Microalgae: A Promising Source of Bioactive Phycobiliproteins.

Phycobiliproteins are photosynthetic light-harvesting pigments isolated from microalgae with fluorescent, colorimetric and biological properties, making them a potential commodity in the pharmaceutical, cosmetic and food industries. Hence, improving their metabolic yield is of great interest. In this regard, the present review aimed, first, to provide a detailed and thorough overview of the optimization of culture media elements, as well as various physical parameters, to improve the large-scale manufacturing of such bioactive molecules. The second section of the review offers systematic, deep and detailed data about the current main features of phycobiliproteins. In the ultimate section, the health and nutritional claims related to these bioactive pigments, explaining their noticeable potential for biotechnological uses in various fields, are examined.

Structural Characterization and Rheological and Antioxidant Properties of Novel Polysaccharide from Calcareous Red Seaweed.

A novel sulfated xylogalactan (JASX) was extracted and purified from the rhodophyceae Jania adhaerens. JASX was characterized by chromatography (GC/MS-EI and SEC/MALLS) and spectroscopy (ATR-FTIR and 1H/13C NMR) techniques. Results showed that JASX was constituted by repeating units of (→3)-ß-d-Galp-(1,4)-3,6-α-l-AnGalp-(1→)n and (→3)-ß-d-Galp-(1,4)-α-l-Galp-(1→)n substituted on O-2 and O-3 of the α-(1,4)-l-Galp units by methoxy and/or sulfate groups but also on O-6 of the ß-(1,3)-d-Galp mainly by ß-xylosyl side chains and less by methoxy and/or sulfate groups. The Mw, Mn, D, [η] and C* of JASX were respectively 600 and 160 kDa, 3.7, 102 mL.g-1 and 7.0 g.L-1. JASX exhibited pseudoplastic behavior influenced by temperature and monovalent salts and highly correlated to the power-law model and the Arrhenius relationship. JASX presented thixotropic characteristics, a gel-like viscoelastic behavior and a great viscoelasticity character. JASX showed important antioxidant activities, outlining its potential as a natural additive to produce functional foods.

Molecular and Structural Characterizations of Lipases from Chlorella by Functional Genomics.

Microalgae have been poorly investigated for new-lipolytic enzymes of biotechnological interest. In silico study combining analysis of sequences homologies and bioinformatic tools allowed the identification and preliminary characterization of 14 putative lipases expressed by Chlorella vulagaris. These proteins have different molecular weights, subcellular localizations, low instability index range and at least 40% of sequence identity with other microalgal lipases. Sequence comparison indicated that the catalytic triad corresponded to residues Ser, Asp and His, with the nucleophilic residue Ser positioned within the consensus GXSXG pentapeptide. 3D models were generated using different approaches and templates and demonstrated that these putative enzymes share a similar core with common α/ß hydrolases fold belonging to family 3 lipases and class GX. Six lipases were predicted to have a transmembrane domain and a lysosomal acid lipase was identified. A similar mammalian enzyme plays an important role in breaking down cholesteryl esters and triglycerides and its deficiency causes serious digestive problems in human. More structural insight would provide important information on the enzyme characteristics.

Insight into Arthrospira platensis Δ9 desaturase: a key enzyme in poly-unsaturated fatty acid synthesis.

Membrane-bound Δ9 desaturase perform oxygenated desaturation reactions to insert the first double bonds within fatty acyl chains between C9 and C10 positions of most saturated substrates. Arthrospira platensis, a blue green microalga, is an important source of polyunsaturated fatty acids (PUFA) such as oleic, linoleic and linolenic acids lending benefits and functions in dietetics and therapeutic uses. In this paper, we report homology modeling and docking studies of a Δ9 desaturase from Arthrospira platensis strain. The protein model showed high topology resemblance compared to membrane-bound desaturases with a cytoplasmic core displaying the catalytic site and a transmembrane domain created by four α-helices. The cytoplasmic cap contained the three conserved-histidine boxes typical for all membrane bound desaturases. The protein model was used to perform protein-protein docking and the dimer structure was generated. The two monomers are tightly related with hydrophobic interactions between the transmembrane domain helices. The study highlighted also the potent role of a particular 53 residues sequence located at the N terminal end of the enzyme.

