Eco-friendly cultivation of microalgae using a horizontal twin layer system for treatment of real solid waste leachate.

Solid waste leachate (SWL) requires dilution with water to offset the negative effects of high nutrient concentration and organic compounds for its microalgae-based treatment. Among attached cultivation systems, twin layer is a technology in which limited information is available on treatment of high strength wastewater using microalgae. Moreover, widespread application of twin layer technology is limited due to cost of substrate and source layer used. In the present study, potential of Scenedesmus sp. for the treatment of SWL was assessed on horizontal twin layer system (HTLS). Novel and cost-effective substrate layers were tested as attachment material. Wetland treated municipal wastewater (WMW) was used to prepare SWL dilutions viz, 5%, 10%, 15%, 20% and 25% SWL. Recycled printing paper showed maximum biomass productivity of 5.19 g m-2 d-1. Among all the SWL dilutions, Scenedesmus sp. achieved maximum growth of 103.05 g m-2 in 5% SWL which was 16% higher than WMW alone. The maximum removal rate of NH4+ -N, TKN, and PO43- P was obtained in 20% SWL which was 1371, 1588 and 153 mg m-2 d-1 respectively. Varying concentrations of nutrients in different SWL dilutions significantly affected lipid biosynthesis, with enhanced productivity of 2.28 g m-2 d-1 achieved in 5% SWL compared to 0.97 g m-2 d-1 in 20% SWL. Hence, it can be concluded that 5% SWL dilution was good for biomass and lipid production, while the highest nutrient removal rates were obtained at 20% SWL mainly attributed to biotic and abiotic processes. Based on these results HTLS can be a promising technology for pilot scale to explore industrialized application of wastewater treatment and algal production.

Efficient removal of thiamethoxam by freshwater microalgae Scenedesmus sp. TXH: Removal mechanism, metabolic degradation and application.

Neonicotinoids, as the most widely used pesticides in the world, help improve the production of crops. Meanwhile, it also brings potential threats to surrounding environments and other organisms because of its wide use and even abuse. In this study, Scenedesmus sp. TXH isolated from a wastewater treatment plant was used to remove the neonicotinoid pesticide thiamethoxam (THIA). The removal efficiency, degradation pathway, metabolite fate of THIA and physicochemical effects on microalgae cells were studied. Meanwhile, the feasibility of using microalgal technology to remove THIA from municipal wastewater was also explored. The results showed that 5-40 mg/L of THIA slightly promoted the growth of microalgae, while 60 mg/L THIA severely inhibited microalgal growth. It was observed that malondialdehyde content and superoxide dismutase activity in 60 mg/L THIA group increased significantly (p < 0.05) in the early stage of the experiment, indicating that THIA caused oxidative damage to microalgae. Scenedesmus sp. TXH showed high-efficient degradation ability and high resistance to THIA, with 100% removal of THIA at 5, 20 and 40 mg/L groups and 97.5% removal of THIA at 60 mg/L group on day 12. THIA was mainly removed by biodegradation, accounting for 78.18%, 93.50%, 96.81% and 91.35% under 5, 20, 40 and 60 mg/L on day 12, respectively. Six degradation products were identified, and four potential degradation pathways were proposed. In practical wastewater, the removal efficiency of total dissolved nitrogen, total dissolved phosphorus, ammonia nitrogen and THIA reached 85.68%, 90.00%, 98.43% and 100%, respectively, indicating that Scenedesmus sp. TXH was well adapted to the wastewater and effectively removed THIA and conventional pollutants.

Distribution of the Water-Soluble Astaxanthin Binding Carotenoprotein (AstaP) in Scenedesmaceae.

