Purification et Caractérisation de Biomolécules à partir de microorganismes nouvellement isolés et identifiés

Au cours de ce travail de thèse, nous nous sommes intéressés aux études taxonomiques des deux souches TN17 et Fr10 qui sont deux nouvelles espèces du genre Streptomyces dont nous avons proposé les nomenclatures suivantes : Streptomyces lilaceus sp. TN17 et Streptomyces microflavus sp. Fr10. A partir de la souche Streptomyces lilaceus sp. TN17, trois molécules on été purifiées et identifiées par le biais de plusieurs techniques spectroscopiques, il s’agit d’un dérivé de DKP (L-Leu, L-Arg), un dérivé de phtalate le di-(2-éthylhexyl) phtalate et un tértrapeptide cyclique : le 1 - [2 -(cyclopentanecarbonyl-3-phenylpropionyl] – pyrrolidine-2-carboxylique (1-carbamoyl-propyl)-amide. Ces trois molécules présentent des activités antibactériennes et antifongiques. Suite au criblage des souches de bactéries lactiques productrices de bactériocines de la collection de notre laboratoire et leurs caractérisations, nous avons identifié une nouvelle souche de Lactobacillus nommée Lactobacillus plantarum sp.TN635 qui produit une bactériocine « BacTN635 » de 3,8 KDa. Cette dernière a été purifiée à homogénéité, elle possède un spectre d’action très large contre les bactéries à Gram+, à Gram- et contre les champignons filamenteux et unicellulaires. BacTN635 a un effet bactéricide contre Listeria ivanovii BUG 496 et fongistatique contre Candida tropicalis R2 CIP203

Purification et Caractérisation de Biomolécules à partir de microorganismes nouvellement isolés et identifiés

Au cours de ce travail de thèse, nous nous sommes intéressés aux études taxonomiques des deux souches TN17 et Fr10 qui sont deux nouvelles espèces du genre Streptomyces dont nous avons proposé les nomenclatures suivantes : Streptomyces lilaceus sp. TN17 et Streptomyces microflavus sp. Fr10. A partir de la souche Streptomyces lilaceus sp. TN17, trois molécules on été purifiées et identifiées par le biais de plusieurs techniques spectroscopiques, il s’agit d’un dérivé de DKP (L-Leu, L-Arg), un dérivé de phtalate le di-(2-éthylhexyl) phtalate et un tértrapeptide cyclique : le 1 - [2 -(cyclopentanecarbonyl-3-phenylpropionyl] – pyrrolidine-2-carboxylique (1-carbamoyl-propyl)-amide. Ces trois molécules présentent des activités antibactériennes et antifongiques. Suite au criblage des souches de bactéries lactiques productrices de bactériocines de la collection de notre laboratoire et leurs caractérisations, nous avons identifié une nouvelle souche de Lactobacillus nommée Lactobacillus plantarum sp.TN635 qui produit une bactériocine « BacTN635 » de 3,8 KDa. Cette dernière a été purifiée à homogénéité, elle possède un spectre d’action très large contre les bactéries à Gram+, à Gram- et contre les champignons filamenteux et unicellulaires. BacTN635 a un effet bactéricide contre Listeria ivanovii BUG 496 et fongistatique contre Candida tropicalis R2 CIP203. ABSTRACT : In This Thesis, we are interested in taxonomic studies of two strains TN17 and Fr10 which are two new species of the genus Streptomyces, and we have proposed the following names: Streptomyces lilaceus sp. TN17 and Streptomyces microflavus sp. Fr10. From Streptomyces lilaceus strain sp. TN17, three molecules have been purified and identified by means of several spectroscopic techniques, it is a derivative of DKP (L-Leu, Larg), a derivative of phthalate di-(2-ethylhexyl) phthalate and cyclic peptide 1 - [2 - (cyclopentanecarbonyl-3-phenylpropionyl] - pyrrolidine-2-carboxylic acid (1-carbamoylpropyl)- amide. All three molecules exhibit antibacterial and antifungal activities. A novel strain of lactic acid bacteria was isolated and characterized from a collection of our laboratory. It’s identified as a new strain of Lactobacillus, named Lactobacillus plantarum sp.TN635 producing a new bacteriocin "BacTN635" of 3.8 kDa, purified to homogeneity and spectrum with a very broad action against Gram + and Gram-, filamentous and unicellular fungi. BacTN635 has a bactericidal effect against Listeria ivanovii BUG 496 and fungistatic against Candida tropicalis R2 CIP203

