In this present study, microalgal phycobiliproteins were isolated and purified via potential biphasic processing technique for pharmaceutical as well as food applications. The algal pre-treatment techniques were studied to enhance the yield of microalgal phycobiliproteins from the biomass. The proposed methods were optimised to obtain the best recovery yield of phycobiliproteins that can be isolated from the biomass. The phycobiliproteins were further purified using liquid biphasic system. The results showed that microalgal phycobiliproteins of high purity and yield was achieved using sonication treatment (20% power, 50% duty cycle and 7 min of irradiation time) with the biphasic system, where the purification fold of 6.17 and recovery yield of 94.89% was achieved. This work will provide insights towards the effective downstream processing of biomolecules from microalgae.
Bioprospecting for biodiesel potential in microalgae primarily involves a few model species of microalgae and rarely on non-model microalgae species. Therefore, the present study determined changes in physiology, oil accumulation, fatty acid composition and biodiesel properties of a non-model microalga Messastrum gracile SE-MC4 in response to 12 continuous days of nitrate-starve (NS) and nitrate-replete (NR) conditions respectively. Under NS, the highest oil content (57.9%) was achieved despite reductions in chlorophyll content, biomass productivity and lipid productivity. However, under both NS and NR, palmitic acid and oleic acid remained as dominant fatty acids thus suggesting high potential of M. gracile for biodiesel feedstock consideration. Biodiesel properties analysis returned high values of cetane number (CN 61.9-64.4) and degree of unsaturation (DU 45.3-57.4) in both treatments. The current findings show the possibility of a non-model microalga to inherit superior ability over model species in oil accumulation for biodiesel development.
Docosahexaenoic acid (DHA, C22:6n-3) plays a vital role in the enhancement of human health, particularly for cognitive, neurological, and visual functions. Marine microalgae, such as members of the genus Aurantiochytrium, are rich in DHA and represent a promising source of omega-3 fatty acids. In this study, levels of glucose, yeast extract, sodium glutamate and sea salt were optimized for enhanced lipid and DHA production by a Malaysian isolate of thraustochytrid, Aurantiochytrium sp. SW1, using response surface methodology (RSM). The optimized medium contained 60 g/L glucose, 2 g/L yeast extract, 24 g/L sodium glutamate and 6 g/L sea salt. This combination produced 17.8 g/L biomass containing 53.9% lipid (9.6 g/L) which contained 44.07% DHA (4.23 g/L). The optimized medium was used in a scale-up run, where a 5 L bench-top bioreactor was employed to verify the applicability of the medium at larger scale. This produced 24.46 g/L biomass containing 38.43% lipid (9.4 g/L), of which 47.87% was DHA (4.5 g/L). The total amount of DHA produced was 25% higher than that produced in the original medium prior to optimization. This result suggests that Aurantiochytrium sp. SW1 could be developed for industrial application as a commercial DHA-producing microorganism.
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.
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.
Green microalga Ankistrodesmus convolutus Corda is a fast growing alga which produces appreciable amount of carotenoids and polyunsaturated fatty acids. To our knowledge, this is the first report on the construction of cDNA library and preliminary analysis of ESTs for this species. The titers of the primary and amplified cDNA libraries were 1.1×10(6) and 6.0×10(9) pfu/ml respectively. The percentage of recombinants was 97% in the primary library and a total of 337 out of 415 original cDNA clones selected randomly contained inserts ranging from 600 to 1,500 bps. A total of 201 individual ESTs with sizes ranging from 390 to 1,038 bps were then analyzed and the BLASTX score revealed that 35.8% of the sequences were classified as strong match, 38.3% as nominal and 25.9% as weak match. Among the ESTs with known putative function, 21.4% of them were found to be related to gene expression, 14.4% ESTs to photosynthesis, 10.9% ESTs to metabolism, 5.5% ESTs to miscellaneous, 2.0% to stress response, and the remaining 45.8% were classified as novel genes. Analysis of ESTs described in this paper can be an effective approach to isolate and characterize new genes from A. convolutus and thus the sequences obtained represented a significant contribution to the extensive database of sequences from green microalgae.
