Lobna Elleuch

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.

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.

Purification and characterization of two polyhydroxyalcanoates from Bacillus cereus.

This work aimed to study the potential of 155 strains of Bacillus sp., isolated from a collection of Tunisian microorganisms, for polyhydroxyalcanoates production. The strains were submitted to a battery of standard tests commonly used for determining bioplastic properties. The findings revealed that two of the isolates, namely Bacillus US 163 and US 177, provided red excitations at a wavelength of approximately 543 nm. The polyhydroxyalcanoates produced by the two strains were purified. Gas chromatography-mass spectroscopy (GC-MS), Fourier transformed infrared spectroscopy (FTIR), and gel permeation chromatography (GPC) were used to characterize the two biopolymers. Bacillus US 163 was noted to produce a poly methyl-3-hydroxy tetradecanoic acid (P-3HTD) with an average molecular weight of 455 kDa, a completely amorphous homopolymer without crystallinity. The US 177 strain produced a homopolymer of methyl-3-hydroxy octadecanoic acid (P3-HOD) with an average molecular weight of 555 kDa. Exhibiting the highest performance, US 163 and US 177 were submitted to 16S rRNA gene sequencing, and the results revealed that they belonged to the Bacillus cereus species. Overall, the findings indicated that the Bacilli from petroleum soil have a number of promising properties that make them promising candidates for bioplastic production.

Characterization of the bacteriocin BacJ1 and its effectiveness for the inactivation of Salmonella typhimurium during turkey escalope storage.

During a screening programme for bacteriocin producers, a new lactic acid bacterium called J1, isolated from chicken gizzard, was noted to produce a bacteriocin (BacJ1) that inhibited Gram-positive and Gram-negative food-borne pathogens, especially Salmonella typhimurium, and it was, therefore, selected for identification and further study. The results, from 16S rRNA gene nucleotide sequencing (1583 pb accession no HE861352) and phylogenetic analysis, suggested that the new isolate be assigned as Weissella paramesenteroides J1. BacJ1 was purified from the culture supernatant of the J1 strain using heat treatment (15 min at 90 °C), 80% ammonium sulfate precipitation, and gel filtration (Sephadex G25). MALDI-TOF analyses revealed that BacJ1 had an exact molecular mass of 1881.036 Da. The analysis of the N-terminal sequence (GPAGPFGKLY) of this active peptide revealed no significant similarity to currently available antimicrobial peptides. The addition of pre-purified BacJ1, at a final concentration of 400 AU per gramme, may open new promising opportunities for the prevention of contamination from and growth of pathogenic bacteria, particularly S. typhimurium, during turkey escalope storage at 4 °C.

Efficient role of BacTN635 on the safety properties, sensory attributes, and texture profile of raw minced meat beef and chicken breast.

Bacteriocin BacTN635, produced by Lactobacillus plantarum sp. TN635, was purified and characterised in previous work. In this study we report the biotechnological application of this bacteriocin as a biopreservative during storage at 4°C of raw minced meat beef and chicken breast. Overall, the results obtained showed that the addition of the semi-purified BacTN635 at 500 or 1000 AU g−1 in raw minced meat beef and chicken breast can delay the proliferation of spoilage microorganisms, suppress the growth of the pathogenic microorganism Listeria monocytogenes, improve sensory quality, texture attributes, and extend the shelf-life of these two meat products during refrigerated storage. BacTN635 at 1000 AU g−1 could extend the shelf-life, and the meat showed good sensory characteristics. Therefore, treatment with semi-purified BacTN635 can be used as a safe method for preservation of raw minced meat beef and chicken breast.

The use of newly isolated Streptomyces strain TN258 as potential biocontrol agent of potato tubers leak caused by Pythium ultimum

An actinomycete strain designated TN258, was isolated from Tunisian Sahara soil and selected for its antagonistic activity, especially against Pythium ultimum (P. ultimum) causing potato tubers leak. Based on the results of cultural characteristic of TN258 strain, the 16S rRNA gene nucleotide sequence (1433 bp, accession n° HE600071) and the phylogenetic analysis, we propose the assignment of our new isolate bacterium as Streptomyces TN258 strain. After culture optimization, the inhibitory effect of TN258 free cell supernatant against P. ultimum was evaluated. As result, by application of 50% (v/v) from 25 mg ml−1 of concentration, mycelial growth was totally inhibited with hyphal destruction. At the same concentration, the oospores were distorted and the germination was completely stopped. In potato tubers, Streptomyces TN258 filtrated supernatant, applied 24 h before inoculation by P. ultimum (preventive treatment group) was able to significantly decrease pathogen penetration by 62% and to reduce the percentage of weight loss by 59.43%, in comparison with non‐treated group.