Taxonomy and antimicrobial activities of a new Streptomyces sp. TN17 isolated in the soil from an oasis in Tunis

An actinomycete strain referred to as TN17 was screened for its antimicrobial activities. The taxonomic status of this strain was established. The organism was found to have morphological and chemotaxonomic characteristics typical of Streptomycetes. Based on the 16S rRNA nucleotide sequences, Streptomyces sp. TN17 was found to have a relationship with Streptomyces lilaceus, Streptomyces gobitricini and Streptomyces lavendofoliae. Combined analysis of the 16 S rRNA gene sequence (FN687757), phylogenetic analysis, fatty acids profile and physiological tests indicated that there are genotypic and phenotypic differences between TN17 and neighboring Streptomyces species’ neighbors. Therefore, TN17 is a novel species: Streptomyces sp. TN17 (=DSM 42020T=CTM50229T). A cultured extract of this strain inhibits the growth of several Gram positive and Gram negative bacteria and fungi

Purification and structure elucidation of three naturally bioactive molecules from the new terrestrial Streptomyces sp. TN17 strain

Thirty litres of fermentation broth was extracted from the newly isolated Streptomyces sp. strain TN17 and various separation and purification steps led to the isolation of three pure bioactive compounds (1–3). Compound 1: cyclo (L-Leu-L-Arg), a diketopiperazine ‘DKP’ derivative; 2: di-(2-ethylhexyl) phthalate, a phthalate derivative; and 3: cyclo 1-[2-(cyclopentanecarbonyl-3-phenyl-propionyl]-pyrrolidine-2-carboxylic acid (1-carbamoyl-propyl)-amide, a cyclic tetrapeptide derivative. The chemical structure of these three active compounds was established on the basis of spectroscopic studies (MS and NMR) and by comparison with data from the literature. According to our biological studies, the pure compounds (1–3) possess antibacterial and antifungal activities

Zinc oxide nanoparticles in meat packaging: A systematic review of recent literature

Questions have been raised apropos food spoilage, which may pose a great hazard to the global environment and human health. Amongst bio-safe material that retains photocatalysis and impacts, photo-oxidizing on biological and species chemical are ZnO nanoparticles (ZnO-NPs). In this respect, this current paper covers a wide range of topics, starting from the ZnO-NPs synthesis and antibacterial potential to their functional application in meat packaging. A deep comparison of the physicochemical properties of ZnO-NPs synthesized through different routers was reported. In addition, a special focus has been given to antibacterial mechanisms that underlie synthesis parameters. This review also examined the ZnO-NPs impact on the chemico–physico–mechanical properties of the functional coatings/films features. Likewise, the employment of ZnO-NPs in meat packaging was also evaluated. As safer nanoparticles, ZnO-NPs enhance stored meat product quality by microflora growth limitation and retards lipid/protein oxidation. Remarkably, active packaging comprising ZnO-NPs, synthesized or not by plants, showed an eco-friendly solution and future alternative in the meat industry. Information about these topics could help students and scientific researchers who are engaged in chemical engineering, chemistry and meat technology communities to approach the complex thematic of ZnO-NPs.CYTED | Ref.119RT0568GAIN (Axencia Galega de Innovación) | Ref. IN607A2019/01Universidade de Vigo/CISUG

Fluorescence Turns on-off-on Sensing of Ferric Ion and L-Ascorbic Acid by Carbon Quantum Dots

This study used a hydrothermal approach to create a sensitive and focused nanoprobe. Using an “on-off-on” sensing mechanism, the nanoprobe was employed to detect and quantify ferric ions and L-ascorbic acid. Synthesis of the carbon quantum dots was achieved with a single hydrothermal step at 180°C for 24 hours using hot pepper as the starting material. The prepared CQDs showed high fluorescence with a quantum yield of 30% when excited at 350 nm, exhibiting excitation-dependent fluorescence. The emission of the CQDs can be quenched by adding ferric ions, which can be attributed to complex formation leading to nonradiative photoinduced electron transfer (PET). Adding L-ascorbic acid, which can convert ferric ions into ferrous ions, break the complex, and restore the fluorescence of CQD. The linear range and LOD were (10–90) μM and 1 μM for ferric ions, respectively, and L-ascorbic acid’s linear range was (5–100) μM while LOD was 0.1 μM quantification of both substances was accomplished. In addition, orange fruit was used as an actual sample source for ascorbic acid analysis, yielding up to 99% recovery

