De novo transcriptome and lipidome analysis of Desmodesmus abundans under model flue gas reveals adaptive changes after ten years of acclimation to high CO2.

Microalgae's ability to mitigate flue gas is an attractive technology that can valorize gas components through biomass conversion. However, tolerance and growth must be ideal; therefore, acclimation strategies are suggested. Here, we compared the transcriptome and lipidome of Desmodesmus abundans strains acclimated to high CO2 (HCA) and low CO2 (LCA) under continuous supply of model flue gas (MFG) and incomplete culture medium (BG11-N-S). Initial growth and nitrogen consumption from MFG were superior in strain HCA, reaching maximum productivity a day before strain LCA. However, similar productivities were attained at the end of the run, probably because maximum photobioreactor capacity was reached. RNA-seq analysis during exponential growth resulted in 16,435 up-regulated and 4,219 down-regulated contigs in strain HCA compared to LCA. Most differentially expressed genes (DEGs) were related to nucleotides, amino acids, C fixation, central carbon metabolism, and proton pumps. In all pathways, a higher number of up-regulated contigs with a greater magnitude of change were observed in strain HCA. Also, cellular component GO terms of chloroplast and photosystems, N transporters, and secondary metabolic pathways of interest, such as starch and triacylglycerols (TG), exhibited this pattern. RT-qPCR confirmed N transporters expression. Lipidome analysis showed increased glycerophospholipids in strain HCA, while LCA exhibited glycerolipids. Cell structure and biomass composition also revealed strains differences. HCA possessed a thicker cell wall and presented a higher content of pigments, while LCA accumulated starch and lipids, validating transcriptome and lipidome data. Overall, results showed significant differences between strains, where characteristic features of adaptation and tolerance to high CO2 might be related to the capacity to maintain a higher flux of internal C, regulate intracellular acidification, active N transporters, and synthesis of essential macromolecules for photosynthetic growth.

A comprehensive review on the heterotrophic production of bioactive compounds by microalgae.

Bioactive compounds derived from microalgae have garnered considerable attention as valuable resources for drugs, functional foods, and cosmetics. Among these compounds, photosynthetic pigments and polyunsaturated fatty acids (PUFAs) have gained increasing interest due to their numerous beneficial properties, including anti-oxidant, anti-viral, anti-bacterial, anti-fungal, anti-inflammatory, and anti-tumor effects. Several microalgae species have been identified as rich sources of bioactive compounds, including the Chlorophyceae Dunaliella and Haematococcus, the Bacillariophyta Phaeodactylum and Nitzschia, and the dinoflagellate Crypthecodinium cohnii. However, most of the reported microalgae species primarily grow through autotrophic mechanisms, resulting in low yields and high production costs of bioactive compounds. Consequently, the utilization of heterotrophic microalgae, such as Chromochloris zofingiensis and Nitzschia laevis, has shown significant advantages in the production of astaxanthin and eicosapentaenoic acid (EPA), respectively. These heterotrophic microalgae exhibit superior capabilities in synthesizing target compounds. This comprehensive review provides a thorough examination of the heterotrophic production of bioactive compounds by microalgae. It covers key aspects, including the metabolic pathways involved, the impact of cultivation conditions, and the practical applications of these compounds. The review discusses how heterotrophic cultivation strategies can be optimized to enhance bioactive compound yields, shedding light on the potential of microalgae as a valuable resource for high-value product development.

Simultaneous exposure to nanoplastics and cadmium mitigates microalgae cellular toxicity: Insights from molecular simulation and metabolomics.

