Effective biosynthesis of benzoyl-pyrrothine dithiolopyrrolone antibiotic by cinnamic acid-precursor addition in culture of Saccharothrix algeriensis NRRL B-24137.

Dithiolopyrrolone antibiotics, produced by several micro-organisms, are known for their strong antimicrobial and antitumor activities. Among of this micro-organisms, Saccharothrix algeriensis NRRL B-24137, a rare actinobacterium, has the ability to produce several dithiolopyrrolones derivatives depending on precursors added in the culture medium. After 10 days of strain fermentation on semi-synthetic medium supplemented with cinnamic acid and HPLC purification, biosynthesis of benzoyl-pyrrothine dithiolopyrrolone was evidenced through complete spectroscopic (UV-visible and 1H and 13C NMR) and spectrometric (electron impact mass spectrum) analyses. The pure molecule showed appreciable minimum inhibitory concentration values against several Gram-positive bacteria and filamentous fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Dithiolopyrrolone antibiotics, known for their strong antimicrobial activities, gained greater interest after the discovery of their antitumor properties. Depending on precursors added, Saccharothrix algeriensis NRRL B-24137 has the ability to produce several dithiolopyrrolones derivatives. Since biological activities of dithiolopyrrolones are related to their variable structure, discover of new natural analogues to be therapeutically explored remains a significant framework of research. In this study, a new dithiolopyrrolone derivative was purified from the fermentation broth of S. algeriensis NRRL B-24137. This new antibiotic, characterized as benzoyl-pyrrothine dithiolopyrrolone, was induced by adding cinnamic acid, as precursor, to a semi-synthetic medium.

A new dithiolopyrrolone antibiotic triggered by a long fermentation of Saccharothrix algeriensis NRRL B-24137 in sorbic acid-amended medium.

Saccharothrix algeriensis NRRL B-24137 is an actinobacterium isolated from Algerian Saharan soil. It produces bioactive compounds belonging to the dithiolopyrrolone class of antibiotics, which are characterized by the possession of a unique pyrrolinonodithiole nucleus. Dithiolopyrrolones are known for their strong antibacterial and antifungal activities. This class of antibiotics generated great interest after the discovery of their anticancer properties. In this study, an antibiotic named PR11, produced after a long bacterial fermentation (11 days) in sorbic acid-containing culture broth, was characterized as a new dithiolopyrrolone derivative. After HPLC analysis and purification, the chemical structure of this antibiotic was determined by 1 H- and 13 C-nuclear magnetic resonance, mass and UV-visible data. PR11 was thus characterized as an iso-hexanoyl-pyrrothine, a novel dithiolopyrrolone derivative. The minimum inhibitory concentrations of the new induced antibiotic were determined against several pathogenic micro-organisms. A moderate to strong activity was noted against all Gram-positive bacteria, filamentous fungi and yeasts tested. SIGNIFICANCE AND IMPACT OF THE STUDY: Given the strong activities of dithiolopyrrolones against diverse prokaryotic and eukaryotic micro-organisms including potent selective-anticancer activity, the discovery of new-related derivatives draw continuous attention for therapeutic research. Depending on nature and concentration of added precursor, Saccharothrix algeriensis NRRL B-24137 produce several dithiolopyrrolone coumpounds. In this study, sorbic acid addition combined to long fermentation duration was shown to induce the biosynthesis of a novel dithiolopyrrolone derivative. After purification and full spectroscopic and spectrometric study, the compound was characterized as iso-hexanoyl-pyrrothine. In the future investigation for novel dithiolopyrrolone discovery, fermentation duration should be regarded as a key parameter as well.

Insecticidal activity of a Moroccan strain of Streptomyces phaeochromogenes LD-37 on larvae, pupae and adults of the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae).