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

Purification and structure elucidation of biomolecules from novel microorganisms

Au cours de ce travail de thèse, nous nous sommes intéressés aux études taxonomiques des deux souches TN17 et Fr10 qui sont deux nouvelles espèces du genre Streptomyces dont nous avons proposé les nomenclatures suivantes : Streptomyces lilaceus sp. TN17 et Streptomyces microflavus sp. Fr10. A partir de la souche Streptomyces lilaceus sp. TN17, trois molécules on été purifiées et identifiées par le biais de plusieurs techniques spectroscopiques, il s’agit d’un dérivé de DKP (L-Leu, L-Arg), un dérivé de phtalate le di-(2-éthylhexyl) phtalate et un tértrapeptide cyclique : le 1 - [2 -(cyclopentanecarbonyl-3-phenylpropionyl] – pyrrolidine-2-carboxylique (1-carbamoyl-propyl)-amide. Ces trois molécules présentent des activités antibactériennes et antifongiques. Suite au criblage des souches de bactéries lactiques productrices de bactériocines de la collection de notre laboratoire et leurs caractérisations, nous avons identifié une nouvelle souche de Lactobacillus nommée Lactobacillus plantarum sp.TN635 qui produit une bactériocine « BacTN635 » de 3,8 KDa. Cette dernière a été purifiée à homogénéité, elle possède un spectre d’action très large contre les bactéries à Gram+, à Gram- et contre les champignons filamenteux et unicellulaires. BacTN635 a un effet bactéricide contre Listeria ivanovii BUG 496 et fongistatique contre Candida tropicalis R2 CIP203.In This Thesis, we are interested in taxonomic studies of two strains TN17 and Fr10 which are two new species of the genus Streptomyces, and we have proposed the following names: Streptomyces lilaceus sp. TN17 and Streptomyces microflavus sp. Fr10. From Streptomyces lilaceus strain sp. TN17, three molecules have been purified and identified by means of several spectroscopic techniques, it is a derivative of DKP (L-Leu, Larg), a derivative of phthalate di-(2-ethylhexyl) phthalate and cyclic peptide 1 - [2 - (cyclopentanecarbonyl-3-phenylpropionyl] - pyrrolidine-2-carboxylic acid (1-carbamoylpropyl)- amide. All three molecules exhibit antibacterial and antifungal activities. A novel strain of lactic acid bacteria was isolated and characterized from a collection of our laboratory. It’s identified as a new strain of Lactobacillus, named Lactobacillus plantarum sp.TN635 producing a new bacteriocin "BacTN635" of 3.8 kDa, purified to homogeneity and spectrum with a very broad action against Gram + and Gram-, filamentous and unicellular fungi. BacTN635 has a bactericidal effect against Listeria ivanovii BUG 496 and fungistatic against Candida tropicalis R2 CIP203

Chemistry, Safety, and Challenges of the Use of Organic Acids and Their Derivative Salts in Meat Preservation

Meat industries are constantly facing new waves of changes in the consumer’s nutritional trends, food safety, and quality requirements and legislations leading to an increase in interest for meat biopreservation to respond to all of these modern socioeconomic demands. Hence, to replace synthetic and/or expensive additives, new technologies in preserving meat products from microbial contamination have been established. In this context, organic acids and their salts have been considered as the most popular examples of preservatives that offer several advantages to be applied in meat industry. Here, characteristics of organic acids/salts commonly used in meat preservation were described based on the published literature. Moreover, after outlining the challenges and advantages of their use in meat industry, their current applications as meat preservatives on various meat type matrices such as beef, pork, sheep, and poultry were quite exposed based on previous and recent research works. Then, different application types were highlighted. Besides, some potent synergistic approaches based on several combinations of organic acids/salts with different existing preservative techniques are reported with an emphasised discussion of their application as possible solution tools to mainly overcome some problems linked to organic acids/salts when used solely, thus contributing to ensure the overall safety and improve the quality of meats. Finally, despite their usefulness in meat preservation, organic acids/salts may possess detrimental traits. In this context, a detailed discussion on their limits of use in meat products was provided in the last section of this paper