First study of correlation between oleic acid content and SAD gene polymorphism in olive oil samples through statistical and bayesian modeling analyses.

BACKGROUND: Virgin olive oil is appreciated for its particular aroma and taste and is recognized worldwide for its nutritional value and health benefits. The olive oil contains a vast range of healthy compounds such as monounsaturated free fatty acids, especially, oleic acid. The SAD.1 polymorphism localized in the Stearoyl-acyl carrier protein desaturase gene (SAD) was genotyped and showed that it is associated with the oleic acid composition of olive oil samples. However, the effect of polymorphisms in fatty acid-related genes on olive oil monounsaturated and saturated fatty acids distribution in the Tunisian olive oil varieties is not understood. METHODS: Seventeen Tunisian olive-tree varieties were selected for fatty acid content analysis by gas chromatography. The association of SAD.1 genotypes with the fatty acids composition was studied by statistical and Bayesian modeling analyses. RESULTS: Fatty acid content analysis showed interestingly that some Tunisian virgin olive oil varieties could be classified as a functional food and nutraceuticals due to their particular richness in oleic acid. In fact, the TT-SAD.1 genotype was found to be associated with a higher proportion of mono-unsaturated fatty acids (MUFA), mainly oleic acid (C18:1) (r = - 0.79, p < 0.000) as well as lower proportion of palmitic acid (C16:0) (r = 0.51, p = 0.037), making varieties with this genotype (i.e. Zarrazi and Tounsi) producing more monounsaturated oleic acid (C18: 1) than saturated acid. These varieties could be thus used as nutraceuticals and functional food. CONCLUSION: The SAD.1 association with the oleic acid composition of olive oil was identified among the studied varieties. This correlation fluctuated between studied varieties, which might elucidate variability in lipidic composition among them and therefore reflecting genetic diversity through differences in gene expression and biochemical pathways. SAD locus would represent an excellent marker for identifying interesting amongst virgin olive oil lipidic composition.

Proteomic Analysis of the Chlorophyta Dunaliella New Strain AL-1 Revealed Global Changes of Metabolism during High Carotenoid Production.

The green microalgae Dunaliella genus is known for the production of high added value molecules. In this study, strain AL-1 was isolated from the Sebkha of Sidi El Hani (Sousse, Tunisia). This isolate was identified both morphologically and genetically via 18S rRNA gene sequence as a member of the genus Dunaliella. Strain AL-1 was found to be closely related to Dunaliella salina, Dunaliella quartolecta and Dunaliella polymorpha with more than 97% similarity. Response surface methodology was used to maximize carotenoid production by strain AL-1 by optimizing its growth conditions. The highest carotenoid content was obtained at salinity: 51, light intensity: 189.89 µmol photons·m-2·s-1, and nitrogen: 60 mg·L-1. Proteomic profiling, using two-dimensional gel electrophoresis, was performed from standard and optimized cultures. We detected 127 protein spots which were significantly differentially expressed between standard and optimized cultures. Among them 16 protein spots were identified with mass spectrometry and grouped into different functional categories using KEGG (Kyoto Encyclopedia of Genes and Genomes) such as photosynthetic Calvin cycle, regulation/defense, energy metabolism, glycolysis, and cellular processes. The current study could be of great interest in providing information on the effect of stressful conditions in microalgae carotenoid production.

Tetraselmis species for environmental sustainability: biology, water bioremediation, and biofuel production.

With increasing demand of fossil fuels and water pollution and their environmental impacts, marine green microalgae have gained special attention in both scientific and industrial  fields. This is due to their fast growth in non-arable lands with high photosynthetic activity, their metabolic plasticity, as well as their high CO2 capture capacity. Tetraselmis species, green and eukaryotic microalgae, are not only considered as a valuable source of biomolecules including pigments, lipids, and starch but also widely used in biotechnological applications. Tetraselmis cultivation for high-value biomolecules and industrial use was demonstrated to be a non-cost-effective strategy because of its low demand in nutrients, such as phosphorus and nitrogen. Recently, phycoremediation of wastewater rich in nutrients, chemicals, and heavy metals has become an efficient and economic-alternative that allows the detoxification of waters and induces mechanisms in algal cells for biomolecules rich-energy synthesis to regulate their metabolic pathways. This review aims to shed light on Tetraselmis species for their different culture conditions and metabolites bioaccumulation, as well as their human health and environmental applications. Additionally, phycoremediation of contaminants associated to biofuel production in Tetraselmis cells and their different intracellular and extracellular mechanisms have also been investigated.

