Taxonomy, purification and chemical characterization of four bioactive compounds from new Streptomyces sp. TN256 strain

A new actinomycete strain designated TN256, producing antimicrobial activity against pathogenic bacteria and fungi, was isolated from a Tunisian Saharan soil. Morphological and chemical studies indicated that strain TN256 belonged to the genus Streptomyces. Analysis of the 16S rDNA sequence of strain TN256 showed a similarity level ranging between 99.79 and 97.8% within Streptomyces microflavus DSM 40331T and Streptomyces griseorubiginosus DSM 40469T respectively. The comparison of its physiological characteristics showed significant differences with the nearest species. Combined analysis of the 16 S rRNA gene sequences (FN687758), fatty acids profile, and results of physiological and biochemical tests indicated that there were genotypic and phenotypic differentiations of that isolate from other Streptomyces species neighbours. These date strongly suggest that strain TN256 represents a novel species with the type strain Streptomyces TN256 (=CTM50228T). Experimental validation by DNA–DNA hybridization would be required for conclusive confirmation. Four active products (1–4) were isolated from the culture broth of Streptomyces TN256 using various separation and purification steps and procedures. 1: N-[2-(1H-indol-3-yl)-2 oxo-ethyl] acetamide ‘alkaloid’ derivative; 2: di-(2-ethylhexyl) phthalate, a phthalate derivative; 3: 1-Nonadecene and 4: Cyclo (l-Pro-l-Tyr) a diketopiperazine ‘DKP’ derivative. The chemical structure of these four active compounds was established on the basis of spectroscopic studies NMR and by comparing with data from the literature. According to our biological studies, we showed in this work that the pure compounds (1–4) possess antibacterial and antifungal activities

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

Inhibition of Fungi and Gram-Negative Bacteria by Bacteriocin BacTN635 Produced by Lactobacillus plantarum sp. TN635

The aim of this study was to evaluate 54 lactic acid bacteria (LAB) strains isolated from meat, fermented vegetables and dairy products for their capacity to produce antimicrobial activities against several bacteria and fungi. The strain designed TN635 has been selected for advanced studies. The supernatant culture of this strain inhibits the growth of all tested pathogenic including the four Gram-negative bacteria (Salmonella enterica ATCC43972, Pseudomonas aeruginosa ATCC 49189, Hafnia sp. and Serratia sp.) and the pathogenic fungus Candida tropicalis R2 CIP203. Based on the nucleotide sequence of the 16S rRNA gene of the strain TN635 (1,540 pb accession no FN252881) and the phylogenetic analysis, we propose the assignment of our new isolate bacterium as Lactobacillus plantarum sp. TN635 strain. Its antimicrobial compound was determined as a proteinaceous substance, stable to heat and to treatment with surfactants and organic solvents. Highest antimicrobial activity was found between pH 3 and 11 with an optimum at pH = 7. The BacTN635 was purified to homogeneity by a four-step protocol involving ammonium sulfate precipitation, centrifugal microconcentrators with a 10-kDa membrane cutoff, gel filtration Sephadex G-25, and C18 reverse-phase HPLC. SDS-PAGE analysis of the purified BacTN635, revealed a single band with an estimated molecular mass of approximately 4 kDa. The maximum bacteriocin production (5,000 AU/ml) was recorded after a 16-h incubation in Man, Rogosa, and Sharpe (MRS) medium at 30 °C. The mode of action of the partial purified BacTN635 was identified as bactericidal against Listeria ivanovii BUG 496 and as fungistatic against C. tropicalis R2 CIP203

Bioactive Secondary Metabolites from a New Terrestrial Streptomyces sp. TN262

During our search for Streptomyces spp. as new producers of bioactive secondary metabolites, the ethyl acetate extract of the new terrestrial Streptomyces isolate TN262 delivered eight antimicrobially active compounds. They were identified as 1-acetyl-β-carboline (1), tryptophol (2), cineromycin B (3), 2,3-dihydrocineromycin B (4), cyclo-(tyrosylprolyl) (5), 3-(hydroxyacetyl)-indole (6), brevianamide F (7), and cis-cyclo-(l-prolyl-l-leucyl) (8). Three further metabolites were detected in the unpolar fractions using GC–MS and tentatively assigned as benzophenone (9), N-butyl-benzenesulfonamide (10), and hexanedioic acid-bis-(2-ethylhexyl) ester (11). This last compound is known as plasticizer derivatives, but it has never been described from natural sources. In this article, we describe the identification of the new Streptomyces sp. isolate TN262 using its cultural characteristics, the nucleotide sequence of the corresponding 16S rRNA gene and the phylogenetic analysis, followed by optimization, large-scale fermentation, isolation of the bioactive constituents, and determination of their structures. The biological activity of compounds (2), (3), (4), and those of the unpolar fractions was addressed as well

Bis[tris(propane-1,3-diamine-κ2N,N′)nickel(II)] diaquabis(propane-1,3-diamine-κ2N,N′)nickel(II) hexabromide dihydrate