Photooxidative stress-inducible water-soluble astaxanthin-binding proteins, designated as AstaP, were identified in two Scenedesmaceae strains, Coelastrella astaxanthina Ki-4 and Scenedesmus obtusus Oki-4N; both strains were isolated under high light conditions. These AstaPs are classified as a novel family of carotenoprotein and are useful for providing valuable astaxanthin in water-soluble form; however, the distribution of AstaP orthologs in other microalgae remains unknown. Here, we examined the distribution of AstaP orthologs in the family Scenedesmaceae with two model microalgae, Chlamydomonas reinhardtii and Chlorella variabilis. The expression of AstaP orthologs under photooxidative stress conditions was detected in cell extracts of Scenedesmaceae strains, but not in model algal strains. Aqueous orange proteins produced by Scenedesmaceae strains were shown to bind astaxanthin. The protein from Scenedesmus costatus SAG 46.88 was purified. It was named ScosAstaP and found to bind astaxanthin. The deduced amino acid sequence from a gene encoding ScosAstaP showed 62% identity to Ki-4 AstaP. The expression of the genes encoding AstaP orthologs was shown to be inducible under photooxidative stress conditions; however, the production amounts of AstaP orthologs were estimated to be approximately 5 to 10 times lower than that of Ki-4 and Oki-4N.

Effect of olive mill wastewaters on Scenedesmus sp. growth, metabolism and polyphenols removal.

BACKGROUND: The three-phase extraction process of olive oil produces highly contaminated wastewater (OMW). The elimination of this toxic by-product is an important environmental issue that requires the development of an appropriate management solution. The cultivation of microalgae using OMW as growth medium was therefore studied using single (the culture medium was formed by 0% to 80% ultrafiltered olive mill wastewater (OMUF) or OMW added to BG11) and two-stage strategies (microalgae were firstly cultivated in the BG11 medium. In the second stage, 40% and 80% of OMUF and OMW were added to the culture). In this work, biodegradation of OMW and subsequent extraction of lipid and antioxidant molecules was investigated as an ecofriendly method for the bioremediation and valorization of OMW. RESULTS: For two-stage cultivation, OMUF and OMW stress enhanced the intracellular amount of polyphenol accumulated in Scenedesmus sp. and exhibited the highest 2, 2-diphenyl-1- picrylhydrazyl radical (DPPH) and 2,2'-azino-bis (3-ethylbenzoline-6-sulfonate) radical (ABTS) scavenging ability compared with single-stage cultivation. Moreover, the lipid profile is dominated by polyunsaturated acids. In the single-stage cultivation, the Ch a, Ch b, carotenoid, carbohydrate and lipid content of 2.57, 7.4, 1.69, 368, and 644 g kg-1 were observed in 40% OMUF added culture, respectively, along with high biomass productivity and 58% of polyphenol removal. Moreover, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that the biomass of Scenedesmus sp. cultured on 40% OMUF did not show any toxic effect, making it an efficient strategy. CONCLUSION: The results indicate that Scenedesmus sp. is a promising microalga for the biotreatment of OMW and the extraction of bioactive metabolites. © 2021 Society of Chemical Industry.

Structural elucidation of selenocysteine insertion machinery of microalgal selenoprotein T and its transcriptional analysis.

Essential trace element selenium in association with selenoproteins, which is found in almost all organisms except higher plants and fungi, is involved in various biological functions. Advancement in the field of whole genome sequencing and data analyzing bioinformatic tools led to the accumulation of genome information of organisms. However, selenoproteins are unique and it needs specialized genomics tool for its identification as well as characterization. In this study, the presence of selenoprotein T (SelT) from Scenedesmus quadricauda was shown for the first time with experimental evidence and compared with SelT of marine microalgae Nannochloropsis oceanica. Along with SelT, all the associated machineries required to synthesize the selenoproteins were also identified. Also, the present study tried to explicate the evolutionary relatedness of SelT of these two organisms with other known bacteria and eukaryotes. Transcript level analysis in S. quadricauda under endoplasmic reticulum stress showed a 1.2 ± 0.28-fold increase in SelT expression. Thus, it provided the first experimental evidence on SelT expression from microalgae.

Microbial lipid production by oleaginous yeasts grown on Scenedesmus obtusiusculus microalgae biomass hydrolysate.