Improving the shelf life of minced beef by Cystoseira compressa polysaccharide during storage.

A polysaccharide extracted from the brown alga Cystoseira compressa (CCPS) was evaluated as a food additive to extend the shelf-life of raw beef meat. The antioxidant potential of CCPS was demonstrated by its inhibition of ß-carotene bleaching (64.28 %), superoxide radicals (70.12 %), and hydroxyl radicals (93 %) at a concentration of 10 mg/ml. The polysaccharide also showed antibacterial activity with MIC values between 6.25 mg/ml and 50 mg/ml against five foodborne pathogenic bacteria. Furthermore, CCPS exhibited excellent functional, foaming, and emulsifying properties. Furthermore, microbiological and chemical effects of CCPS at concentrations equivalent to 1 MIC (CCPS-1), 2 MIC (CCPS-2), and 4 MIC (CCPS-3) were conducted. Chemical analyses showed that treated beef had significantly reduced TBARS levels below 2 mg MDA/kg at day 14. The treatment also decreased carbonyl groups, improved heme iron transformation, inhibited microbial growth (p < 0.05), and kept MetMb levels below 40 % by day 14. Moreover, two multivariate approaches, principal component analysis (PCA) and hierarchical cluster analysis (HCA), were effectively used to analyze the results characterizing the main attributes of the stored meat samples. In conclusion, these findings demonstrated that CCPS could be employed as a functional and bioactive component in the meat industry.

Enhanced growth and metabolite production from a novel strain of Porphyridium sp.

Microalgae are capable of generating numerous metabolites that possess notable biological activities and hold substantial promise for various industrial applications. Nevertheless, the taxonomic diversity of these photosynthetic microorganisms has not received thorough investigation. Using the 18S rRNA encoding gene, a recently discovered strain originating from the Tunisian coast (the governorate of Mahdia) was identified as a member of the Porphyridium genus. The growth response as well as the metabolite accumulation of Porphyridium sp. to different culture media (Pm, F/2, and Hemerick) was investigated over a period of 52 days. The highest biomass production was recorded with Pm medium (2 × 107 cell/mL). The apparent growth rates (µ) and the doubling time (Dt) were about 0.081 day-1 and 12.34 days, respectively. The highest chlorophyll a (0.678 ± 0.005 pg/cell), total carotenoids (0.18 ± 0.003 pg/cell), phycoerythrin (3.88 ± 0.003 pg/cell), and proteins (14.58 ± 0.35 pg/cell) contents were observed with F/2 medium. Cultivating Porphyridium sp. in both F/2 and Hemerick media yielded similar levels of starch accumulation. The Hemerick medium has proven to be the most suitable for the production of lipids (2.23% DW) and exopolysaccharides (5.41 ± 0.56 pg/cell).

Identification of critical residues for the activity and thermostability of Streptomyces sp. SK glucose isomerase.

The role of residue 219 in the physicochemical properties of D-glucose isomerase from Streptomyces sp. SK strain (SKGI) was investigated by site-directed mutagenesis and structural studies. Mutants G219A, G219N, and G219F were generated and characterized. Comparative studies of their physicochemical properties with those of the wild-type enzyme highlighted that mutant G219A displayed increased specific activity and thermal stability compared to that of the wild-type enzyme, while for G219N and G219F, these properties were considerably decreased. A double mutant, SKGI F53L/G219A, displayed a higher optimal temperature and a higher catalytic efficiency than both the G219A mutant and the wild-type enzyme and showed a half-life time of about 150 min at 85 °C as compared to 50 min for wild-type SKGI. Crystal structures of SKGI wild-type and G219A enzymes were solved to 1.73 and 2.15 Å, respectively, and showed that the polypeptide chain folds into two structural domains. The larger domain consists of a (ß/α)8 unit, and the smaller domain forms a loop of α helices. Detailed analyses of the three-dimensional structures highlighted minor but important changes in the active site region as compared to that of the wild-type enzyme leading to a displacement of both metal ions, and in particular that in site M2. The structural analyses moreover revealed how the substitution of G219 by an alanine plays a crucial role in improving the thermostability of the mutant enzyme.