In the title compound, [Ni(C3H10N2)3]2[Ni(C3H10N2)2(H2O)2]Br6·2H2O, one Ni2+ cation, located on an inversion centre, is coordinated by four N atoms from two ligands and by two water O atoms. The other Ni2+ cation, located in a general position, is coordinated by six N atoms from three ligands. In both cases, the Ni2+ cation has an octahedral coordination environment. The overall structural cohesion is ensured by three types of hydrogen bonds, N—H...Br, O—H...Br and O—H...O, which connect the two types of complex cations, the bromide counter-anions and the lattice water molecules into a three-dimensional network

Cry1Ac toxicity enhancement towards lepidopteran pest Ephestia kuehniella through its protection against excessive proteolysis

Bacillus thuringiensis has been extensively used in agroecosystems for four decades due to its high specific toxicity. Strategies based on B. thuringiensis proteins combinations for the improvement of its activity present an important focus for biopesticides development. However, the widespread use of B. thuringiensis δ-endotoxins has often been challenged by a lack of understanding of the target insect physiology as well as its midgut biochemistry.In the present investigation, we have evidenced and explained the toxicity improvement of Cry1Ac δ-endotoxins against Ephestia kuehniella larvae through in vivo combination with P20 helper protein. Tracking the fate of Cry1Ac in tested midgut larvae showed considerable differences between δ-endotoxins produced in the presence of P20 and those produced in its absence which could explain the obtained larvicidal activity enhancement. The P20 presence slightly increased Cry1Ac inclusions solubility in E. kuehniella midgut conditions. However, a protection against excessive degradation of protoxin and toxin forms of Cry1Ac was strongly decreased in the case of δ-endotoxins produced in the presence of P20 as compared to those from P20 lacking control. Thus, the P20 protective effect on Cry1Ac after larvae ingestion has been proven. This finding could be helpful to further understand the roles of P20 helper protein in toxicity enhancement of B. thuringiensis toxins.This work was supported by grants from Ministry of Higher Education and Scientific Research

Synthesis and characterization of a hybrid material (C10H28N4) [CoCl4]2 using Hirshfeld surface, vibrational and optical spectroscopy, DFT calculations and biological activities

International audienceThe preparation and structural characterization of a new complex compound (C10H28N4) [CoCl4]2 (abbreviated BAPCO) were reported. The crystal was grown by the room temperature slow evaporation method and characterized by single crystal X-Ray diffraction. The functional groups present in BAPCO were characterized by several techniques such as FT-IR, UV-Vis absorption and photoluminescence spectroscopy, thermal measurement, Hirshfeld surface analysis and DFT investigation. The title compound crystallizes in the P21/n space group of the monoclinic system with the following cell parameters: a = 18.5765(11) Å, b = 7.1390(4) Å, c = 18.6346(11) Å, β = 110.858(3)°, Z = 4 and V = 2309.3(2) Å3. The structure consists of an alternation along the a-axis of organic layers formed by [C10H28N4]4+cations and inorganic layers built up of isolated tetrahedral [CoCl4]2−. The crystal cohesion is ensured by a network of Nsingle bondH … Cl and Csingle bondH … Cl hydrogen bonds. Vibrational and optical properties were explored by means of experimental techniques along with DFT and TDDFT calculations. Furthermore, frontier molecular orbital analysis (HOMO-LUMO) was accomplished to understand the chemical stability of BAPCO, and the activation of thermodynamic parameters are calculated. Good agreement was found between theoretical and experimental results. The bioassay results showed that the structure exhibits significant antibacterial activity

Genome sequence analysis of a novel Bacillus thuringiensis strain BLB406 active against Aedes aegypti larvae, a novel potential bioinsecticide.

International audienceBLB406 is a novel isolate of Bacillus thuringiensis with a larvicidal activity against Aedes aegypti larvae. It displays original plasmidic and crystal protein patterns. The present work reported molecular and bioinformatic analyses for the genome sequence of BLB406 using MiSeq Illumina next-generation sequencing technology. The reads were assembled by Velvet tool. Using RAST program and PGAAP, the genome of BLB406 strain was shown to contain 6297 genes corresponding to 5924 protein coding sequences. The BLB406 genome investigation with BtToxin_scanner program shows that this strain has an original and different combination of toxins compared to the published ones: five cry genes (cry11, cry22, cry2, cry60, cry64) and two distinct vegetative insecticidal vip4 genes. This combination provides a potential larvicidal and anti-cancer activities to BLB406. It might be a potential solution to some problems such as the narrow insecticidal spectra and insect resistance. The whole BLB406 genome information provides a valuable background for future in silico analyses as well as biotechnological applications in order to increase the production of commercial bioinsecticide based on BLB406 B. thuringiensis strain

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

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Potential of three local marine microalgae from Tunisian coasts for cadmium, lead and chromium removals

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