The Mediterranean fruit fly (medfly), Ceratitis capitata, is considered the most important fruit pest worldwide. Its management is mainly based on the use of chemical insecticides. Although these conventional pesticides are effective at high doses, they cause considerable human health and environment problems. Thus, the aim of this study was to assess insecticidal activity of Moroccan actinobacteria against C. capitata. A total of 12 preselected actinobacteria isolated from various Moroccan habitats were screened for their insecticidal activity against larvae, pupae and adults of C. capitata. Four actinobacteria isolates were significantly active against the first-instar larvae, and nine were active against the medfly adult, while no significant mortality was obtained against the third-instar larval and pupal stages. Among the selected isolates, the biological screening revealed that strain Streptomyces LD-37, which showed 99.4% similarity with Streptomyces phaeochromogenes, exhibited the maximal corrected larval mortality of 98%. Moreover, the isolates AS1 and LD-37 showed the maximum significant corrected mortality against adults of 32.5 and 28.2%, respectively. The crude extract obtained from a fermented culture of strain S. phaeochromogenes LD-37 was separated into six fractions by thin layer chromatography. Fractions F3 and F4 caused a significant corrected larval mortality of 66.7 and 53.3%, respectively; whereas the maximum reduction in adult emergence was obtained with fraction F4. This finding could be useful for utilizing S. phaeochromogenes LD-37 as an alternative to chemical insecticides in pest management of C. capitata.

A novel hydroxamic acid-containing antibiotic produced by a Saharan soil-living Streptomyces strain.

During screening for potentially antimicrobial actinobacteria, a highly antagonistic strain, designated WAB9, was isolated from a Saharan soil of Algeria. A polyphasic approach characterized the strain taxonomically as a member of the genus Streptomyces. The strain WAB9 exhibited a broad spectrum of antimicrobial activity toward various multidrug-resistant micro-organisms. A PCR-based assay of genomic potential for producing bioactive metabolites revealed the presence of PKS-II gene. After 6 days of strain fermentation, one bioactive compound was extracted from the remaining aqueous phase and then purified by HPLC. The chemical structure of the compound was determined by spectroscopic (UV-visible, and (1)H and (13)C NMR) and spectrometric analysis. The compound was identified to be 2-amino-N-(2-amino-3-phenylpropanoyl)-N-hydroxy-3-phenylpropanamide, a novel hydroxamic acid-containing molecule. The pure molecule showed appreciable minimum inhibitory concentration values against a selection of drug-resistant bacteria, filamentous fungi and yeasts. Significance and impact of the study: This study presents the isolation of a Streptomyces strain, named WAB9, from a Saharan soil in Algeria. This strain was found to produce a new hydroxamic acid-containing molecule with interesting antimicrobial activities towards various multidrug-resistant micro-organisms. Although hydroxamic acid-containing molecules are known to exhibit low toxicities in general, only real evaluations of the toxicity levels could decide on the applications for which this new molecule is potentially most appropriate. Thus, this article provides a new framework of research.

Characterization and antagonistic properties of Streptomyces strains isolated from Saharan soils, and evaluation of their ability to control seedling blight of barley caused by Fusarium culmorum.

UNLABELLED: During a screening for potential plant disease control actinomycetes, a total of 133 strains were isolated from Saharan soil samples of seven Algerian regions by dilution technique on chitin-vitamins agar medium. Screening for antagonistic properties using streak assay method showed that 25% of isolates demonstrated strong activities against a wide range of plant pathogenic fungi. Due to their strong anti-Fusarium activities, six of these isolates were selected and subsequently related to Streptomyces species by polyphasic analysis. These isolates were evaluated for their biocontrol ability against Fusarium culmorum, a serious pathogenic fungus of cereals crops related to damping-off and seedling blight resulting in yield loss. Barley seeds were chosen as cereal plant model. Surface bacterized seeds with TW3, RI3 and TW2 strains expressed the highest performances and permit to reduce significantly both the disease occurrence on seedlings (62-76%) and the extent of seedling blight symptoms (over than 95%). However, a negative effect on plant establishment was observed for RI3 treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The genus Fusarium is considered to be one of the most problematic phytopathogenic fungi for crop culture worldwide. Inside this genus, F. culmorum is the aetiological agent of seedling blight in various monocotyledonous plants such as barley and cause extensive yield and quality losses in humid and semi-humid regions. Biological control may be a successful alternative to chemical control, particularly with the controversy surrounding the use of the fungicides and the limited obtained results to control F. culmorum. This study highlights the effectiveness of some antagonistic Streptomyces isolated from Algerian Saharan soils to control F. culmorum by the reduction in disease occurrence and disease severity suggesting their use on microbial biocontrol formulation against soilborne diseases.

Activity of 2,4-Di-tert-butylphenol produced by a strain of Streptomyces mutabilis isolated from a Saharan soil against Candida albicans and other pathogenic fungi.

