Characterization of cellular toxicity induced by sub-lethal inorganic mercury in the marine microalgae Chlorococcum dorsiventrale isolated from a metal-polluted coastal site

Mercury (Hg) is a global pollutant that affects numerous marine aquatic ecosystems. We isolated Chlorococcum dorsiventrale Ch-UB5 microalga from coastal areas of Tunisia suffering from metal pollution and analyzed its tolerance to Hg. This strain accumulated substantial amounts of Hg and was able to remove up to 95% of added metal after 24 and 72 h in axenic cultures. Mercury led to lesser biomass growth, higher cell aggregation, significant inhibition of photochemical activity, and appearance of oxidative stress and altered redox enzymatic activities, with proliferation of starch granules and neutral lipids vesicles. Such changes matched the biomolecular profile observed using Fourier Transformed Infrared spectroscopy, with remarkable spectral changes corresponding to lipids, proteins and carbohydrates. C. dorsiventrale accumulated the chloroplastic heat shock protein HSP70B and the autophagy-related ATG8 protein, probably to counteract the toxic effects of Hg. However, long-term treatments (72 h) usually resulted in poorer physiological and metabolic responses, associated with acute stress. C. dorsiventrale has potential use for Hg phycoremediation in marine ecosystems, with the ability to accumulating energetic reserves that could be used for biofuel production, supporting the notion of using of C. dorsiventrale for sustainable green chemistry in parallel to metal removalThis work was supported by grant from the Tunisian Ministry of Higher Education and Scientific Research. This work was also funded by the Spanish Ministry of Science and Technology (AEI) (projects AGL2014-53771-R and AGL2017-87591-R

Antidiabetic potential of mucilage fraction extracted from Astragalus gyzensis seeds

The objective of the current work is to extract a new mucilage fraction from Astragalus gyzensis Bunge. seeds, which are collected from the El-Oued province (septentrional Algerian Sahara) and evaluated for their antidiabetic potential. The mucilage fraction is obtained using hot water extraction followed by alcoholic precipitation of polysaccharides by cold ethanol (96%). The primary investigation was performed by describing the main structural features of the extract through colorimetric assays, Fourier-transform infrared spectroscopy and thin-layer chromatography analysis using two systems. Biological activity was also monitored by antidiabetic activity by testing the inhibition of α-amylase and α-glucosidase enzymes in vitro. The extraction yield was 20.69%. The chemical composition mainly consisted of 78.60±0.29% carbohydrates, among them 63.92±0.67% neutral sugar, 15.78±0.76% uronic acid, 8.08±0.04% proteins and 2.57±0.05% phenolic compounds. The results obtained by thin-layer chromatography analysis showed the dominance of mannose and galactose. Fourier-transform infrared spectrum showed characteristic bands expected galactomannans. The investigations highlighted the antihyperglycemic effect in a dose-dependent manner by the inhibition of the α-amylase enzyme (IC50=0.8±0.005 mg/mL). These factors make it suitable for the industrial application of dietary supplement fiber made for diabetic individuals. DOI: http://dx.doi.org/10.5281/zenodo.761853

Cellulosomes de Clostridium cellulolyticum (diversité, synergies et régulation)

