This study aimed to assess the inhibitory activities of methanol extracts from the microalgae Ankistrodesmus convolutus, Synechococcus elongatus, and Spirulina platensis against Epstein-Barr virus (EBV) in three Burkitt's lymphoma (BL) cell lines, namely Akata, B95-8, and P3HR-1. The antiviral activity was assessed by quantifying the cell-free EBV DNA using real-time polymerase chain reaction (PCR) technique. The methanol extracts from Ankistrodesmus convolutus and Synechococcus elongatus displayed low cytotoxicity and potent effect in reducing cell-free EBV DNA (EC(50)<0.01 µg/ml) with a high therapeutic index (>28000). After fractionation by column chromatography, the fraction from Synechococcus elongatus (SEF1) reduced the cell-free EBV DNA most effectively (EC(50)=2.9 µg/ml, therapeutic index>69). Upon further fractionation by high performance liquid chromatography (HPLC), the sub-fraction SEF1'a was most active in reducing the cell-free EBV DNA (EC(50)=1.38 µg/ml, therapeutic index>14.5). This study suggests that microalgae could be a potential source of antiviral compounds that can be used against EBV.
Microalgae lipids and oils are potential candidates for renewable biodiesel. Many microalgae species accumulate a substantial amount of lipids and oils under environmental stresses. However, low growth rate under these adverse conditions account for the decrease in overall biomass productivity which directly influence the oil yield. This study was undertaken to investigate the effect of exogenously added auxin (indole-3-acetic acid; IAA) on the oil content, fatty acid compositions, and the expression of fatty acid biosynthetic genes in Chlorella vulgaris (UMT-M1). Auxin has been shown to regulate growth and metabolite production of several microalgae. Results showed that oil accumulation was highest on days after treatment (DAT)-2 with enriched levels of palmitic (C16:0) and stearic (C18:0) acids, while the linoleic (C18:2) and α-linolenic (C18:3n3) acids levels were markedly reduced by IAA. The elevated levels of saturated fatty acids (C16:0 and C18:0) were consistent with high expression of the β-ketoacyl ACP synthase I (KAS I) gene, while low expression of omega-6 fatty acid desaturase (ω-6 FAD) gene was consistent with low production of C18:2. However, the increment of stearoyl-ACP desaturase (SAD) gene expression upon IAA induction did not coincide with oleic acid (C18:1) production. The expression of omega-3 fatty acid desaturase (ω-3 FAD) gene showed a positive correlation with the synthesis of PUFA and C18:3n3.
Marine microalgae have been prominently featured in cancer research. Here, we examined cytotoxic effect and apoptosis mechanism of crude ethanol extracts of an indigenous microalga, Chaetoceros calcitrans (UPMAAHU10) on human breast cell lines. MCF-7 was more sensitive than MCF-10A with IC50 value of 3.00 ± 0.65, whilst the IC50 value of Tamoxifen against MCF-7 was 12.00 ± 0.52 μg/mL after 24 hour incubation. Based on Annexin V/Propidium iodide and cell cycle flow cytometry analysis, it was found that inhibition of cell growth by EEC on MCF-7 cells was through the induction of apoptosis without cell cycle arrest. The apoptotic cells at subG0/G1 phase in treated MCF-7 cells at 48 and 72 hours showed 34 and 16 folds increased compared to extract treated MCF-10A cells which showed only 6 and 7 folds increased at the same time points, respectively. Based on GeXP study, EEC induced apoptosis on MCF-7 cells via modulation of CDK2, MDM2, p21Cip1, Cyclin A2, Bax and Bcl-2. The EEC treated MCF-7 cells also showed an increase in Bax/Bcl-2 ratio that in turn activated the caspase-dependent pathways by activating caspase 7. Thus, marine microalga, Chaetoceros calcitrans may be considered a good candidate to be developed as a new anti-breast cancer drug.