In a search for new antifungal antibiotics active against Candida albicans and others pathogenic fungi, a strain of actinobacteria, designated G61, was isolated from a Saharan soil and tested for its activity against these microorganisms. The analysis of its 16S rDNA sequence showed a similarity level of 100% with Streptomyces mutabilis NBRC 12800(T). The highest anticandidal activities produced by the strain G61 were obtained on Bennett medium in the fourth day of incubation. The active product, extracted by n-butanol, contained one bioactive spot detected on thin layer chromatography plates. It was purified by HPLC and its chemical structure was determined by spectroscopic analyses as 2,4-Di-tert-butylphenol. The minimum inhibitory concentrations (MIC) of this product against several strains of pathogenic microorganisms are interesting.

Antimicrobial activity of saquayamycins produced by Streptomyces spp. PAL114 isolated from a Saharan soil.

A new strain of actinomycete designated PAL114, producing antimicrobial compounds, was isolated from a Saharan soil in Ghardaïa, Algeria. Morphological and chemical studies showed that this strain belonged to the genus Streptomyces. Two bioactive compounds, named P41A and P41B, were extracted by dichloromethane from the cell-free supernatant broth of strain PAL114 and were purified by HPLC. Minimum inhibitory concentrations of the pure antibiotics were determined against yeasts, filamentous fungi and bacteria, most of which are pathogenic or toxigenic for human and multiresistant to antibiotics. The strongest activities were observed against Candida albicans M3 and Bacillus subtilis ATCC 6633. The chemical structures of the compounds were determined by spectroscopic analysis of UV-visible and 1H and 13C NMR spectra and spectrometric analysis of mass spectrum. The compounds P41A and P41B were identified as saquayamycins A and C, respectively. These compounds belong to the aquayamycin-group antibiotics, which are known in the literature for their anticancer and antibacterial activities.

A 31 T split-pair pulsed magnet for single crystal x-ray diffraction at low temperature.

We have developed a pulsed magnet system with panoramic access for synchrotron x-ray diffraction in magnetic fields up to 31 T and at low temperature down to 1.5 K. The apparatus consists of a split-pair magnet, a liquid nitrogen bath to cool the pulsed coil, and a helium cryostat allowing sample temperatures from 1.5 up to 250 K. Using a 1.15 MJ mobile generator, magnetic field pulses of 60 ms length were generated in the magnet, with a rise time of 16.5 ms and a repetition rate of 2 pulses/h at 31 T. The setup was validated for single crystal diffraction on the ESRF beamline ID06.

Taxonomy and chemical characterization of new antibiotics produced by Saccharothrix SA198 isolated from a Saharan soil.

Actinomycete strain SA198, isolated from a Saharan soil sample of Algeria, exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria, and phytopathogenic and toxinogenic fungi. The morphological and chemotaxonomic characteristics of the strain were consistent with those of the genus Saccharothrix. Analysis of the 16S rRNA gene sequence of strain SA198 showed a similarity level ranging between 97.2 and 98.8% within Saccharothrix species, S. australiensis being the most closely related. Two new active products were isolated by reverse HPLC using a C18 column. The ultraviolet-visible (UV-VIS), infrared (IR), mass, and (1)H and (14)C nuclear magnetic resonance (NMR) spectra showed that these products were new bioactive compounds. The minimum inhibitory concentrations of these antibiotics showed a strong activity against fungi and moderate activities against Gram-positive and Gram-negative bacteria.

[Antimicrobial activity of a Saharan Streptomyces spp. PAL111 strain against various clinical and toxinogenic microorganisms resistant to antibiotics]. / Activité antimicrobienne de Streptomyces sp. PAL111 d'origine saharienne contre divers microorganismes cliniques et toxinogènes résistants aux antibiotiques.