Les cellulosomes de Clostridium cellulolyticum sont très hétérogènes du point de vue de leur composition en enzymes. Chaque cellulosome contient au maximum huit enzymes alors que 36 protéines à dockérine ont été identifiées dans les cellulosomes produits d'une culture sur cellulose. Plusieurs combinaisons enzymatiques sont à priori possibles dans les cellulosomes. Mon travail de thèse a consisté d'une part à étudier la diversité des cellulosomes produits et les synergies entre complexes. De ce fait, différentes fractions de cellulosomes issues d'une chromatographie d'échange d'ions montrent à la fois la présence de protéines enzymatiques communes à toutes les fractions, et la présence de protéines particulières à certaines fractions. Des composants enzymatiques non caractérisés ont été identifiés dans certaines fractions actives sur des polymères particuliers (par exemple, 3 xylanases dans la fraction la plus active sur xylane). Des synergies entre les cellulosomes de différentes fractions ont été mises en évidence. Cette coopération entre cellulosomes apporterait au mélange naturel une très grande efficacité dans la dégradation des polymères végétaux. En deuxième lieu, mon travail a consisté à étudier la régulation de l'expression des gènes du système cellulolytique. Un opéron cip-cel codant pour les enzymes clés des cellulosomes (facteur d'assemblage cipC ainsi que les cellulases majoritaires Cel48F et Ce19E), est régulé par répression catabolique ainsi que par un régulateur transcriptionnel spécifique appartenant à la famille LysR (appelé GlyR1). En plus de l'opéron, on trouve d'autres gènes situés ailleurs (appelés gènes isolés) codant pour d'autres composants du système. Parmi ces gènes, cel5A et cel5I sont également soumis à la répression catabolique, alors que le gène cel440 est régulé par activation catabolique. Le gène cel5D est régulé par un régulateur transcriptionnel spécifique appartenant à la famille AraC, nommé GlyR2. Les différents régulateurs identifiés montrent la grande complexité du phénomène de la régulation des gènes de C. cellulolyticum.AIX-MARSEILLE1-BU Sci.St Charles (130552104) / SudocSudocFranceF

Optimization of lipids’ ultrasonic extraction and production from Chlorella sp. using response-surface methodology

Abstract Background Three steps are very important in order to produce microalgal lipids: (1) controlling microalgae cultivation via experimental and modeling investigations, (2) optimizing culture conditions to maximize lipids production and to determine the fatty acid profile the most appropriate for biodiesel synthesis, and (3) optimizing the extraction of the lipids accumulated in the microalgal cells. Methods Firstly, three kinetics models, namely logistic, logistic-with-lag and modified Gompertz, were tested to fit the experimental kinetics of the Chlorella sp. microalga culture established on standard conditions. Secondly, the response-surface methodology was used for two optimizations in this study. The first optimization was established for lipids production from Chlorella sp. culture under different culture conditions. In fact, different levels of nitrate concentrations, salinities and light intensities were applied to the culture medium in order to study their influences on lipids production and determine their fatty acid profile. The second optimization was concerned with the lipids extraction factors: ultrasonic’s time and temperature, and chloroform-methanol solvent ratio. Results All models (logistic, logistic-with-lag and modified Gompertz) applied for the experimental kinetics of Chlorella sp. show a very interesting fitting quality. The logistic model was chosen to describe the Chlorella sp. kinetics, since it yielded the most important statistical criteria: coefficient of determination of the order of 94.36%; adjusted coefficient of determination equal to 93.79% and root mean square error reaching 3.685 cells · ml− 1. Nitrate concentration and the two interactions involving the light intensity (Nitrate concentration × light intensity, and salinities × light intensity) showed a very significant influence on lipids production in the first optimization (p < 0.05). Yet, only the quadratic term of chloroform-methanol solvent ratio showed a significant influence on lipids extraction relative to the second step of optimization (p < 0.05). The two most abundant fatty acid methyl esters (≈72%) derived from the Chlorella sp. microalga cultured in the determined optimal conditions are: palmitic acid (C16:0) and oleic acid (C18:1) with the corresponding yields of 51.69% and 20.55% of total fatty acids, respectively. Conclusions Only the nitrate deficiency and the high intensity of light can influence the microalgal lipids production. The corresponding fatty acid methyl esters composition is very suitable for biodiesel production. Lipids extraction is efficient only over long periods of time when using a solvent with a 2/1 chloroform/methanol ratio

Influence of Zeolite on fatty acid composition and egg quality in Tunisian Laying Hens