A facile chemical reduction approach is adopted for the synthesis of iron tungstate (FeWO4)/ceria (CeO2)-decorated reduced graphene oxide (rGO) nanocomposite. Surface morphological studies of rGO/FeWO4/CeO2 composite reveal the formation of hierarchical FeWO4 flower-like microstructures on rGO sheets, in which the CeO2 nanoparticles are decorated over the FeWO4 microstructures. The distinct anodic peaks observed for the cyclic voltammograms of studied electrodes under light/dark regimes validate the electroactive proteins present in the microalgae. With the cumulative endeavors of three-dimensional FeWO4 microstructures, phase effect between rGO sheet and FeWO4/CeO2, highly exposed surface area, and light harvesting property of CeO2 nanoparticles, the relevant rGO/FeWO4/CeO2 nanocomposite demonstrates high power and stable biophotovoltaic energy generation compared with those of previous reports. Thus, these findings construct a distinct horizon to tailor a ternary nanocomposite with high electrochemical activity for the construction of cost-efficient and environmentally benign fuel cells.
Marine phycosphere harbors unique cross-kingdom associations with enormous ecological significance in aquatic ecosystems as well as relevance for algal biotechnology industry. During our investigating the microbial composition and bioactivity of marine phycosphere microbiota (PM), a novel lightly yellowish and versatile bacterium designated strain AM1-D1T was isolated from cultivable PM of marine dinoflagellate Alexandrium minutum amtk4 that produces high levels of paralytic shellfish poisoning toxins (PSTs). Strain AM1-D1T demonstrates notable bioflocculanting bioactivity with bacterial exopolysaccharides (EPS), and microalgae growth-promoting (MGP) potential toward its algal host. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain AM1-D1T was affiliated to the members of genus Sulfitobacter within the family Rhodobacteraceae, showing the highest sequence similarity of 97.9% with Sulfitobacter noctilucae NB-68T, and below 97.8% with other type strains. The complete genome of strain AM1-D1T consisted of a circular 3.84-Mb chromosome and five circular plasmids (185, 95, 15, 205 and 348 Kb, respectively) with the G+C content of 64.6%. Low values obtained by phylogenomic calculations on the average nucleotide identity (ANI, 77.2%), average amino acid identity (AAI, 74.7%) and digital DNA-DNA hybridization (dDDH, 18.6%) unequivocally separated strain AM1-D1T from its closest relative. The main polar lipids were identified as phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, one unidentified phospholipid and one unidentified lipid. The predominant fatty acids (> 10%) were C18:1 ω7c, C19:0 cyclo ω8c and C16:0. The respiratory quinone was Q-10. The genome of strain AM1-D1T was predicted to encode series of gene clusters responsible for sulfur oxidation (sox) and utilization of dissolved organic sulfur exometabolites from marine dinoflagellates, taurine (tau) and dimethylsulfoniopropionate (DMSP) (dmd), as well as supplementary vitamin B12 (cob), photosynthesis carotenoids (crt) which are pivotal components during algae-bacteria interactions. Based on the evidences by the polyphasic characterizations, strain AM1-D1T represents a novel species of the genus Sulfitobacter, for which the name Sulfitobacter alexandrii sp. nov. is proposed. The type strain is AM1-D1T (= CCTCC 2017277T = KCTC 62491T).
Microalgae have received much interest as a biofuel feedstock in response to the uprising energy crisis, climate change and depletion of natural sources. Development of microalgal biofuels from microalgae does not satisfy the economic feasibility of overwhelming capital investments and operations. Hence, high-value co-products have been produced through the extraction of a fraction of algae to improve the economics of a microalgae biorefinery. Examples of these high-value products are pigments, proteins, lipids, carbohydrates, vitamins and anti-oxidants, with applications in cosmetics, nutritional and pharmaceuticals industries. To promote the sustainability of this process, an innovative microalgae biorefinery structure is implemented through the production of multiple products in the form of high value products and biofuel. This review presents the current challenges in the extraction of high value products from microalgae and its integration in the biorefinery. The economic potential assessment of microalgae biorefinery was evaluated to highlight the feasibility of the process.