In the severe pollution area of nanoplastics (NPs) and cadmium ions (Cd2+), the joint effects of their high environmental concentrations on primary producers may differ from those of low environmental doses. Thus, we investigated the physiological changes, cell morphology, molecular dynamic simulation, phenotypic interactions, and metabolomics responses of C. pyrenoidosa to high environmental concentrations of NPs and Cd2+ after 12-d acclimation. After 12-d cultivation, mono-NPs and mono-Cd2+ reduced cell density and triggered antioxidant enzymes, extracellular polymeric substances (EPS) production, and cell aggregation to defend their unfavorable effects. Based on the molecular dynamic simulation, the chlorine atoms of the NPs and Cd2+ had charge attraction with the nitrogen and phosphorus atoms in the choline and phosphate groups in the cell membrane, thereby NPs and Cd2+ could adsorb on the cells to destroy them. In the joint exposure, NPs dominated the variations of ultrastructure and metabolomics and alleviated the toxicity of NPs and Cd2+. Due to its high environmental concentration, more NPs could compete with the microalgae for Cd2+ and thicken cell walls, diminishing the Cd2+ content and antioxidant enzymes of microalgae. NPs addition also decreased the EPS content, while the bound EPS with -CN bond was kept to detoxicate Cd2+. Metabolomics results showed that the NPs downregulated nucleotide, arachidonic acid, and tryptophan metabolisms, while the Cd2+ showed an opposite trend. Compared with their respective exposures, metabolomics results found the changes in metabolic molecules, suggesting the NPs_Cd2+ toxicity was mitigated by balancing nucleotide, arachidonic acid, tryptophan, and arginine and proline metabolisms. Consequently, this study provided new insights that simultaneous exposure to high environmental concentrations of NPs and Cd2+ mitigated microalgae cellular toxicity, which may change their fates and biogeochemical cycles in aquatic systems.

Intensive Microalgal Cultivation and Tertiary Phosphorus Recovery from Wastewaters via the EcoRecover Process.

Mixed community microalgal wastewater treatment technologies have the potential to advance the limits of technology for biological nutrient recovery while producing a renewable carbon feedstock, but a deeper understanding of their performance is required for system optimization and control. In this study, we characterized the performance of a 568 m3·day-1 Clearas EcoRecover system for tertiary phosphorus removal (and recovery as biomass) at an operating water resource recovery facility (WRRF). The process consists of a (dark) mix tank, photobioreactors (PBRs), and a membrane tank with ultrafiltration membranes for the separation of hydraulic and solids residence times. Through continuous online monitoring, long-term on-site monitoring, and on-site batch experiments, we demonstrate (i) the importance of carbohydrate storage in PBRs to support phosphorus uptake under dark conditions in the mix tank and (ii) the potential for polyphosphate accumulation in the mixed algal communities. Over a 3-month winter period with limited outside influences (e.g., no major upstream process changes), the effluent total phosphorus (TP) concentration was 0.03 ± 0.03 mg-P·L-1 (0.01 ± 0.02 mg-P·L-1 orthophosphate). Core microbial community taxa included Chlorella spp., Scenedesmus spp., and Monoraphidium spp., and key indicators of stable performance included near-neutral pH, sufficient alkalinity, and a diel rhythm in dissolved oxygen.

Choline Dehydrogenase Contributes to Salt Tolerance in Dunaliella through Betaine Synthesis.

In Dunaliella tertiolecta, a microalga renowned for its extraordinary tolerance to high salinity levels up to 4.5 M NaCl, the mechanisms underlying its stress response have largely remained a mystery. In a groundbreaking discovery, this study identifies a choline dehydrogenase enzyme, termed DtCHDH, capable of converting choline to betaine aldehyde. Remarkably, this is the first identification of such an enzyme not just in D. tertiolecta but across the entire Chlorophyta. A 3D model of DtCHDH was constructed, and molecular docking with choline was performed, revealing a potential binding site for the substrate. The enzyme was heterologously expressed in E. coli Rosetta (DE3) and subsequently purified, achieving enzyme activity of 672.2 U/mg. To elucidate the role of DtCHDH in the salt tolerance of D. tertiolecta, RNAi was employed to knock down DtCHDH gene expression. The results indicated that the Ri-12 strain exhibited compromised growth under both high and low salt conditions, along with consistent levels of DtCHDH gene expression and betaine content. Additionally, fatty acid analysis indicated that DtCHDH might also be a FAPs enzyme, catalyzing reactions with decarboxylase activity. This study not only illuminates the role of choline metabolism in D. tertiolecta's adaptation to high salinity but also identifies a novel target for enhancing the NaCl tolerance of microalgae in biotechnological applications.

Are native microalgae consortia able to remove microplastics from wastewater effluents?