<p>Abstract</p> <p>Background</p> <p>The health benefits of omega-3 and omega-6 polyunsaturated fatty acids (PUFA) are generally recognized. Unfortunately, in most Mediterranean countries, the recommended daily intake of these compounds is rarely met. Therefore, enrichment of commonly occurring foods can boost intake of these fatty acids. In this regard, eggs are an interesting target, as they form an integral part of the diet.</p> <p>Result</p> <p>Zeolite (Clinoptilolites) was added to Laying Hens feed at concentrations 1% or 2% and was evaluated for its effects on performance of the production and on egg quality. The Laying Hens were given access to 110 g of feed mixtures daily that was either a basal diet or a ‘zeolite diet’ (the basal diet supplemented with clinoptilolite at a level of 1% or 2%). It was found that zeolite treatment had a positive and significatif (p < 0.05) effect on some parameters that were measured like egg height and eggshell strength. While dietary zeolite supplementation tended to/or has no significant effects on total egg, eggshell, yolk and albumen weights. It was found also that zeolite mainly increases level of polyunsaturated fatty acids in egg.</p> <p>Conclusion</p> <p>This study showed the significance of using zeolite, as a feed additive for Laying Hens, as part of a comprehensive program to control egg quality and to increase level of polyunsaturated fatty acids on egg.</p

Production and purification of fucoxanthins and β-carotenes from Halopteris scoparia and their effects on digestive enzymes and harmful bacteria

International audienc

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

International audienc

Deciphering the Biological Activities of Dunaliella sp. Aqueous Extract from Stressed Conditions on Breast Cancer: from in Vitro to in Vivo Investigations

International audienceIn order to harness local resources to improve well-being and human health, we aim in this study to investigate if the microalgae Dunaliella sp. isolated from the Tunisian coastal zone possesses any anticancer activity. Dunaliella sp. was cultured under normal (DSC) or stressed (DSS) conditions and extracted using different procedures. The biological activity assessment was performed on the Triple Negative Breast Cancer (TNBC) using 4T1 murine cells as a model. Results indicate that: (i) aqueous extract was the most cytotoxic compared to ethanolic and hydroalcoholic extracts; (ii) DSS activity was superior to that of DSC. DSS extracts induced apoptosis rather than necrosis, as evidenced by DNA fragmentation, PARP-1 cleavage and caspase-3 activation. Evaluation in an orthotopic TNBC model validated the anticancer activity in vivo. Intratumoral injection of DSS extract resulted in reduced tumor growth and an enhanced immune system activation. On the transcriptional side, the expression level of the immunosuppressive enzyme Arg-1 was decreased, as well as those of NOS-2 and COX-2 genes. These results suggest a potential anticancer activity of Tunisian Dunaliella sp. deserving further attention

Identification of a new oat β-amylase by functional proteomics

International audienceOat (Avena sativa L.) seed extracts exhibited a high degree of catalytic activity including amylase activities. Proteins in the oat seed extracts were optimized for their amylolytic activities. Oat extract with amylolytic activity was separated by SDS-PAGE and a major protein band with an apparent molecular mass of 53 kDa was subjected to tryptic digestion. The generated amino acid sequences were analyzed by liquid chromatography–tandem mass spectrometry (LC/ESI/MS/MS) and database searches. These sequences were used to identify a partial cDNA from expressed sequence tags (ESTs) of A. sativa L. Based upon EST sequences, a predicted full-length gene was identified, with an open reading frame of 1464 bp encoding a protein of 488 amino acid residues (AsBAMY), with a theoretical molecular mass of 55 kDa identified as a β-amylase belonging to the plant β-amylase family. Primary structure of oat β-amylase (AsBAMY) protein indicated high similarity with other β-amylase from other cereals such as wheat (Triticum aestivum), barley (Hordeum vulgare), and rye (Secale cereale) with two conserved Glu residues (E184 and E378) assigned as the “putative” catalytic residues which would act as an acid and base pair in the catalytic process. In addition, a 3D-model of AsBAMY was built from known X-ray structures and sequence alignments. A similar core (β/α)8-barrel architecture was found in AsBAMY like the other cereal β-amylases with a specific location of the active site in a pocket-like cavity structure made at one end of this core (β/α)8-barrel domain suggesting an accessibility of the non-reducing end of the substrate and thus confirming the results of AsBAMY exo-acting hydrolase