Due to increasing oil prices and climate change concerns, biofuels have become increasingly important as potential alternative energy sources. However, the use of arable lands and valuable resources for the production of biofuel feedstock compromises food security and negatively affect the environment. Single cell oils (SCOs), accumulated by oleaginous yeasts, show great promise for efficient production of biofuels. However, the high production costs attributed to feedstocks or raw materials present a major limiting factor. The fermentative conversion of abundant, low-value biomass into microbial oil would alleviate this limitation. Here, we explore the feasibility of utilizing microalgae-based cell residues as feedstock for yeast oil production. We developed an efficient, single-step enzymatic hydrolysis to generate Scenedesmus obtusiusculus hydrolysate (SH) without thermo-chemical pretreatment. With this eco-friendly process, glucose conversion efficiencies reached 90-100%. Cutaneotrichosporon oleaginosus, Cryptococcus curvatus and Rhodosporidium toruloides were cultivated on SH as sole nutrients source. Only C. oleaginosus was able to accumulate intracellular lipids, with a 35% (g lipid/g DCW) content and a yield of 3.6 g/L. Our results demonstrate the potential valorization of algal biomass into desired end-products such as biofuels.

Effect of cell disruption methods on the extraction of bioactive metabolites from microalgal biomass.

Microalgae synthesize a variety of potentially high-value compounds. Due to their robust cell wall, cell disruption is necessary to improve extraction of these compounds. While cell disruption methods have been optimized for lipid and protein extraction, there are limited studies for other bioactive compounds. The present study investigated the effect of freeze-drying combined with sonication or ball-milling on the extraction of antioxidant and plant biostimulating compounds from Chlorella sp., Chlorella vulgaris and Scenedesmus acutus. Both cell disruption methods resulted in higher extract yields from the biomass compared to freeze-dried biomass using 50% methanol as a solvent. Antioxidant activity of Chlorella extracts was generally higher than freeze-dried extracts based on the diphenylpicrylhydrazyl (DPPH) and ß-carotene linoleic acid assays. However, the effectiveness of each treatment varied between microalgae strains. Sonication resulted in the highest antioxidant activity in Chlorella sp. extracts. Ball-milling gave the best results for C. vulgaris extracts in the DPPH assay. Both cell disruption methods decreased antioxidant activity in S. acutus extracts. Plant biostimulating activity was tested using the mung bean rooting assay. Damaging the membrane by freeze-drying was sufficient to release the active compounds using water extracts. In contrast, both cell disruption methods negatively affected the biological activity of the extracts. These results indicate that bioactive compounds in microalgae are sensitive to post-harvest processes and their biological activity can be negatively affected by cell disruption methods. Care must be taken to not only optimize yield but to also preserve the biological activity of the target compounds.

Dietary Scenedesmus ovalternus improves disease resistance of overwintering gibel carp (Carassius gibelio) by alleviating toll-like receptor signaling activation.

This study was conducted to investigate the effect of dietary Scenedesmus ovalternus on the growth and disease resistance of gibel carp (Carassius gibelio) during overwintering. Gibel carp (initial body weight: 90.39 ± 0.33 g) were fed with diets containing 0% or 4% Scenedesmus ovalternus (DS0 and DS4) for 4 weeks during the early overwintering period, and then all fish were left unfed during the late overwintering period. A bacterial challenge test using Aeromonas hydrophila was subsequently conducted. The 4% Scenedesmus ovalternus diet had no effect on the growth of gibel carp (P > 0.05), but did improve the survival rate after the challenge (P ≤ 0.05). In the DS0 group, the bacterial challenge decreased the contents of complement 3 (C3), immunoglobulin M (IgM), interleukin 2 (IL2) and tumor necrosis factor α (TNFα) in fish (P < 0.05); in the DS4 group, the challenge increased total antioxidant capacity (T-AOC) and myeloperoxidase (MPO) activity but decreased IL2 and TNFα contents (P < 0.05). The activities of MPO and contents of C3, IgM and TNFα were higher in the DS4 group than that fed the DS0 diet after bacterial challenge (P < 0.05). Compared to pre challenge, the expression levels of toll like receptor 2 (TLR2), toll like receptor 3 (TLR3), toll like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain-containing adaptor protein (TIRAP), TIR-domain-containing adapter-inducing interferon ß (TRIF), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor α (IκBα), transforming growth factor ß (TGFß), interleukin 1ß (IL1ß), tumor necrosis factor α1 (TNFα1) and interleukin 10 (IL10) in the head kidney of gibel carp were induced after challenge (P < 0.05). Gibel carp fed the DS4 diet showed lower expression of TGFß in head kidney before the challenge and lower expression of TLR2, TLR3, TLR4, TIRAP, TRIF, IκBα, TNFα1, IL10 and TGFß after the challenge than that fed the DS0 diet (P < 0.05). Overall, Scenedesmus ovalternus supplement enhanced the resistances of gibel carp against A. hydrophila after overwintering via the TLR signaling pathway.