Changes in the catalytic properties and substrate specificity of Bacillus sp. US149 maltogenic amylase by mutagenesis of residue 46.

Maltogenic amylase from Bacillus sp. US149 (MAUS149) is a cyclodextrin (CD)-degrading enzyme with a high preference for CDs over maltooligosaccharides. In this study, we investigated the roles of residue Asp46 in the specificity and catalytic properties of MAUS149 by using site-directed mutagenesis. Three mutated enzymes (D46V, D46G and D46N) were constructed and studied. The three mutants were found to be similar to the wild-type MAUS149 regarding thermoactivity, thermostability and pH profile. Nevertheless, the kinetic parameters for all the substrates of the mutant enzymes D46V and D46G were altered enormously as compared with those of the wild type. Indeed, the K(m) values of MAUS149/D46G for all substrates were strongly increased. Nevertheless, the affinity and catalytic efficiency of MAUS149/D46V toward ß-CD were increased fivefold as compared with those of MAUS149. Molecular modelling suggests that residue D46 forms a salt bridge with residue K282. This bond would maintain the arrangement of side chains of residues Y45 and W47 in a particular orientation that promotes access to the catalytic site and maintains the substrate therein. Hence, any replacement with uncharged amino acids influenced the flexibility of the gate wall at the substrate binding cleft resulting in changes in substrate selectivity.

Expression of a Copper Activated Xylanase in Yeast: Location of the His-Tag in the Protein Significantly Affects the Enzymatic Properties.

A copper activated xylanase produced by E. coli BL21 was expressed in Pichia pastoris using the pGAPZαB expression vector. Two recombinant GH11 xylanase forms were obtained (N-His-rXAn11 and N-C-His-rXAn11). The findings revealed that the two recombinant xylanases displayed different behaviors toward the copper. In the presence of 3-mM Cu2+, the relative activity of the N-His-rXAn11 was enhanced by about 52%. However, the xylanase activity of the N- and C-terminal tagged one (N-C-His-rXAn11) was strongly inhibited by copper. In the presence of 3-mM Cu2+, the N-His-rXAn11 revealed to be thermostable at 60 °C with a half-life of 10 min. However, the N-C-His-rXAn11 was noted to be unstable since it was inactivated after 15 min of incubation at 55 °C. 3D models of the two recombinant forms showed that the created copper site in the N-His-rXAn11 was loosed in the C-terminal tagged protein. The C-terminal tag could trigger some structural changes with a notable displacement of secondary structures leading to great hindrance of the active site due to high fluctuations and probably new interactions among the N- and C-terminal amino acids.

Apigenin analogues as SARS-CoV-2 main protease inhibitors: In-silico screening approach.

The COVID-19 new variants spread rapidly all over the world, and until now scientists strive to find virus-specific antivirals for its treatment. The main protease of SARS-CoV-2 (Mpro) exhibits high structural and sequence homology to main protease of SARS-CoV (93.23% sequence identity), and their sequence alignment indicated 12 mutated/variant residues. The sequence alignment of SARS-CoV-2 main protease led to identification of only one mutated/variant residue with no significant role in its enzymatic process. Therefore, Mpro was considered as a high-profile drug target in anti-SARS-CoV-2 drug discovery. Apigenin analogues to COVID-19 main protease binding were evaluated. The detailed interactions between the analogues of Apigenin and SARS-CoV-2 Mpro inhibitors were determined as hydrogen bonds, electronic bonds and hydrophobic interactions. The binding energies obtained from the molecular docking of Mpro with Boceprevir, Apigenin, Apigenin 7-glucoside-4'-p-coumarate, Apigenin 7-glucoside-4'-trans-caffeate and Apigenin 7-O-beta-d-glucoside (Cosmosiin) were found to be -6.6, -7.2, -8.8, -8.7 and -8.0 kcal/mol, respectively. Pharmacokinetic parameters and toxicological characteristics obtained by computational techniques and Virtual ADME studies of the Apigenin analogues confirmed that the Apigenin 7-glucoside-4'-p-coumarate is the best candidate for SARS-CoV-2 Mpro inhibition.