Wastewater Treatment Plants (WWTPs) are potential sources of microplastics (MPs) in the aquatic environment. This study aimed to investigate the potential of wastewater-native microalgae consortia to remove MPs from the effluent of two different types of WWTPs as a dual-purpose solution for MPs mitigation and biomass production. For that purpose, the occurrence of MPs from two types of WWTP effluents was analysed over one year. MPs were characterized in terms of morphology (microbead, foam, granule, irregular, filament and film), colour and size. The wastewater characterisation was followed by the removal of MP loads, using native microalgae consortia, pre-adapted to the wastewater effluent. Microalgae consortia evolved naturally through four mitigation assays, adapted to seasonal conditions, such as temperature, photoperiod, and wastewater composition. MPs were present in all the effluent samples, ranging from 52 to 233 MP L-1. The characterisation of MPs indicated a predominance of white and transparent particles, with irregular and filament shapes, mainly under 500 µm in size. The µFTIR analysis revealed that 43% of the selected particles were plastic, with a prevalence of polypropylene (PP) (34%) and polyethylene terephthalate (PET) (30 %). In the mitigation experiments, substantial biomass production was achieved (maximum of 2.6 g L-1 (d.w.)), with successful removal of MPs, ranging from 31 ± 25% to 82 ± 13%. These results show that microalgae growth in wastewater effluents efficiently promotes the removal of MPs, reducing this source of contamination in the aquatic environment, while generating valuable biomass. Additionally, the strategy employed, requires minimal control of culture conditions, simplifying the integration of these systems in real-world WWTP facilities for improved wastewater management.

Comparative dynamic optimization study of batch hydrothermal liquefaction of two microalgal strains for economic bio-oil production.

This work presents dynamic optimization strategies of batch hydrothermal liquefaction of two microalgal species, Aurantiochytrium sp. KRS101 and Nannochloropsis sp. to optimize the reactor temperature profiles. Three dynamic optimization problems are solved to maximize the endpoint biocrude yield, minimize the final time, and minimize the reactor thermal energy. The biocrude maximization and time minimization problems demonstrated 11% and 6.18% increment in the optimal biocrude yields and reduction of 78.2% and 61.66% in batch times compared to the base cases for the microalgae with higher lipid and protein fractions, respectively. The energy minimization problem revealed a significant reduction in the reactor thermal energies to generate the targeted biocrude yields compared to the biocrude maximization. Therefore, the identified optimal temperature trajectories outperformed the conventional fixed temperature profiles and could improve the overall economics of the batch bio-oil production from the algal-based biorefineries by significantly enhancing the reactor performance.

Lipids productivity of cyanobacterium Anabaena vaginicola in an internally illuminated photobioreactor using LED bar lights.

Concerns over environmental issues exists and desire to decrease of their extent, have directed efforts toward green energy production. Growth behavior of Anabaena vaginicola was determined in a photobioreator which illuminated internally (IIPBR) using LED bar light. Excessive heat generated in the IIPBR was taken care of by applying a novel air-cooled system. Further note in experimentation was to find favorable cultivation conditions in the IIPBR for A. vaginicola growth and its lipids production capacity. The following results are expressed: 80 µmol photons m-2 s-1 as light intensity, 0.5 g/l as NaNO3, and 120 ml/min as CO2 amount being expressed in terms of aeration rate. The findings were interpreted in terms of a two-component system where the genes encoded to the relevant proteins are present in cyanobacteria and their expressiveness depends on environmental stress. By determining growth rate constant as 0.11 d-1, the productivity in terms of biomass formation was calculated as 202.6 mg L-1 d-1. While rate of lipids production by the test cyanobacterium is 15.65 mg L-1 d-1. Based on total energy used for IIPBR performance, biomass productivity per unit power input equals to 0.74 g W-1 d-1 and this is in favorable position compared with other photobioreactors.

Synergistic effect of nitrate exposure and heatwaves on the growth, and metabolic activity of microalgae, Chlamydomonas reinhardtii, and Pseudokirchneriella subcapitata.

Aquatic biota are threatened by climate warming as well as other anthropogenic stressors such as eutrophication by phosphates and nitrate. However, it remains unclear how nitrate exposure can alter the resilience of microalgae to climate warming, particularly heatwaves. To get a better understanding of these processes, we investigated the effect of elevated temperature and nitrate pollution on growth, metabolites (sugar and protein), oxidative damage (lipid peroxidation), and antioxidant accumulation (polyphenols, proline) in Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata. The experiment involved a 3 × 3 factorial design, where microalgae were exposed to one of three nitrate levels (5, 50, or 200 mg L-1 NO3-l) at 20 °C for 2 weeks. Subsequently, two heatwave scenarios were imposed: a short and moderate heatwave at 24 °C for 2 weeks, and a long and intense heatwave with an additional 2 weeks at 26 °C. A positive synergistic effect of heatwaves and nitrate on growth and metabolites was observed, but this also led to increased oxidative stress. In the short and moderate heatwave, oxidative damage was controlled by increased antioxidant levels. The high growth, metabolites, and antioxidants combined with low oxidative stress during the short and moderate heatwaves in moderate nitrate (50 mg L-1) led to a sustainable increased food availability to grazers. On the other hand, long and intense heatwaves in high nitrate conditions caused unsustainable growth due to increased oxidative stress and relatively low antioxidant (proline) levels, increasing the risk for massive algal die-offs.

