Flashing lights affect the photophysiology and expression of carotenoid and lipid synthesis genes in Nannochloropsis gaditana.

Nannochloropsis gaditana is a promising microalga for biotechnology. One of the strategies to stimulate its full potential in metabolite production is exposure to flashing lights. Here, we report how N. gaditana adapts to different flashing light regimes (5, 50, and 500 Hz) by changing its cellular physiology and the relative expression of genes related to critical cellular functions. We analyzed the differential mRNA abundance of genes related to photosynthesis, nitrogen assimilation and biosynthesis of chlorophyll, carotenoids, lipids, fatty acids and starch. Analysis of photosynthetic efficiency and high mRNA abundance of photoprotection genes supported the inference that excess excitation energy provided by light absorbance during photosynthesis was produced under low frequency flashing lights and was dissipated by photopigments via the xanthophyll-cycle. Increased relative expression levels of genes related to the synthesis of carotenoids and chlorophyll confirmed the accumulation of photopigments previously observed at low frequency flashing lights. Higher differential mRNA abundance of genes related to the triacylglycerol biosynthesis were observed at lower frequency flashing lights, possibly triggered by a poor nitrogen assimilation caused by low mRNA abundance of a nitrate reductase gene. This study advances a new understanding of algal physiology and metabolism leading to improved cellular performance and metabolite production.

Drying Microalgae Using an Industrial Solar Dryer: A Biomass Quality Assessment.

Microalgae are considered a promising resource of proteins, lipids, carbohydrates, and other functional biomolecules for food and feed markets. Competitive drying solutions are required to meet future demands for high-quality algal biomass while ensuring proper preservation at reduced costs. Since often used drying methods, such as freeze or spray drying, are energy and time consuming, more sustainable processes remain to be developed. This study tested an indirect and hybrid solar dryer as an alternative to conventional freeze drying of industrially produced Tetraselmis chui and Nannochloropsis oceanica wet paste. The effects of the drying method on biomass quality parameters, including biochemical profiles, functional properties, and microbial safety, were assessed. No significant differences were found between the applied drying technologies for total proteins, carbohydrates, lipids, and fatty acid profiles. On the other hand, some pigments showed significant differences, displaying up to 44.5% higher contents in freeze-dried samples. Minor differences were also registered in the mineral profiles (<10%). Analyses of microbial safety and functional properties of the solar-dried biomass appear adequate for food and feed products. In conclusion, industrial solar drying is a sustainable technology with a high potential to preserve high-quality microalgal biomass for various markets at expected lower costs.

Algae as Food in Europe: An Overview of Species Diversity and Their Application.

Algae have been consumed for millennia in several parts of the world as food, food supplements, and additives, due to their unique organoleptic properties and nutritional and health benefits. Algae are sustainable sources of proteins, minerals, and fiber, with well-balanced essential amino acids, pigments, and fatty acids, among other relevant metabolites for human nutrition. This review covers the historical consumption of algae in Europe, developments in the current European market, challenges when introducing new species to the market, bottlenecks in production technology, consumer acceptance, and legislation. The current algae species that are consumed and commercialized in Europe were investigated, according to their status under the European Union (EU) Novel Food legislation, along with the market perspectives in terms of the current research and development initiatives, while evaluating the interest and potential in the European market. The regular consumption of more than 150 algae species was identified, of which only 20% are approved under the EU Novel Food legislation, which demonstrates that the current legislation is not broad enough and requires an urgent update. Finally, the potential of the European algae market growth was indicated by the analysis of the trends in research, technological advances, and market initiatives to promote algae commercialization and consumption.

Nannochloropsis oceanica harvested using electrocoagulation with alternative electrodes - An innovative approach on potential biomass applications.

Electrocoagulation is a promising technology to harvest microalgal biomass. However, the commonly used aluminum electrodes release undesired salts that decrease biomass value. In this study, alternative iron, zinc, and magnesium electrodes and operational parameters pH, time and current density were studied to harvest Nannochloropsis oceanica. For recovery efficiency and concentration factor the initial pH was most important using iron electrodes, while time and current density were more relevant using zinc and magnesium electrodes. Optimal parameters resulted in biomass recovery efficiencies > 95%, biomass was concentrated 2.8-7.2 times and contained 15.7-29.1% ashes. Elemental analysis revealed metal salts in harvested biomass resulting from electrode corrosion. Finally, ash contents could be reduced by 65% using EDTA as a chelating agent. The electrocoagulation harvested microalgal biomass enriched in essential metals may be a promising bioresource for agricultural growth inducers, or functional ingredients for feed.