Global warming has become a serious issue nowadays as the trend of CO2 emission is increasing by years. In Malaysia, the electricity and energy sector contributed a significant amount to the nation's CO2 emission due to fossil fuel use. Many research works have been carried out to mitigate this issue, including carbon capture and utilization (CCUS) technology and biological carbon fixation by microalgae. This study makes a preliminary effort to screen native microalgae species in the Malaysian coal-fired power plant's surrounding towards carbon fixation ability. Three dominant species, including Nannochloropsis sp., Tetraselmis sp., and Isochrysis sp. were identified and tested in the laboratory under ambient and pure CO2 condition to assess their growth and CO2 fixation ability. The results indicate Isochrysis sp. as the superior carbon fixer against other species. In continuation, the optimization study using Response Surface Methodology (RSM) was carried out to optimize the operating conditions of Isochrysis sp. using a customized lab-scale photobioreactor under simulated flue gas exposure. This species was further acclimatized and tested under actual flue gas generated by the power plant. Isochrysis sp. had shown its capability as a carbon fixer with CO2 fixation rate of 0.35 gCO2/L day under actual coal-fired flue gas exposure after cycles of acclimatization phase. This work is the first to demonstrate indigenous microalgae species' ability as a carbon fixer under Malaysian coal-fired flue gas exposure. Thus, the findings shall be useful in exploring the microalgae potential as a biological agent for carbon emission mitigation from power plants more sustainably.
Microalgae are the major photosynthesizers on earth and produce important pigments that include chlorophyll a, b and c, β-carotene, astaxanthin, xanthophylls, and phycobiliproteins. Presently, synthetic colorants are used in food, cosmetic, nutraceutical, and pharmaceutical industries. However, due to problems associated with the harmful effects of synthetic colorants, exploitation of microalgal pigments as a source of natural colors becomes an attractive option. There are various factors such as nutrient availability, salinity, pH, temperature, light wavelength, and light intensity that affect pigment production in microalgae. This paper reviews the availability and characteristics of microalgal pigments, factors affecting pigment production, and the application of pigments produced from microalgae. The potential of microalgal pigments as a source of natural colors is enormous as an alternative to synthetic coloring agents, which has limited applications due to regulatory practice for health reasons.
The types of microalgae strains and the method used in lipid extraction have become crucial factors which influence the productivity of crude oil. In this paper, Nannochloropsis sp. and Tetraselmis sp. were chosen as the strains and four different methods were used to extract the lipids: Hara and Radin, Folch, Chen and Bligh and Dyer. These methods were performed by using conventional heating and microwave irradiation methods. Results revealed that highest lipid yield from the different species was obtained using different extraction methods; both under microwave irradiation. The lipid yield for Tetraselmis sp. and Nannochloropsis sp. was highest when Hara and Radin (8.19%), and Folch (8.47%) methods were used respectively under microwave irradiation. The lipids extracted were then transesterified to biodiesel and the quality of the biodiesel was analyzed using the gas chromatography.
Utilization of macroalgae biomass for bioethanol production appears as an alternative source to lignocellulosic materials. In this study, for the first time, Amberlyst (TM)-15 was explored as a potential catalyst to hydrolyze carbohydrates from Eucheuma cottonii extract to simple reducing sugar prior to fermentation process. Several important hydrolysis parameters were studied for process optimization including catalyst loading (2-5%, w/v), reaction temperature (110-130°C), reaction time (0-2.5 h) and biomass loading (5.5-15.5%, w/v). Optimum sugar yield of 39.7% was attained based on the following optimum conditions: reaction temperature at 120°C, catalyst loading of 4% (w/v), 12.5% (w/v) of biomass concentration and reaction time of 1.5h. Fermentation of the hydrolysate using Saccharomyces cerevisiae produced 0.33 g/g of bioethanol yield with an efficiency of 65%. The strategy of combining heterogeneous-catalyzed hydrolysis and fermentation with S. cerevisiae could be a feasible strategy to produce bioethanol from macroalgae biomass.