Synergism Variation between intracellular Glutathione, phycocyanin and SOD in microalgae by carbon quantum dot fluorescence.

Based on the use of CQDs as fluorescent probe and covalent coupling method to detect biological molecules with amino groups, to deeply analysis and detect the metabolism of Microcystis aeruginosa. The metabolic changes of carboxyl biomolecules in Microcystis aeruginosa were analyzed by covalent coupling method, including GSH, phycocyanin and SOD enzyme. The changes of GSH content and its correlation between phycocyanin, SOD were analyzed. The content of phycocyanin and SOD reached the maximum on the 65th day, and GSH was more sensitive to the growth and metabolism of microalgae. GSH plays an important role in reducing the external oxidative damage of microalgae cells. The synthesis of glutathione (GSH), GSH/GSSG mutual transformation, the production of phytochelating peptide (PC), the ASA-GSH cycle, and other physiological processes are interconnected. These interactions are crucial for preserving the antioxidant properties of microalgae and regulating redox-sensitive signal transduction.

Phosphorus limitation combined with aluminum triggers synergistic responses on the freshwater microalgae Raphidocelis subcapitata (Chlorophyceae).

In the environment, algae are exposed to several stressors such as limitation of essential nutrients and excess of toxic substances. It is well known the importance of phosphorus (P) supply for healthy metabolism of algae and impacts at this level can affect the whole aquatic trophic chain. Aluminum (Al) is the most abundant metal on Earth and it is toxic to different trophic levels. Processes related to P and Al assimilation still need to be clarified and little is known about the responses of microalgae exposed to the two stressors simultaneously. We evaluated the effects of environmental concentrations of Al and P limitation, isolated and in combination, on growth, pigment production and photosynthesis of the freshwater microalga Raphidocelis subcapitata. Both stressors affected cell density, chlorophyll a, carotenoids, and maximum quantum yield. Al did not affect any other evaluated parameter, while P limitation affected parameters related to the dissipation of heat by algae and the maximum electron transport rate, decreasing the saturation irradiance. In the combination of both stressors, all parameters evaluated were affected in a synergistic way, i.e., the results were more harmful than expected considering the responses to isolated stressors. Our results indicate that photoprotection mechanisms of algae were efficient in the presence of both stressors, avoiding damages to the photosynthetic apparatus. In addition, our data highlight the higher susceptibility of R. subcapitata to Al in P-limited conditions.

Insight into the roles of hematite iron oxide nanoparticles on microalgae growth, urban wastewater treatment and bioproducts generation: Gompertz simulation, nutrient mass balance and gene expression.

In this study, the effect of α-Fe2O3 nanoparticles spiking in urban wastewater (UWW) on growth rate, wastewater treatment ability and bioproducts generation of C. vulgaris and Spirulina was investigated and compared with pure cultivation system. The biomass concentration of C. vulgaris and Spirulina improved by 20 % and 39 % at 10 and 15 mg/L α-Fe2O3, respectively while the both microalgae growth pattern fitted better with Gompertz simulation after treatment with α-Fe2O3. The nutrients mass balance revealed that 1 g of treated C. vulgaris and Spirulina could uptake more COD, TN and TP in comparison to the untreated cells. The lipid generation increased remarkably (C. vulgaris: 45 % and Spirulina: 72 %) after α-Fe2O3 treatment. While, the addition of α-Fe2O3 showed no significant impact on the protein and carbohydrate productivity. Overall, this study evangelize the role of nanoparticles on promoting microalgae applications as a sustainable approach for UWW treatment and promising feedstock for biofuel production.

Synthetic biology in microalgae towards fucoxanthin production for pharmacy and nutraceuticals.