Ginger: a systematic review of clinical trials and recent advances in encapsulation of its bioactive compounds.

Recently, the numbers of studies on natural products have considerably increased owing to their exceptional biological activities and health benefits. Their pharmacological attributes have played an immense role in detecting natural and safe alternative therapeutics, consequently extending their industrial applications. In this line, ginger (Zingiber officinale) has been gaining wide attention owing to its bioactive compounds, such as phenolic and terpene compounds. Ginger has a great pharmacological and biological potential in the prevention and treatment of various diseases, namely colds, nausea, arthritis, migraines and hypertension. However, these bioactive compounds are unstable and susceptible to degradation, volatilization and oxidation during extraction and processing, mainly owing to their exposure to environments with adverse conditions, such as high temperature, the presence of O2 and light. In this sense, this current review covers a wide range of topics, starting from the chemical profile and biological properties of ginger bioactive compounds (GBCs), their clinical effectiveness for the treatment of diseases and the application of different encapsulation methods (molecular inclusion, spray drying, complex coacervation, ionic strength and nanoemulsions) to protect and improve their application in food products. This work summarizes the fundamental principles of, recent progress in and effectiveness of different methods regarding the physicochemical, structural and functional properties of encapsulated GBCs. The potential use of encapsulated GBCs as a promising active ingredient to be applied in different food products is discussed in detail.

Conception of an environmental friendly O/W cosmetic emulsion from microalgae.

The development of eco-friendly cosmetic such as those from microalgae for skin regeneration and collagen synthesis has gained a great interest worldwide. Accordingly, the potential of microalgae biomass as source of anti-aging cosmetic cream with high antioxidant activity has been investigated. Stabilities and sensory characteristics of cosmetic creams supplemented with Spirulina, Tetraselmis sp. and Dunaliella sp. at 0.5, 1.5 and 2.5%, respectively, revealed a conservation of physico-chemical and preliminary stability properties of formulations. To analyze physico-chemical and textural parameters, accelerated stability study was evaluated under two thermal conditions (25 and 40 °C) during 90 days. Results showed that pH values of all formulations were within the limits of normal skin pH range under storage time at 25 and 40 °C. During this period, the colored creams showed a significant changes of a* and b* indices, reflecting the instability of microalgae colors. Microalgae modified the textural characteristics of emulsions. The Tetraselmis sp. containing-cream had the lowest (P < 0.05) values of hardness, springiness, and cohesiveness. The 0.5% Spirulina containing-cream had the best stable consistency and adhesiveness under time and temperature variations. It exhibited the best properties to be used for skin care products. Thanks to their high content in bioactive macromolecules, microalgae considerably improved the antioxidant activity of the new formulated skin creams.

Effect of heavy metals mixture on the growth and physiology of Tetraselmis sp.: Applications to lipid production and bioremediation.

Phycoremediation of heavy metals from contaminated waters by oleaginous microalgae is an eco-friendly and emerging trend. Different concentrations of toxic metals such as nickel (Ni), chromium (Cr) and cobalt (Co) were added in Tetraselmis sp. culture media. Mixture Design was used to model the effect of these metals on cell growth, lipid production and heavy metals removal. Tetraselmis sp. was identified as an outstanding Ni, Cr and Co accumulator with bioconcentration factors of 675.17, 584.9 and 169.81 within binary mixtures (Ni × Cr), (Cr × Co) and (Cr × Co) at 6 mg/L, respectively. Optimization studies showed that the highest cell growth (9.22 × 105 cells/mL), lipid content (31% Dry Weight) and metals removal (91%) were obtained with the optimum binary mixture Ni (54.45 %) and Cr (45.45 %). This work presents interesting results revealing the potential of Tetraselmis sp. for nickel removal up to 97 % combined to its potential for biodiesel production.

Microalgae as feedstock for bioactive polysaccharides.