Carotenoid biosynthetic gene expression, pigment and n-3 fatty acid contents in carotenoid-rich Tetraselmis striata CTP4 strains under heat stress combined with high light.

In this study, two carotenoid-rich strains of the euryhaline microalga Tetraselmis striata CTP4 were isolated by random mutagenesis combined with selection via fluorescence activated cell sorting and growth on norflurazon. Both strains, ED5 and B11, showed an up to 1.5-fold increase in carotenoid contents as compared with the wildtype, independent of the growth conditions. More specifically, violaxanthin, ß-carotene and lutein contents reached as high as 1.63, 4.20 and 3.81 mg g-1 DW, respectively. Genes coding for phytoene synthase, phytoene desaturase, lycopene-ß-cyclase and ε-ring hydroxylase involved in carotenoid biosynthesis were found to be upregulated in ED5 and B11 cells as compared to the wildtype. Both strains showed higher contents of eicosapentaenoic acid as compared with those of the wildtype, reaching up to 4.41 and 2.88 mg g-1 DW, respectively. Overall, these results highlight the complexity of changes in carotenoid biosynthesis regulation that are required to improve pigment contents in microalgae.

Flashing light emitting diodes (LEDs) induce proteins, polyunsaturated fatty acids and pigments in three microalgae.

As the periodic emission of light pulses by light emitting diodes (LEDs) is known to stimulate growth or induce high value biocompounds in microalgae, this flashing light regime was tested on growth and biochemical composition of the microalgae Nannochloropsis gaditana, Koliella antarctica and Tetraselmis chui. At low flashing light frequencies (e.g., 5 and 50 Hz, Duty cycle = 0.05), a strain-dependent growth inhibition and an accumulation of protein, polyunsaturated fatty acids, chlorophyll or carotenoids (lutein, ß-carotene, violaxanthin and neoxanthin) was observed. In addition, a 4-day application of low-frequency flashing light to concentrated cultures increased productivities of eicosapentaenoic acid (EPA) and specific carotenoids up to three-fold compared to continuous or high frequency flashing light (500 Hz, Duty cycle = 0.05). Therefore, applying low-frequency flashing light as finishing step in industrial production can increase protein, polyunsaturated fatty acids or pigment contents in biomass, leading to high-value algal products.

Temperature-Dependent Lipid Accumulation in the Polar Marine Microalga Chlamydomonas malina RCC2488.

The exploration of cold-adapted microalgae offers a wide range of biotechnological applications that can be used for human, animal, and environmental benefits in colder climates. Previously, when the polar marine microalga Chlamydomonas malina RCC2488 was cultivated under both nitrogen replete and depleted conditions at 8°C, it accumulated lipids and carbohydrates (up to 32 and 49%, respectively), while protein synthesis decreased (up to 15%). We hypothesized that the cultivation temperature had a more significant impact on lipid accumulation than the nitrogen availability in C. malina. Lipid accumulation was tested at three different temperatures, 4, 8, and 15°C, under nitrogen replete and depleted conditions. At 4°C under the nitrogen replete condition C. malina had the maximal biomass productivity (701.6 mg L-1 day-1). At this condition, protein content was higher than lipids and carbohydrates. The lipid fraction was mainly composed of polyunsaturated fatty acids (PUFA) in the polar lipid portion, achieving the highest PUFA productivity (122.5 mg L-1 day-1). At this temperature, under nitrogen deficiency, the accumulation of carbohydrates and neutral lipids was stimulated. At 8 and 15°C, under both nitrogen replete and depleted conditions, the lipid and carbohydrate content were higher than at 4°C, and the nitrogen stress condition did not affect the algal biochemical composition. These results suggest that C. malina is a polar marine microalga with a favorable growth temperature at 4°C and is stressed at temperatures ≥8°C, which directs the metabolism to the synthesis of lipids and carbohydrates. Nevertheless, C. malina RCC2488 is a microalga suitable for PUFA production at low temperatures with biomass productivities comparable with mesophilic strains.