The wet biomass microalgae of Nannochloropsis sp. was converted to biodiesel using direct transesterification (DT) by microwave technique and ionic liquid (IL) as the green solvent. Three different ionic liquids; 1-butyl-3-metyhlimidazolium chloride ([BMIM][Cl], 1-ethyl-3-methylimmidazolium methyl sulphate [EMIM][MeSO4] and 1-butyl-3-methylimidazolium trifluoromethane sulfonate [BMIM][CF3SO3]) and organic solvents (hexane and methanol) were used as co-solvents under microwave irradiation and their performances in terms of percentage disruption, cell walls ruptured and biodiesel yields were compared at different reaction times (5, 10 and 15 min). [EMIM][MeSO4] showed highest percentage cell disruption (99.73%) and biodiesel yield (36.79% per dried biomass) after 15 min of simultaneous reaction. The results demonstrated that simultaneous extraction-transesterification using ILs and microwave irradiation is a potential alternative method for biodiesel production.
Fish meal is currently the major protein source for commercial aquaculture feed. Due to its unstable supply and increasing price, fish meal is becoming more expensive and its availability is expected to face significant challenges in the near future. Therefore, feasible alternatives to fish meal are urgently required. Microalgae have been recognized as the most promising candidates to replace fish meal because the protein composition of microalgae is similar to fish meal and the supply of microalgae-based proteins is sustainable. In this study, an indigenous microalga (Chlorella vulgaris FSP-E) with high protein content was selected, and its feasibility as an aquaculture protein source was explored. An innovative photobioreactor (PBR) utilizing cold cathode fluorescent lamps as an internal light source was designed to cultivate the FSP-E strain for protein production. This PBR could achieve a maximum biomass and protein productivity of 699 and 365 mg/L/day, respectively, under an optimum urea and iron concentration of 12.4 mM and 90 μM, respectively. In addition, amino acid analysis of the microalgal protein showed that up to 70% of the proteins in this microalgal strain consist of indispensable amino acids. Thus, C. vulgaris FSP-E appears to be a viable alternative protein source for the aquaculture industry.
Cell immobilization on the magnetic nanoparticles (MNPs) and magnetic harvesting is a novel approach for microalgal cells separation. To date, the effect of these nanoparticles on microalgal cells was only studied over a short period of time. More studies are hence needed for a better understanding of the magnetic harvesting proposes or environmental concerns relating to long-term exposure to nanoparticles. In this study, the impact of various concentrations of MNPs on the microalgal cells growth and their metabolic status was investigated over 12 days. More than 60% reduction in mitochondrial activity and pigments (chlorophyll a, chlorophyll b, and carotenoids) content occurred during the first 6 days of exposure to ≥50 µg/mL nanoparticles. However, more than 50% growth inhibitory effect was seen at concentrations higher than 400 µg/mL. Exposure to MNPs gradually induced cellular adaptation and after about 6 days of exposure to stress generating concentrations (˂400 µg/mL) of IONs, microalgae could overcome the imposed damages. This work provides a better understanding regarding the environmental impact of MNPs and appropriate concentrations of these particles for future algal cells magnetic immobilization and harvesting.
The richness of high-value bio-compounds derived from microalgae has made microalgae a promising and sustainable source of useful product. The present work starts with a review on the usage of open pond and photobioreactor in culturing various microalgae strains, followed by an in-depth evaluation on the common harvesting techniques used to collect microalgae from culture medium. The harvesting methods discussed include filtration, centrifugation, flocculation, and flotation. Additionally, the advanced extraction technologies using ionic liquids as extractive solvents applied to extract high-value bio-compounds such as lipids, carbohydrates, proteins, and other bioactive compounds from microalgae biomass are summarized and discussed. However, more work needs to be done to fully utilize the potential of microalgae biomass for the application in large-scale production of biofuels, food additives, and nutritive supplements.
Microalgae are known as a rich source of bioactive compounds which exhibit different biological activities. Increased demand for sustainable biomass for production of important bioactive components with various potential especially therapeutic applications has resulted in noticeable interest in algae. Utilisation of microalgae in multiple scopes has been growing in various industries ranging from harnessing renewable energy to exploitation of high-value products. The focuses of this review are on production and the use of value-added components obtained from microalgae with current and potential application in the pharmaceutical, nutraceutical, cosmeceutical, energy and agri-food industries, as well as for bioremediation. Moreover, this work discusses the advantage, potential new beneficial strains, applications, limitations, research gaps and future prospect of microalgae in industry.