Effect of nitrogen deficiency on the physiology and biochemical composition of microalga Scenedesmus rotundus-MG910488.

The present investigation ascertains the impact of gradient concentrations of sodium nitrate on the physiology and biochemical composition of isolated microalga Scenedesmus rotundus-MG910488. The concentrations of nitrate were selected as 0, 3.5, 7.0, 10.5, 14.0, and 17.6 mM/L in BG11 medium. The lower concentrations of nitrogen were found to be significantly decreasing the cell count and photosynthetic activity in the microalga as well as changing cell morphology. The amount of biomass, its productivity and lipid yield were significantly affected. The highest biomass of 689.15 ± 14.27 mg/L was achieved in the concentration of 17.6 mM/L with the biomass productivity of 38.28 ± 0.78 mg/L. The highest lipid accumulation of 41.46 ± 1.94% dry-cell weight was obtained at a concentration of 3.5 mM/L, whereas the lowest lipid accumulation of 29.22 ± 1.65% at the concentration of 17.6 mM/L sodium nitrate. The fatty acid composition determines the quality of the fuel, so the characterization of fatty acid methyl esters (FAMEs) was performed by GC, and the assessment of methyl esters of fatty acid confirmed the existence of palmitic acid, oleic acid, and linoleic acid, which are essential components suitable for biodiesel production. FTIR confirms the presence of FAME components by estimating the bending and stretching of functional groups.

Novel protocol optimized for microalgae lutein used as food additives.

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.

Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA).

The analysis of protein in algal biomass is one of the most critical areas of commercial development of algae characterization for nutritional or other high value applications. A new rapid and accurate method is required that can be widely implemented and that is free from interferences from the complex algal biomass matrix. We developed a simple spectrophotometric method for primary amino acid quantification bulk measurement in an acid hydrolyzed algal biomass preparation, as an alternative to the more labor-intensive amino HPLC acid analysis or less specific nitrogen-to-protein conversion. We have validated an O-phthalaldehyde (OPA)-based derivatization method, showing accurate and linear quantification for standard reference amino acids as well as mixtures, mimicking the amino acid complexity found in algal biomass. The presence of interferences that may be derived from the complex biomass biochemical composition was tested during the method validation phase. We document the application of a novel method of OPA derivatization with 3-mercaptopropionic acid (3MPA) to determine the total amino acid content of harvested algal biomass collected from different, controlled cultivation conditions and demonstrated a within 10% accuracy against a reference measurement of amino acid content in at least 4 species and 10 algal biomass samples, across early, mid, and late-stages of cultivation.

Supercritical Fluid Extraction of Lutein from Scenedesmus almeriensis.

Lutein has several benefits for human health, playing an important role in the prevention of age-related macular degeneration (AMD), cataracts, amelioration of the first stages of atherosclerosis, and some types of cancer. In this work, the Scenedesmus almeriensis microalga was used as a natural source for the supercritical fluid (SF) extraction of lutein. For this purpose, the optimization of the main parameters affecting the extraction, such as biomass pre-treatment, temperature, pressure, and carbon dioxide (CO2) flow rate, was performed. In the first stage, the effect of mechanical pre-treatment (diatomaceous earth (DE) and biomass mixing in the range 0.25-1 DE/biomass; grinding speed varying between 0 and 600 rpm, and pre-treatment time changing from 2.5 to 10 min), was evaluated on lutein extraction efficiency. In the second stage, the influence of SF-CO2 extraction parameters such as pressure (25-55 MPa), temperature (50 and 65 °C), and CO2 flow rate (7.24 and 14.48 g/min) on lutein recovery and purity was investigated. The results demonstrated that by increasing temperature, pressure, and CO2 flow rate lutein recovery and purity were improved. The maximum lutein recovery (~98%) with purity of ~34% was achieved operating at 65 °C and 55 MPa with a CO2 flow rate of 14.48 g/min. Therefore, optimum conditions could be useful in food industries for lutein supplementation in food products.