Microalgae technology, if managed properly, has promising roles in solving food-water-energy nexus. The Achilles' heel is, however, to lower the costs associated with cultivation and harvesting. As a favorable technique, application of membrane process is strongly limited by membrane fouling. This study evaluates performance of nylon 6,6 nanofiber membrane (NFM) to a conventional polyvinylidene fluoride phase inverted membrane (PVDF PIM) for filtration of Chlorella vulgaris. Results show that nylon 6,6 NFM is superhydrophilic, has higher size of pore opening (0.22 vs 0.18 μm) and higher surface pore density (23 vs 18 pores/μm2) leading to higher permeance (1018 vs 493 L/m2hbar) and better fouling resistant. Such advantages help to outperform the filterability of PVDF PIM by showing much higher steady-state permeance (286 vs 120 L/m2hbar), with comparable biomass retention. In addition, unlike for PVDF PIM, imposing longer relaxation cycles further enhances the performance of the NFM (i.e., 178 L/m2hbar for 0.5 min and 236 L/m2hbar for 5 min). Overall findings confirm the advantages of nylon 6,6 NFM over the PVDF PIM. Such advantages can help to reduce required membrane area and specific aeration demand by enabling higher flux and lowering aeration rate. Nevertheless, developments of nylon 6,6 NFM material with respect to its intrinsic properties, mechanical strength and operational conditions of the panel can still be explored to enhance its competitiveness as a promising fouling resistant membrane material for microalgae filtration.
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.
Cesium-137 (137Cs) is one of the radioactive substances that was released into the environment as a result of the Fukushima nuclear disaster. Radiocesium exposure is of great concern due to its potential environmental implications. However, research on 137Cs removal using algae is still limited. This is the first report to describe the kinetic properties of 137Cs uptake by Vacuoliviride crystalliferum in the presence and absence of potassium. In this work, we studied the kinetic properties of 137Cs uptake using a freshwater microalga, V. crystalliferum (NIES 2860). We also analyzed the effects of temperature, light, and potassium (K) on the 137Cs uptake. Results showed that V. crystalliferum can remove up to 90% of 157 nM 137Cs within an hour. At 20 °C, the removal increased by up to 96%, compared to less than 10% at 5 °C. However, the removal was inhibited by nearly 90% in the dark compared to the removal in the light, implying that V. crystalliferum cells require energy to accumulate 137Cs. In the inhibition assay, K concentrations ranged from 0 to 500 µM and the inhibitory constant (Ki) for K was determined to be 16.7 µM. While in the uptake assay without potassium (- K), the Michaelis constant (Km) for Cs was 45 nM and increased to 283 nM by the addition of 20 µM potassium (+ K), indicating that V. crystalliferum had a high affinity for 137Cs. In addition, the maximum uptake velocity (Vmax) also increased from 6.75 to 21.10 nmol (mg Chl h)-1, implying the existence of Cs active transport system. In conclusion, V. crystalliferum is capable of removing radioactive 137Cs from the environment and the removal was favorable at both normal temperature and in the light.
In viral respiratory infections, disrupted pathophysiological outcomes have been attributed to hyper-activated and unresolved inflammation responses of the immune system. Integration between available drugs and natural therapeutics have reported benefits in relieving inflammation-related physiological outcomes and microalgae may be a feasible source from which to draw from against future coronavirus-infections. Microalgae represent a large and diverse source of chemically functional compounds such as carotenoids and lipids that possess various bioactivities, including anti-inflammatory properties. Therefore in this paper, some implicated pathways causing inflammation in viral respiratory infections are discussed and juxtaposed along with available research done on several microalgal metabolites. Additionally, the therapeutic properties of some known anti-inflammatory, antioxidant and immunomodulating compounds sourced from microalgae are reported for added clarity.
In the present study, a novel oleaginous Thraustochytrid containing a high content of docosahexaenoic acid (DHA) was isolated from a mangrove ecosystem in Malaysia. The strain identified as an Aurantiochytrium sp. by 18S rRNA sequencing and named KRS101 used various carbon and nitrogen sources, indicating metabolic versatility. Optimal culture conditions, thus maximizing cell growth, and high levels of lipid and DHA production, were attained using glucose (60 g l⁻¹) as carbon source, corn steep solid (10 g l⁻¹) as nitrogen source, and sea salt (15 g l⁻¹). The highest biomass, lipid, and DHA production of KRS101 upon fed-batch fermentation were 50.2 g l⁻¹ (16.7 g l⁻¹ day⁻¹), 21.8 g l⁻¹ (44% DCW), and 8.8 g l⁻¹ (40% TFA), respectively. Similar values were obtained when a cheap substrate like molasses, rather than glucose, was used as the carbon source (DCW of 52.44 g l⁻¹, lipid and DHA levels of 20.2 and 8.83 g l⁻¹, respectively), indicating that production of microbial oils containing high levels of DHA can be produced economically when the novel strain is used.