Due to the increase in industrial demand for new biosourced molecules (notably bioactive exopolysaccharides (EPS)), microalgae are gaining popularity because of their nutraceutical potential and benefits health. Such health effects are delivered by specific secondary metabolites, e.g., pigments, exopolysaccharides, polyunsaturated fatty acids, proteins, and glycolipids. These are suitable for the subsequent uses in cosmetic, nutraceutical, pharmaceutical, biofuels, biological waste treatment, animal feed and food fields. In this regard, a special focus has been given in this review to describe the various methods used for extraction and purification of polysaccharides. The second part of the review provides an up-to-date and comprehensive summary of parameters affecting the microalgae growth and insights to maximize the metabolic output by understanding the intricacies of algal development and polysaccharides production. In the ultimate part, the health and nutraceutical claims associated with marine algal bioactive polysaccharides, explaining their noticeable potential for biotechnological applications, are summarized and comprehensively discussed.

SNP discovery and structural insights into OeFAD2 unravelling high oleic/linoleic ratio in olive oil.

Fatty Acid Desaturase 2 (FAD2), a key enzyme in the fatty acid biosynthesis pathway, is involved in the desaturation and conversion of oleic acid to linoleic acid. Therefore, it plays a crucial role in oleic/linoleic acid ratio and the quality of olive oil. DNA sequencing of 19 FAD2 genes from a set of olive oil varieties revealed several single-nucleotide polymorphisms (SNPs) and highlighted associations between some of the SNPs and saturated fatty acids contents. This was further confirmed by SNP-interaction and machine learning approach. Haplotype diversity analysis led to the discovery of three highly polymorphic SNPs and four haplotypes harboring differential oleic/linoleic acid ratios. Moreover, a combination of molecular modeling and docking experiments allowed a deeper and better understanding of the structure-function relationship of the FAD2 enzyme. Sequence patterns and variations involved in the regulation of the FAD2 activity were also identified. Furthermore, S82C and H213N substitutions in OeFAD2 make the Oueslati variety more interesting in terms of fatty acid profile and oleic acid level.

In silico evidence of antiviral activity against SARS-CoV-2 main protease of oligosaccharides from Porphyridium sp.

The coronavirus pandemic (COVID-19) has created an urgent need to develop effective strategies for prevention and treatment. In this context, therapies against protease Mpro, a conserved viral target, would be essential to contain the spread of the virus and reduce mortality. Using combined techniques of structure modelling, in silico docking and pharmacokinetics prediction, many compounds from algae were tested for their ability to inhibit the SARS-CoV-2 main protease and compared to the recent recognized drug Paxlovid. The screening of 27 algal molecules including 15 oligosaccharides derived from sulfated and non-sulphated polysaccharides, eight pigments and four poly unsaturated fatty acids showed high affinities to interact with the protein active site. Best candidates showing high docking scores in comparison with the reference molecule were sulfated tri-, tetra- and penta-saccharides from Porphyridium sp. exopolysaccharides (SEP). Structural and energetic analyses over 100 ns MD simulation demonstrated high SEP fragments-Mpro complex stability. Pharmacokinetics predictions revealed the prospects of the identified molecules as potential drug candidates.

Pistachio Hull Extract as a Practical Strategy to Extend the Shelf Life of Raw Minced Beef: Chemometrics in Quality Evaluation.

The agricultural processing industry produces a notable quantity of by-products rich in bioactive compounds, which can be exploited for agri-food applications. From pistachio industrial processing, pistachio's hull is one of the major by-products. This work aimed to evaluate the potential of pistachio hull, as a potential source of natural antioxidant, to preserve the meat quality. Here, we investigated the impact of aqueous pistachio hull extract (PHE) at 0.156% (PHE1), 0.312% (PHE2), and 0.625% (PHE3) on the quality of raw minced beef meat stored for 14 days at 4°C. At the end of storage, mesophilic total viable plate, psychotropic and Enterobacteriaceae counts, showed significantly lower (P < 0.05) microbial count in PHE samples. PHE3 revealed a powerful inhibitory effect on lipid/protein oxidation, and sensory characteristics were positively (P < 0.05) affected. Principal component analysis and heat map indicated complex and close synchronized relations among lipid/protein oxidation processes, microbial loads, and sensory attributes. Obtained results using univariate and multivariate statistical analysis underlined the importance of using different mathematical approaches, which are complementary to each other and could provide considerable information about the minced beef meat treated by PHE. Therefore, compared to synthetic antioxidants, PHE could be a clean-label alternative that can protect and enhance the quality of meat products.