Natural antioxidants from sustainable sources are favoured to accommodate worldwide antioxidant demand. In addition to bioprospecting for natural and sustainable antioxidant sources, this study aimed to investigate the relationship between the bioactives (i.e. carotenoid and phenolic acids) and the antioxidant capacities in fucoxanthin-producing algae. Total carotenoid, phenolic acid, fucoxanthin contents and fatty acid profile of six species of algae (five microalgae and one macroalga) were quantified followed by bioactivity evaluation using four antioxidant assays. Chaetoceros calcitrans and Isochrysis galbana displayed the highest antioxidant activity, followed by Odontella sinensis and Skeletonema costatum which showed moderate bioactivities. Phaeodactylum tricornutum and Saccharina japonica exhibited the least antioxidant activities amongst the algae species examined. Pearson correlation and multiple linear regression showed that both carotenoids and phenolic acids were significantly correlated (p<0.05) with the antioxidant activities, indicating the influence of these bioactives on the algal antioxidant capacities.
One of the essential fatty acids with therapeutic impacts on human health is known to be omega-3 polyunsaturated fatty acids (PUFA). More lately, ionic liquids (ILs) have received significant attention among scientists in overcoming the disadvantages of traditional solvents in biomass lipid extraction. However, the large pool of cations and anions possibly accessible will lead to a growing number of innovatively synthesized ILs. Nevertheless, the exhaustive measurement of all these systems is economically impractical. The conductive screening model for real solvents (COSMO-RS) is considered a precious approach with the availability of a few models to predict the characteristics of ILs. This work introduces the estimate of capacity values at infinite dilution for a range of ILs using COSMO-RS software as part of solid-liquid extraction. This favorable outcome presented that the capacity values of the IL molecules are extremely dependent on both anions and cations. Among the 352 combinations of cation/anion tested, short alkyl chain cations coupled with inorganic anions were found to be most efficient and therefore superior in the extraction method. Sulphate-, chloride-, and bromide-based ILs were found to have higher extraction capacities in contrast with the remainders, while propanoate revealed an extraordinary capacity when combined with ethyl-based cations. Eventually, the predicted results from COSMO-RS were validated through the experimentally calculated extraction yield of alpha-linolenic acid (ALA) compound from Nannochloropsis sp. microalgae. Three selected ILs namely [EMIM][Cl], [TMAm][Cl], and [EMPyrro][Br] were selected from COSMO-RS for empirical extraction purpose and the validation results pinpointed the good prediction capability of COSMO-RS.
The continuous growth in global population and the ongoing development of countries such as China and India have contributed to a rapid increase in worldwide energy demand. Fossil fuels such as oil and gas are finite resources, and their current rate of consumption cannot be sustained. This, coupled with fossil fuels' role as pollutants and their contribution to global warming, has led to increased interest in alternative sources of energy production. Bioethanol, presently produced from energy crops, is one such promising alternative future energy source and much research is underway in optimizing its production. The economic and temporal constraints that crop feedstocks pose are the main downfalls in terms of the commercial viability of bioethanol production. As an alternative to crop feedstocks, significant research efforts have been put into utilizing algal biomass as a feedstock for bioethanol production. Whilst the overall process can vary, the conversion of biomass to bioethanol usually contains the following steps: (i) pretreatment of feedstock; (ii) hydrolysis; and (iii) fermentation of bioethanol. This paper reviews different technologies utilized in the pretreatment and fermentation steps, and critically assesses their applicability to bioethanol production from algal biomass. Two different established fermentation routes, single-stage fermentation and two-stage gasification/fermentation processes, are discussed. The viability of algal biomass as an alternative feedstock has been assessed adequately, and further research optimisation must be guided toward the development of cost-effective scalable methods to produce high bioethanol yield under optimum economy.
