acetate, and photoperiod on biomass and lipid production were studied. The fatty acids composition was also investigated. This study shows that the strain had an optimum pH value for growth ranging from pH 8.0 to 8.5. The biomass concentration and lipid content were influenced by different concentrations of salt, CO2 and nitrate. The lipid productivity ranged from 2 to 90.8 mg L-1 d-1 in different mediums. The highest biomass concentration and total lipid content achieved were 1.75 g L-1 and 56% of dry weight, respectively. Moreover, the major fatty acid methyl esters were C16:0, C18:1, C18:2 and C18:3. The high lipid content and the fatty acid composition make the strain Chlorococcum sp. a potential resource for food, cosmetics and biodiesel.
This study evaluated the use of natural coagulants (Moringa oleifera and chitosan) under different conditions with a mixed culture (C1 mixed culture). This culture was used for the biodegradation of hydrocarbons present in the effluent from fuel distribution terminals contaminated with diesel oil and gasoline. The biodegradation was evaluated by two central composite design (CCD) experiments: the first with varying concentrations of Moringa oleifera (MO), drying temperatures (TE) and seed drying times (TI); the second with varying concentrations of chitosan and the hydrochloric acid in which chitosan had been solubilized. The responses monitored in the CCD experiments included the sludge volume index (SVI), the turbidity removal (TR) and the specific rate of oxygen uptake (SOUR). Subsequently, the biodegradation was monitored in a sequencing batch reactor (SBR) under the optimal conditions obtained for each CCD experiment. The results indicated that the best coagulant was chitosan solubilized in 0.25 N HCl at a concentration of 50 mg/L. Within five cycles with chitosan as a coagulant, the total organic carbon (TOC) removal increased from 77 ± 1.0% to 82 ± 0.5%, the volatile suspended solids (VSS) increased from 1.4 ± 0.3 to 2.25 ± 0.3 g/L and the total petroleum hydrocarbon (TPH) removal increased from 75 ± 1.0% to 81 ± 0.5%.
Nitrogen availability and light intensity affect β-carotene overproduction in the green alga Dunaliella salina. Following a previous study on high-light stress, we here report on the effect of nitrogen depletion on the growth characteristics and β-carotene as well as fatty acid metabolism of D. salina under a constant light regime in a turbidostat. Upon nitrogen depletion, the biomass yield on absorbed light approximately doubled, due to a transient increase in cell division rate, swelling of the cells and a linear increase of the density of the cells. Simultaneously, β-carotene started to accumulate up to a final intracellular concentration of 14 mg LCV−1 (i.e. 2.7% of AFDW). This β-carotene production accounted for 6% of the increased density of the cells, indicating that other biochemical constituents accumulated as well. Since D. salina accumulates β-carotene in lipid globules, we also determined the fatty acid content and composition of D. salina. The intracellular concentration of the total fatty acid pool did not change significantly during nitrogen starvation, indicating that β-carotene and total fatty acid accumulation were unrelated, similar to what was found previously for high-light treated cells. However, for both high-light and nitrogen stress, β-carotene accumulation negatively correlated with the degree of unsaturation of the total fatty acid pool and, within the individual fatty acids, correlated positively with oleic acid biosynthesis, suggesting that oleic acid may be a key component of the lipid-globule-localized triacylglycerols and thereby in β-carotene accumulation.
Carbon dioxide sequestration using microalgae is the most promising method for combating global warming. Growth of microalgae is influenced by the availability of carbon dioxide, number of photons, initial concentration of microalgae and nutrients. The transfer of carbon dioxide from flue gas and absorption of photons from sunlight are influenced by the surface area/volume ratio of photobioreactor. The growth rate of microalgae follows lag, log, deceleration and stationary phases. The rate of growth increases with concentration of microalgae till an optimum concentration of algae is reached and then decreases for any fixed operating conditions and selected microalgae. At an optimum concentration the rate is the highest always. Operating a photobioreactor at this optimum concentration with highest surface area to volume ratio would require the smallest size of photobioreactor for a given production rate. Based on the review on the performance of various existing photobioreactors and the growth mechanism of microalgae it is observed that the design and operation of an efficient photo bioreactor system should consider (1) providing highest spread area to volume ratio (2) maintaining optimum concentration matching the highest rate (3) harvesting the excess microalgae formed over the optimum concentration to maintain the optimum concentration and (4) adding nutrients to the growth medium to maintain nutrient concentration at a constant level.
Haematococcus pluvialis synthesizes a high yield of astaxanthin using CO2 in a photoautotrophic culture without contaminant heterotrophs; however, it takes too long to induce astaxanthin production. In this study, a highly-photosensitive mutant strain was attained by conventional random mutagenesis and an efficient isolation method to shorten induction time. Sensitivity to photoinhibition in this mutant was raised by a partial lesion in the photosystem II (PSII) of photosynthesis, thereby prompting a change in cellular morphology as well as stimulating carotenogenesis (astaxanthin production). As a result, the concentrations of cell biomass and astaxanthin were dramatically increased by 27% and 62% under strong light and 79% and 153% under moderate light, respectively. This Haematococcus mutant would be useful for the economical astaxanthin production capable of reducing the light energy cost in a photoautotrophic culture system, even in areas with insufficient sunlight.
This study focuses on a scale-up procedure considering two vital parameters light energy and mixing for microalgae cultivation, taking Chlamydomonas reinhardtii as the model microorganism. Applying two stage hydrogen production protocol to 1L flat type and 2,5L tank type photobioreactors hydrogen production was investigated with constant light energy and mixing time. The conditions that provide the shortest transfer time to anaerobic culture (light energy; 2,96 kJ s-1 m-3 and mixing time; 1 min) and highest hydrogen production rate (light energy; 1,22 kJ s-1 m-3 and mixing time; 2,5 min) are applied to 5L photobioreactor. The final hydrogen production for 5L system after 192 hours was measured as 195±10 mL that is comparable with the other systems is a good validation for the scale-up procedure.
The conversion of microalgae lipids to biodiesel has been increasingly investigated, with the use of the in-situ transesterification method reported in the literature to lead to improved alkyl ester conversions for this feedstock compared with the use of the conventional two staged oil extraction and transesterification process. To further improve the feasibility of the use of the in-situ method, this paper investigates modifications to reduce the large process methanol requirements, and potentially improve the oil to methyl esters conversion and biodiesel yields. The results obtained showed that use of ultrasound agitation for the in-situ process, as well as combining this stirring regime with co-solvent use (n-pentane and diethyl ether) significantly improved the Chlorella oil to methyl esters conversion with reduced reacting methanol volumes.
Microalgae have been suggested as a promising feedstock for producing biofuel because of their potential of lipid production. In this study, a first principles ODE model for microalgae growth and neutral lipid synthesis which is proposed by Surisetty et al. is investigated for the purpose of maximizing the rate of microalgae growth and the amount of neutral lipid. The model has 6 states and 12 parameters and follows the assumption of Droop model which explains the growth as a two-step phenomenon; the uptake of nutrients is first occurred in the cell, and then use of intracellular nutrient to support cell's growth. In this study, optimal experimental design (OED) using D- optimality criterion is performed to compute the system input profile and find optimal parameters of the model. Sensitivity analysis is also performed to determine which parameters have a negligible effect on the model predictions.