This work aimed to study the application of liquid biphasic flotation (LBF) for the efficient and rapid recovery of astaxanthin from H. pluvialis microalgae. The performance of LBF for the extraction of astaxanthin was studied comprehensively under different operating conditions, including types and concentrations of food-grade alcohol and salt, volume ratio, addition of neutral salt, flotation period, and mass of dried H. pluvialis biomass powder. The maximum recovery, extraction efficiency and partition coefficient of astaxanthin obtained from the optimum LBF system were 95.11 ± 1.35%, 99.84 ± 0.05% and 385.16 ± 3.87, respectively. A scaled-up LBF system was also performed, demonstrating the feasibility of extracting natural astaxanthin from microalgae at a larger scale. This exploration of LBF system opens a promising avenue to the extraction of astaxanthin at lower cost and shorter processing time.
Microalgae lipids and oils are potential candidates for renewable biofuels and nutritional inventions. Recent studies from our lab have shown that two plant hormones, auxin and jasmonic acid, influence microalgae growth and fatty acid accumulation. Therefore, in this study, a high oil-producing strain Chlorella vulgaris UMT-M1 was selected for hormonal study using gibberellin (GA). Exogenous GA3 was applied to early stationary culture of C. vulgaris UMT-M1. Results showed that GA3 gradually increases the cell density of C. vulgaris to up to 42% on days after treatment (DAT)-8 and also capable of delaying the algal senescence. However, the increment in cell density did not enhance the total oil production albeit transient modification of fatty acid compositions was observed for saturated (SFA) and polyunsaturated (PUFA) fatty acids. This illustrates that GA3 only promotes cell division and growth but not the oil accumulation. In addition, application of GA3 in culture medium was shown to promote transient increment of palmitic (C16:0) and stearic (C18:0) acids from DAT-4 to DAT-6 and these changes are correlated with the expression of β-ketoacyl ACP synthase I (KAS I) gene.
The primary biological treatment method for organic sludge is composting and/or anaerobic digestion, but their product (compost or biogas) is of little economic benefit; therefore, an improved process to produce a high-value product is required to make sludge management more sustainable. Maximizing NH3 gas recovery during composting processes has the potential benefit of producing high-value microalgal biomass. However, the majority of produced ammonia does not evaporate as NH3 gas but retains as NH4+-N in the compost after fermentation. The present study investigates the effects of the timing of Ca(OH)2 dosing (on days 2, 5, and 9), and the Ca(OH)2 dose (1.1-2.6 mmol/batch), on lab-scale thermophilic composting of anaerobic sludge. The effects on NH3 recovery, organic matter degradability, and microbial activity are evaluated. Ca(OH)2 dosing immediately improved the emission of NH3, with yields 50-69% higher than those under control conditions. The timing of the dosing did not influence NH3 recovery or organic matter degradability. Higher Ca(OH)2 doses resulted in higher NH3 recovery, while microbial activity was temporarily and marginally inhibited. The pH of the compost reached 10-11.5 but quickly dropped to 8-8.5 within a day, probably because of neutralization of Ca(OH)2 by the emitted CO2 and release of NH3, which maintained the microbial activity. The present study indicated that Ca(OH)2 dosing would be useful to apply during thermophilic composting for NH3 recovery to cultivate high-value microalgal biomass, which enables this process to obtain a more economic benefit.
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.
Conventional microalgae oil extraction applies physicochemical destruction of dry cell biomass prior to transesterification process to produce fatty acid methyl esters (FAMEs). This report presents a simple and rapid direct transesterification (DT) method for FAMEs production and fatty acid profiling of microalgae using freshly harvested biomass. Results revealed that the FAMEs recovered from Chlorella vulgaris were 50.1 and 68.3 mg with conventional oil-extraction-transesterification (OET) and DT method, respectively. While for Messastrum gracile, the FAMEs recovered, were 49.9 and 76.3 mg, respectively with OET and DT methods. This demonstrated that the DT method increased FAMEs recovery by 36.4% and 53.0% from C. vulgaris and M. gracile, respectively, as compared to OET method. Additionally, the DT method recovered a significantly higher amount of palmitic (C16:0) and stearic (C18:0) acids from both species, which indicated the important role of these fatty acids in the membranes of cells and organelles. The DT method performed very well using a small volume (5 mL) of fresh biomass coupled with a shorter reaction time (~ 15 min), thus making real-time monitoring of FAMEs and fatty acid accumulation in microalgae culture feasible.