Synthetic biology has emerged as a powerful tool for engineering biological systems to produce valuable compounds, including pharmaceuticals and nutraceuticals. Microalgae, in particular, offer a promising platform for the production of bioactive compounds due to their high productivity, low land and water requirements, and ability to perform photosynthesis. Fucoxanthin, a carotenoid pigment found predominantly in brown seaweeds and certain microalgae, has gained significant attention in recent years due to its numerous health benefits, such as antioxidation, antitumor effect and precaution osteoporosis. This review provides an overview of the principles and applications of synthetic biology in the microbial engineering of microalgae for enhanced fucoxanthin production. Firstly, the fucoxanthin bioavailability and metabolism in vivo was introduced for the beneficial roles, followed by the biological functions of anti-oxidant activity, anti-inflammatory activity, antiapoptotic role antidiabetic and antilipemic effects. Secondly, the cultivation condition and strategy were summarized for fucoxanthin improvement with low production costs. Thirdly, the genetic engineering of microalgae, including gene overexpression, knockdown and knockout strategies were discussed for further improving the fucoxanthin production. Then, synthetic biology tools of CRISPR-Cas9 genome editing, transcription activator-like effector nucleases as well as modular assembly and chassis engineering were proposed to precise modification of microalgal genomes to improve fucoxanthin production. Finally, challenges and future perspectives were discussed to realize the industrial production and development of functional foods of fucoxanthin from microalgae.

Enzymatic hydrolysis allows an integral valorization of Nannochloropsis oceanica resulting in the production of bioactive peptide extracts and an eicosapentaenoic acid enriched fraction.

Nannochloropsis oceanica is a microalga with relevant protein content, making it a potential source of bioactive peptides. Furthermore, it is also rich in fatty acids, with a special focus on eicosapentaenoic acid (EPA), an omega-3 fatty acid mainly obtained from marine animal sources, with high importance for human health. N. oceanica has a rigid cell wall constraining protein extraction, thus hydrolyzing it may help increase its components' extractability. Therefore, a Box-Behnken experimental design was carried out to optimize the hydrolysis. The hydrolysate A showed 67% ± 0.7% of protein, antioxidant activity of 1166 ± 63.7 µmol TE g-1 of protein and an ACE inhibition with an IC50 of 379 µg protein mL-1 . The hydrolysate B showed 60% ± 1.8% of protein, antioxidant activity of 775 ± 13.0 µmol TE g-1 of protein and an ACE inhibition with an IC50 of 239 µg protein mL-1 . The by-product showed higher yields of total fatty acids when compared to "raw" microalgae, being 5.22% and 1%, respectively. The sustainable developed methodology led to the production of one fraction rich in bioactive peptides and another with interesting EPA content, both with value-added properties with potential to be commercialized as ingredients for different industrial applications, such as functional food, supplements, or cosmetic formulations.

Functional and Extracellular Production and Antitumor Activity of Mouse Alpha Klotho in Model Microalga Chlamydomonas reinhardtii.

Klotho is a human protein with versatile functions associated with longevity and well-being. α-Klotho (α-KL) deficiency in the circulatory system is associated with reduced life expectancy with numerous disorders such as chronic kidney disease, atherosclerosis, infertility, skin atrophy, emphysema, and osteoporosis. The antagonistic effects of Klotho protein against intractable cancers have also been well documented over the past two decades. In addition, recent findings have also illuminated the importance of soluble Klotho during cognitive development, oxidative stress, cellular apoptosis, and neurodegenerative disorders. The low-cost and sustainable production of alpha Klotho protein is extremely important for its widespread use against different diseases. Here, we report heterologous, functional, and extracellular production of mouse α-KL (mα-KL) protein in model microalga Chlamydomonas reinhardtii. The secretion of mα-KL into the extracellular environment facilitated downstream processes and warranted low-cost purification in high-titer. Furthermore, the anticarcinogenic efficiency of recombinant mα-KL was examined and validated on Rattus norvegicus AR42J pancreas tumors. Microalgae-based photosynthetic, low-cost, and scalable production of mα-KL could be used to develop a variety of cosmetics, pharmaceuticals, and wellness products, all aimed at serving health and well-being.

Characterization of chloroplast ribulose-5-phosphate-3-epimerase from the microalga Chlamydomonas reinhardtii.