Comparative biodegradation of all chlorinated phenols by the microalga Scenedesmus obliquus - The biodegradation strategy of microalgae.

This work presents the comparative biodegradation of all chlorinated phenolic compounds by the green alga Scenedesmus obliquus and determines the microalgal bioenergetic strategy. The microalga manages its energy reserves rationally by investing them, either on growth or on the biodegradation of the toxic compound. The microalga seems to follow two distinct detoxification strategies. In the first one, when microalgae are surrounded by relatively low toxic phenolic compounds (phenol, monochlorophenols, 2,4-dichlorophenol and 2,6-dichlorophenol), they use all, or at least more of their energy reserves to increase the biomass production and not the biodegradation. In the second one, when surrounded by higher toxic chlorophenols (meta-substituted dichlorophenols, trichlorophenols, tetrachlorophenols and pentachlorophenol) the microalgae invest more, or all of their energy reserves directly in the biodegradation of the toxic compounds, while less or no energy is invested in biomass increase. The microalga biodegraded in five days approximately 9% of the lower toxic phenol and 90% of the higher toxic pentachlorophenol. Considering our ability to interfere with microalgae energy reserves, which define their stress tolerance in the toxic environment, and knowing the microalgal bioenergetic strategy, we could easily use microalgae to biodegrade toxic wastes in the frame of a rational biotechnological approach in the near future.

Influences of carbon and nitrogen sources and metal ions on the heterotrophic culture of Scenedesmus sp. LX1.

In this study, the influences of organic carbon sources (OCS, including xylose, glucose, maltose, sucrose, and starch) and inorganic and organic nitrogen sources (INS, including ammonia chloride and sodium nitrate; ONS, including arginine, alanine, proline, and valine) and metal ions (including Na+, K+, Mn2+, Zn2+ and Cu2+) on the growth, lipid accumulation, and nitrogen and phosphorus (N&P) removal capabilities of oleaginous Scenedesmus sp. LX1 under heterotrophic conditions were investigated. The results showed that glucose was the only OCS for Scenedesmus sp. LX1 to grow well with specific growth rate of 0.935 days-1, maximum biomass of 1.72 g L-1, and largest removal rates of N&P and organic carbon reaching 72.228%, 93.034%, and 19.208%, respectively. After 11 days of cultivation, the maximal biomass reached in the group with starch or glucose while maximal lipid and triacylglycerol (TAG) yields reached in the groups with maltose and sucrose, respectively. Sodium nitrate was best nitrogen source as the largest algal density, maximal yields of lipids and TAGs, and highest N&P removal rates reached up to 1.105 × 107 cells·mL-1, 196.70 mg L-1, 5.19 mg L-1, 89.61% and 100%, respectively. Scenedesmus sp. LX1 was found to have great tolerance to Na+, K+, Mn2+, and Zn2+ while 0.5 mg L-1 Cu2+ had a strong inhibition on growth and N&P removal rate of Scenedesmus sp. LX1. Concentration increasing of five metal ions all caused the yield increases of microalgal lipid and TAGs. Graphical abstract.

Carbohydrates and polyphenolics of extracts from genetically altered plant acts as catalysts for in vitro synthesis of silver nanoparticle.