Flashing LEDs for Microalgal Production.

Flashing lights are next-generation tools to mitigate light attenuation and increase the photosynthetic efficiency of microalgal cultivation systems illuminated by light-emitting diodes (LEDs). Optimal flashing light conditions depend on the reaction kinetics and properties of the linear electron transfer chain, energy dissipation, and storage mechanisms of a phototroph. In particular, extremely short and intense light flashes potentially mitigate light attenuation in photobioreactors without impairing photosynthesis. Intelligently controlling flashing light units and selecting electronic components can maximize light emission and energy efficiency. We discuss the biological, physical, and technical properties of flashing lights for algal production. We combine recent findings about photosynthetic pathways, self-shading in photobioreactors, and developments in solid-state technology towards the biotechnological application of LEDs to microalgal production.

Urban wastewater treatment by Tetraselmis sp. CTP4 (Chlorophyta).

The ability of a recent isolate, Tetraselmis sp. CTP4, for nutrient removal from sewage effluents before and after the nitrification process under batch and continuous cultivation was studied. Biomass productivities in both wastewaters were similar under continuous conditions (0.343±0.053gL-1d-1) and nutrient uptake rates were maximal 31.4±0.4mgNL-1d-1 and 6.66±1.57mgP-PO43-L-1d-1 in WW before nitrification when cultivated in batch. Among batch treatments, cellular protein, carbohydrate and lipid levels shifted with aging cultures from 71.7±6.3 to 29.2±1.2%, 17.4±7.2 to 57.2±3.9% and 10.9±1.7 to 13.7±4.7%, respectively. In contrast, CTP4 cultivated continuously in Algal medium (control) showed lower biomass productivities (0.282gVSSL-1d-1) although improved lipid content (up to 20% lipids) in batch cultivation. Overall, Tetraselmis sp. CTP4 is promising for WW treatment as a replacement of the costly nitrification process, fixating more nutrients and providing a protein and carbohydrate-rich biomass as by-product.

Isolation of a euryhaline microalgal strain, Tetraselmis sp. CTP4, as a robust feedstock for biodiesel production.

Bioprospecting for novel microalgal strains is key to improving the feasibility of microalgae-derived biodiesel production. Tetraselmis sp. CTP4 (Chlorophyta, Chlorodendrophyceae) was isolated using fluorescence activated cell sorting (FACS) in order to screen novel lipid-rich microalgae. CTP4 is a robust, euryhaline strain able to grow in seawater growth medium as well as in non-sterile urban wastewater. Because of its large cell size (9-22 µm), CTP4 settles down after a six-hour sedimentation step. This leads to a medium removal efficiency of 80%, allowing a significant decrease of biomass dewatering costs. Using a two-stage system, a 3-fold increase in lipid content (up to 33% of DW) and a 2-fold enhancement in lipid productivity (up to 52.1 mg L-1 d-1) were observed upon exposure to nutrient depletion for 7 days. The biodiesel synthesized from the lipids of CTP4 contained high levels of oleic acid (25.67% of total fatty acids content) and minor amounts of polyunsaturated fatty acids with ≥4 double bonds (<1%). As a result, this biofuel complies with most of the European (EN14214) and American (ASTM D6751) specifications, which commonly used microalgal feedstocks are usually unable to meet. In conclusion, Tetraselmis sp. CTP4 displays promising features as feedstock with lower downstream processing costs for biomass dewatering and biodiesel refining.

Light emitting diodes (LEDs) applied to microalgal production.

Light-emitting diodes (LEDs) will become one of the world's most important light sources and their integration in microalgal production systems (photobioreactors) needs to be considered. LEDs can improve the quality and quantity of microalgal biomass when applied during specific growth phases. However, microalgae need a balanced mix of wavelengths for normal growth, and respond to light differently according to the pigments acquired or lost during their evolutionary history. This review highlights recently published results on the effect of LEDs on microalgal physiology and biochemistry and how this knowledge can be applied in selecting different LEDs with specific technical properties for regulating biomass production by microalgae belonging to diverse taxonomic groups.