Carbon fixation relies on Rubisco and 10 additional enzymes in the Calvin-Benson-Bassham cycle. Epimerization of xylulose-5-phosphate (Xu5P) into ribulose-5-phosphate (Ru5P) contributes to the regeneration of ribulose-1,5-bisphosphate, the substrate of Rubisco. Ribulose-5-phosphate-3-epimerase (RPE, EC 5.1.3.1) catalyzes the formation of Ru5P, but it can also operate in the pentose-phosphate pathway by catalyzing the reverse reaction. Here, we describe the structural and biochemical properties of the recombinant RPE isoform 1 from Chlamydomonas (Chlamydomonas reinhardtii) (CrRPE1). The enzyme is a homo-hexamer that contains a zinc ion in the active site and exposes a catalytic pocket on the top of an α8ß8 triose isomerase-type barrel as observed in structurally solved RPE isoforms from both plant and non-plant sources. By optimizing and developing enzyme assays to monitor the reversible epimerization of Ru5P to Xu5P and vice versa, we determined the catalytic parameters that differ from those of other plant paralogs. Despite being identified as a putative target of multiple thiol-based redox modifications, CrRPE1 activity is not affected by both reductive and oxidative treatments, indicating that enzyme catalysis is insensitive to possible redox alterations of cysteine residues. We mapped phosphorylation sites on the crystal structure, and the specific location at the entrance of the catalytic cleft supports a phosphorylation-based regulatory mechanism. This work provides an accurate description of the structural features of CrRPE1 and an in-depth examination of its catalytic and regulatory properties highlighting the physiological relevance of this enzyme in the context of photosynthetic carbon fixation.

Influence of standard culture conditions and effect of oleoresin from the microalga Haematococcus pluvialis on splenic cells from healthy Balb/c mice - a pilot study.

In this work, we used splenocytes from healthy mice to study the effects of the two most commonly used cell culture media (A, B) with different compositions of redox reagents. The incubation of cells for 24 h resulted in a significant decrease in viability and metabolic activity of splenocytes, and the negative effects of incubation in medium B were more pronounced. In standard conditions, oxidative stress in cells was manifested by reduced mitochondrial potential, and this effect correlated with the transition of 58.3% of cells to the early stage of apoptosis under reducing conditions of medium A and up to 66.1% of cells under super-reducing conditions in medium B, suggesting altered cell physiology. High levels of ROS/RNS activated transcription factor Nrf2, superoxide dismutase 1, and catalase. The higher mRNA levels of these genes were under the conditions of medium B, whose super-reducing environment in combination with the environment of conventional incubators proved to be less suitable for the cells compared to medium A. Treatment of the cells with a lower concentration (10 µg/ml) of oleoresin obtained from the microalga H. pluvialis partially eliminated the negative effects of cultivation. Higher concentration of oleoresin (40 µg/ml) was slightly cytotoxic, due to the significant antioxidant effect of astaxanthin, the main bioactive component of the extract, which eliminated most of the ROS/RNS acting as signalling molecules. This study shows that the standard culture conditions do not reflect the physiological in vivo cell conditions; therefore, they are not generally suitable for incubation of all cell types.

Ameliorative effects of Dunaliella salina microalgae on nanoparticle (ZnO NPs)-induced toxicity in fish.

Dunaliella salina (D. salina) is a well-known microalga that contains considerable amounts of nutritious and medicinal bioactive components. This work studied the modulatory role of D. salina against zinc oxide nanoparticle (ZnO NPs)-induced neurotoxic effects in adult zebrafish. Fishes were subjected to 0.69 mg L-1 (1/5th 96-h LC50) for 4 weeks; then, fishes were supplemented with D. salina in the diet for 2 weeks at two levels (15 and 30%). Exposure to ZnO NPs induced a significant increase in the levels of reactive oxygen species (ROS), hydrogen peroxide (H2O2), malondialdehyde (MDA), and 8-hydroxy-2-deoxyguanosine (8-OH-dG) while accompanied with downregulation of antioxidant genes in the brain of exposed fishes. Brain neurochemistry and enzyme activities were also altered following ZnO NP exposure. ZnO NPs significantly reduced the neurotransmitters and acetylcholinesterase (AchE) activity while increasing Alzheimer's disease-related proteins and inflammatory response via upregulation of tumor necrosis factor (TNF-α). Additionally, ZnO NPs increased the indices of brain's DNA oxidative damage, increasing brain tissue's metallothionein (MT) and zinc residues. ZnO NPs upregulated the transcription patterns of apoptosis-related genes (casp3 and p53). D. salina dietary co-supplementation with ZnO NPs alleviated the ZnO NPsZnO NP-induced neuro-oxidative damages by lowering the lipid, DNA damage, and inflammatory biomarkers. Besides, D. salina alleviating responses were linked with increasing the levels of the assessed antioxidants. Conclusively, D. salina dietary supplementation induced potential alleviating effects of the ZnO NP-induced neurotoxicity in adult zebrafish.