Eco-friendly biosynthetic approach for silver nanoparticles production using plant extracts is an exciting advancement in bio-nanotechnology and has been successfully attempted in nearly 41 plant species. However, an established model plant system for systematically unraveling the biochemical components required for silver nanoparticles production is lacking. Here we used Arabidopsis thaliana as the model plant for silver nanoparticles biosynthesis in vitro. Employing biochemical, spectroscopic methods, selected mutants and over-expressor plants of Arabidopsis involved in pleotropic functions and sugar homeostasis, we show that carbohydrates, polyphenolics and glyco-proteins are essential components which stimulated silver nanoparticles synthesis. Using molecular genetics as a tool, our data enforces the requirement of sugar conjugated proteins as essentials for AgNPs synthesis over protein alone. Additionally, a comparative analysis of AgNPs synthesis using the aqueous extracts of some of the plant species found in a brackish water ecosystem (Gracilaria, Potamogeton, Enteromorpha and Scendesmus) were explored. Plant extract of Potamogeton showed the highest potential of nanoparticles production comparable to that of Arabidopsis among the species tested. Silver nanoparticles production in the model plant Arabidopsis not only opens up a possibility of using molecular genetics tool to understand the biochemical pathways and components in detail for its synthesis.

Microalga Scenedesmus bajacalifornicus BBKLP-07, a new source of bioactive compounds with in vitro pharmacological applications.

Microalgae are photosynthetic eukaryotes which are primary producers in the food chain and also excellent sources for bioactive compounds such as alkaloids, flavonoids, phenols, saponins and other fine chemicals. In the present study, the microalga Scenedesmus bajacalifornicus BBKLP-07 was subjected to soxhlet extraction using solvents like chloroform, acetone, ethanol, methanol and aqueous solvents. All the solvents were tested for the presence of phytochemical constituents such as alkaloids, flavonoids, glycosides, phenols, lignin's, saponins, sterols, tannins, anthraquinone and reducing sugar using the standard procedures. Furthermore, all the crude extracts were subjected to antidiabetic, antioxidant, anti-inflammatory and antimicrobial activities. Antidiabetic activity of the microalgal extracts was observed maximum in Aqueous extract. Methanolic extracts have shown maximum antioxidant activity and chloroform extracts have exhibited highest anti-inflammatory effects. Antimicrobial activities were tested against E.coli, S, typhi, C.perfringens and B.subtilis bacteria and fungi A.niger, and C. albicans. Therefore, the green microalga Scenedesmus bajacalifornicus BBKLP-07 is a rich source of biological active compounds and nutraceuticals and can be exploited for commercial applications.

Evaluation of antioxidant and antibacterial capacity of green microalgae Scenedesmus subspicatus.

Microalgae are considered one of the most promising raw materials for the development of high value products for pharmaceuticals, nutraceuticals, and cosmetic industries, as well as being potential sources of protein, vitamins, and minerals for human consumption. Hence, the present research focuses extraction of antioxidant and antimicrobial compounds from Scenedesmus subspicatus using solvents of different polarities. Different solvents such as ethanol, methanol, butanol, acetone, dimethyl sulfoxide, and water were used to extract compounds from the green microalgae S. subspicatus and then they were examined for phytochemical screening, antioxidant activity, and antimicrobial properties. In vitro free radical quenching and total antioxidant activity of extracts were investigated with 1,1-diphenyl-2-picryl hydrazyl and compared with catequin and gallic acid as positive controls. The antimicrobial activity was evaluated in gram-negative and gram-positive bacteria. Aqueous extracts and dimethyl sulfoxide presented better performance in phytochemical analysis. This result showed consistency in the sequential tests. The antioxidant activity was also better using the two solvents cited above. The extracts acetone, water, and dimethyl sulfoxide showed ability to inhibit the growth of Bacillus subtilis. However, only dimethyl sulfoxide inhibited the growth of Klebsiella pneumoniae and Escherichia coli. Use of the aqueous extract, proven its effectiveness, is an economic protocol and avoids the use of toxic substances.

Photosynthetic physiological performance and proteomic profiling of the oleaginous algae Scenedesmus acuminatus reveal the mechanism of lipid accumulation under low and high nitrogen supplies.