Culturing of microalgae as an alternative feedstock for biofuel production has received a lot of attention in recent years due to their fast growth rate and ability to accumulate high quantity of lipid and carbohydrate inside their cells for biodiesel and bioethanol production, respectively. In addition, this superior feedstock offers several environmental benefits, such as effective land utilization, CO(2) sequestration, self-purification if coupled with wastewater treatment and does not trigger food versus fuel feud. Despite having all these 'theoretical' advantages, review on problems and issues related to energy balance in microalgae biofuel are not clearly addressed until now. Base on the maturity of current technology, the true potential of microalgae biofuel towards energy security and its feasibility for commercialization are still questionable. Thus, this review is aimed to depict the practical problems that are facing the microalgae biofuel industry, covering upstream to downstream activities by accessing the latest research reports and critical data analysis. Apart from that, several interlink solutions to the problems will be suggested with the purpose to bring current microalgae biofuel research into a new dimension and consequently, to revolutionize the entire microalgae biofuel industry towards long-term sustainability.
An aqueous two-phase system (ATPS) with ionic liquids (ILs) was used for the isolate of C-phycocyanin (CPC) from Spirulina platensis microalga. Various imidazolium ILs and potassium salts were studied. The effect of ILs-ATPS on the extraction efficiency of CPC was also studied. The experimental parameters like pH, loading volume, algae concentration, temperature, and alkyl chain length of IL were well-covered in this report. The experimental results showed that the extraction efficiency, the partition coefficient, and the separation factor for CPC were 99%, 36.6, and 5.8, respectively, for an optimal pH value of 7 and a temperature of 308 K. The order of extraction efficiency for CPC using IL-ATPS was: 1-octyl-3-methylimidazolium bromide (C8MIM-Br) > 1-hexyl-3-methylimidazolium bromide (C6MIM-Br) > 1-butyl-3-methylimidazolium bromide (C4MIM-Br). The isolation process followed the pseudo second-order kinetic model and the thermodynamic results were obviously spontaneous.
Lutein available in the current market is derived from marigold petals. However, extensive studies showed that microalgae are rich in lutein content and potentially exploitable for its dietary and other industrial applications. In this study, microwave assisted binary phase solvent extraction method (MABS) was the novel protocol being developed and optimized to achieve maximum lutein recovery from microalgae Scenedesmus sp. biomass. Results showed that 60% potassium hydroxide solution with acetone in the ratio of 0.1 (ml/ml) was the ideal binary phase solvent composition. Empirical model developed using response surface methodology revealed highest lutein content can be recovered through MABS extraction method at 55 °C treatment temperature, 36 min in extraction time, 0.7 (mg/ml) for biomass to solvent ratio, 250 Watt microwave power and 250 rpm stirring speed. This optimized novel protocol had increased the amount of lutein recovered by 130% and shorten the overall extraction time by 3-folds.
The kinetics of lipid extraction utilizing microwave-assisted extraction (MAE) from Nannochloropsis sp. microalgae were studied using a low cost and green solvent, namely brine (NaCl) solution. The kinetic modelling of the lipid extraction was performed to evaluate the mechanism of the lipid mass transfer using different extraction models, including Fick's Law, First and Second-order Rate Law and the Patricelli mathematical model. The Patricelli mathematical model described the kinetics of lipid extraction well, with the highest average values of determination coefficient (R2 ≥ 0.952) and the lowest average values of mean relative percentage deviation (MRPD ≤ 8.666%). The lipid analysis indicated a positive influence of the microwave temperature and time on the quantity and quality of extracted lipids. SEM analysis of spent microalgae clearly shows an increase in the distorted cell with increase microwave temperature and time, which could be directly correlated to the mechanism of the MAE-brine technique.
Microalgal biomass produced from the phycoremediation of wastewater represents an important protein source, lipids, and natural antioxidants and bioproducts. Therefore, the microalgal biomass and their derived compounds are used in animal and aquaculture feed as well as human nutrition and health products. Many microalgal species have shown promising potential for many bioproducts. However, significant processes to find the optimum quality and quantity of microalgal biomass are still required especially when it is used as a replacement for aquaculture feed. The limitations lie in the selection of microalgal species and their production. The present review discusses the potential generation of bioproducts from microalgal biomass resulting from the phycoremediation of wet market wastewater. The consortium approach in wastewater treatment and the comparison between biomass production and available common feeds for aquaculture were reviewed.