Effects of phytohormone on Chlorella vulgaris grown in wastewater-flue gas: C/N/S fixation, wastewater treatment and metabolome analysis.

Chlorella vulgaris (C. vulgaris) can provide the means to fix CO2 from complicated flue gas, treat wastewater and reach a sustainable production of petrochemical substitutes simultaneously. However, a prerequisite to achieving this goal is to promote C. vulgaris growth and improve the CO2-to-fatty acids conversion efficiency under different conditions of flue gas and wastewater. Thus, the addition of indole-3-acetic acid (IAA) in C. vulgaris cultivation was proposed. Results showed that C. vulgaris were more easily inhibited by 100 ppm NO and 200 ppm SO2 under low nitrogen (N) condition. NO and SO2 decreased the carbon (C) fixation; but increased N and sulfur (S) fixation. IAA adjusted the content of superoxide dismutase (SOD) and malondialdehyde (MDA), improved the expression of psbA, rbcL, and accD, attenuated the toxicity of NO and SO2 on C. vulgaris, and ultimately improved cell growth (2014.64-2458.16 mgdw·L-1) and restored CO2 fixation rate (170.98-220.92 mg CO2·L-1·d-1). Moreover, wastewater was found to have a high treatment efficiency because C. vulgaris grew well in all treatments, and the maximal removal rates of both N and phosphorus (P) reached 100%. Metabonomic analysis showed that IAA, "NO and SO2" were involved in the down-regulated and up-regulated expression of multiple metabolites, such as fatty acids, amino acids, and carbohydrates. IAA was beneficial for improving lipid accumulation with 24584.21-27634.23 µg g-1, especially monounsaturated fatty acids (MUFAs) dominated by 16-18 C fatty acids, in C. vulgaris cells. It was concluded that IAA enhanced the CO2 fixation, fatty acids production of C. vulgaris and its nutrients removal rate.

The Photoprotective Protein PsbS from Green Microalga Lobosphaera incisa: The Amino Acid Sequence, 3D Structure and Probable pH-Sensitive Residues.

PsbS is one of the key photoprotective proteins, ensuring the tolerance of the photosynthetic apparatus (PSA) of a plant to abrupt changes in irradiance. Being a component of photosystem II, it provides the formation of quenching centers for excited states of chlorophyll in the photosynthetic antenna with an excess of light energy. The signal for "turning on" the photoprotective function of the protein is an excessive decrease in pH in the thylakoid lumen occurring when all the absorbed light energy (stored in the form of transmembrane proton potential) cannot be used for carbon assimilation. Hence, lumen-exposed protonatable amino acid residues that could serve as pH sensors are the essential components of PsbS-dependent photoprotection, and their pKa values are necessary to describe it. Previously, calculations of the lumen-exposed protonatable residue pKa values in PsbS from spinach were described in the literature. However, it has recently become clear that PsbS, although typical of higher plants and charophytes, can also provide photoprotection in green algae. Namely, the stress-induced expression of PsbS was recently shown for two green microalgae species: Chlamydomonas reinhardtii and Lobosphaera incisa. Therefore, we determined the amino acid sequence and modeled the three-dimensional structure of the PsbS from L. incisa, as well as calculated the pKa values of its lumen-exposed protonatable residues. Despite significant differences in amino acid sequence, proteins from L. incisa and Spinacia oleracea have similar three-dimensional structures. Along with the other differences, one of the two pH-sensing glutamates in PsbS from S. oleracea (namely, Glu-173) has no analogue in L. incisa protein. Moreover, there are only four glutamate residues in the lumenal region of the L. incisa protein, while there are eight glutamates in S. oleracea. However, our calculations show that, despite the relative deficiency in protonatable residues, at least two residues of L. incisa PsbS can be considered probable pH sensors: Glu-87 and Lys-196.