In this study, we presented cellular morphological changes, time-resolved biochemical composition, photosynthetic performance and proteomic profiling to capture the photosynthetic physiological response of Scenedesmus acuminatus under low nitrogen (3.6 mM NaNO3, N-) and high nitrogen supplies (18.0 mM NaNO3, N+). S. acuminatus cells showed extensive lipid accumulation (53.7% of dry weight) and were enriched in long-chain fatty acids (C16 & C18) under low nitrogen supply. The activity of PSII and photosynthetic rate decreases, whereas non-photochemical quenching and dark respiration rates were increased in the N- group. In addition, the results indicated a redistribution of light excitation energy between PSII and PSI in S. acuminatus exists before lipid accumulation. The iTRAQ results showed that, under high nitrogen supply, protein abundance of the chlorophyll biosynthesis, the Calvin cycle and ribosomal proteins decreased in S. acuminatus. In contrast, proteins associated with the photosynthetic machinery, except for F-type ATPase, were increased in the N+ group (N+, 3 vs. 9 days and 3 days, N+ vs. N-). Under low nitrogen supply, proteins involved in central carbon metabolism, fatty acid synthesis and branched-chain amino acid metabolism were increased, whereas the abundance of proteins of the photosynthetic machinery had decreased, with exception of PSI (N-, 3 vs. 9 days and 9 days, N+ vs. N-). Collectively, the current study has provided a basis for the metabolic engineering of S. acuminatus for biofuel production.

Characteristics of intracellular algogenic organic matter and its reactivity with hydroxyl radicals.

The aim of this study was to investigate the reactivity of intracellular algogenic organic matter (IOM) with hydroxyl radicals (·OH), a key reaction species in advanced oxidation processes. IOM was extracted from two green algae, Chlamydomonas reinhardtii and Scenedesmus sp., and two blue-green algae, Anabaena sp. and Microcystis aeruginosa using a freeze-thaw method. The second-order rate constants of the extracted IOM with ·OH were determined as 7.95 × 108 MC-1 s-1 (Chlamydomonas reinhardtii), 6.71 × 108 MC-1 s-1 (Scenedesmus sp.), 4.02 × 108 MC-1 s-1 (Anabaena sp.), and 4.45 × 108 MC-1 s-1 (Microcystis aeruginosa). These rate constants were significantly higher than values reported for dissolved organic matter in various water sources. This implies that IOM formation during algal bloom season could change the ·OH water matrix demand and adversely affect the performance of advanced oxidation processes. To investigate the physical and chemical composition characteristics of IOM and their relationship to the rate constants determined for the reaction between IOM and ·OH, liquid chromatography-organic carbon detection (LC-OCD) and fluorescence excitation-emission matrix & parallel factor analysis (FEEM-PARAFAC) were used. The IOM mainly consisted of low molecular weight (LMW) matter and protein-related compounds, as evidenced LMW neutrals (38-65%), biopolymers (7-19%), and tryptophan-like compounds (74-94%). Based on the composition characteristics of IOM, it was concluded that the molecular weight and the presence of nitrogen-containing compounds are influential parameters for determining the reactivity of IOM with ·OH.

Bilirubin detoxification using different phytomaterials: characterization and in vitro studies.

BACKGROUND: Activated carbon (AC) is a common adsorbent that is used in both artificial and bioartificial liver devices. METHODS: Three natural materials - date pits of Phoenix dactylifera (fruit), Simmondsia chinensis (jojoba) seeds, and Scenedesmus spp. (microalgae) - were used in the present investigation as precursors for the synthesis of AC using physical activation. The chemical structures and morphology of AC were analyzed. Then, AC's bilirubin adsorption capacity and its cytotoxicity on normal liver (THLE2) and liver cancer (HepG2) cells were characterized. RESULTS: Compared with the other raw materials examined, date-pit AC was highly selective and showed the most effective capacity of bilirubin adsorption, as judged by isotherm-modeling analysis. MTT in vitro analysis indicated that date-pit AC had the least effect on the viability of both THLE2 and HepG2 cells compared to jojoba seeds and microalgae. All three biomaterials under investigation were used, along with collagen and Matrigel, to grow cells in 3D culture. Fluorescent microscopy confirmed date-pit AC as the best to preserve liver cell integrity. CONCLUSION: The findings of this study introduce date-pit-based AC as a novel alternative biomaterial for the removal of protein-bound toxins in bioartificial liver devices.