OBJECTIVE: Study of the taxonomy and the biological activity of the actinomycete strain PAL111 against several pathogenic and toxigenic microorganisms for humans, and resistant to many antibiotics. MATERIALS AND METHODS: The taxonomic study of isolate PAL111 is carried out on the basis of phenotypic and molecular characteristics. The tests against the pathogenic microorganisms are realized on ISP-2 and Bennett media. The kinetics of antibiotic production was investigated on ISP-2 medium. The antibiotic is highlighted by bioautography and chemical revelations, and then purified by chromatography on thick layer of silica gel and Sephadex LH20 column. The minimum inhibitory concentrations (MIC) were determined against pathogenic microorganisms. RESULTS: The phenotypic and molecular studies showed that the isolate PAL111 is closely related to the type strain of Streptomyces ambofaciens. It showed a strong activity against Candida albicans, filamentous fungi, and Gram-positive and Gram-negative bacteria. The optimal antibiotic production was observed at the end of the exponential phase of growth and at the beginning of the decline phase. The bioautography tests showed the presence of an antibiotic with both antibacterial and antifungal activities. This antibiotic is a hydrophilic amino-glycoside compound. The MIC were observed between 2 and 20µg/mL for yeasts, 10 and 50µg/mL for filamentous fungi, 2 and 10µg/mL for Gram-positive bacteria, and 20 and 75µg/mL for Gram-negative bacteria. CONCLUSION: The strong activity of isolate PAL111 against the pathogenic microorganisms and the polar characteristic of the produced antibiotic could encourage further studies on this bioactive molecule.

A 30 T pulsed magnet with conical bore for synchrotron powder diffraction.

We report on the design, construction, and operation of a horizontal field, 30 T magnet system with a conical bore optimized for synchrotron x-ray powder diffraction. The magnet offers ±31° optical access downstream of the sample, which allows to measure a sufficiently large number of Debye rings for an accurate crystal structure analysis. Combined with a 290 kJ generator, magnetic field pulses of 60 ms length were generated in the magnet, with a rise time of 4.1 ms and a repetition rate of 6 pulses/h at 30 T. The coil is mounted inside a liquid nitrogen bath. A liquid helium flow cryostat reaches into the coil and allows sample temperature between 5 and 250 K. The setup was used on the European Synchrotron Radiation Facility beamlines ID20 and ID06.

de Haas-van Alphen oscillations in the underdoped high-temperature superconductor YBa2Cu3O6.5.

The de Haas-van Alphen effect was observed in the underdoped cuprate YBa2Cu3O6.5 via a torque technique in pulsed magnetic fields up to 59 T. Above a field of approximately 30 T the magnetization exhibits clear quantum oscillations with a single frequency of 540 T and a cyclotron mass of 1.76 times the free electron mass, in excellent agreement with previously observed Shubnikov-de Haas oscillations. The oscillations obey the standard Lifshitz-Kosevich formula of Fermi-liquid theory. This thermodynamic observation of quantum oscillations confirms the existence of a well-defined, closed, and coherent, Fermi surface in the pseudogap phase of cuprates.

Isolation and partial characterization of pigment-like antibiotics produced by a new strain of Streptosporangium isolated from an Algerian soil.

AIMS: Identification of a new actinomycete strain Sg3, belonging to the genus Streptosporangium and partial characterization of the produced antibacterial activities. METHODS AND RESULTS: The strain Sg3 was isolated from an Algerian Saharan soil and identified by morphological, chemotaxonomic and phylogenetic analyses to the genus Streptosporangium. The comparison of its physiological characteristics with those of known species of Streptosporangium showed significant differences with the nearest species Streptosporangium carneum. Analysis of the 16S rDNA sequence of strain Sg3 showed a similarity level ranging between 97% and 98.8% within Streptosporangium species, with S. carneum the most closely related. Strain Sg3 showed a red coloured antibacterial activity against gram-positive bacteria on several culture media. The purification of the red pigment by chromatographic methods led to the isolation of three active products. The (1)H nuclear magnetic resonance (NMR), mass, infrared (IR) and ultraviolet-visible (UV-VIS) data of these molecules strongly suggested that they belonged to the quinone-anthracycline group with three or more rings. CONCLUSIONS: Strain Sg3 represents a distinct phyletic line suggesting a new genomic species. It produces antibacterial activities identified as quinone-anthracycline aromatics. SIGNIFICANCE AND IMPACT OF THE STUDY: The quinone-anthracycline antibiotics are known for their antimicrobial and antineoplastic activities and are used in chemotherapy for the treatment of many cancer diseases. The present work constitutes the first stage of a whole series of studies to be realized on these antibiotics before arriving at a possible application.

Antimicrobial compounds produced by Actinomadura sp. AC104 isolated from an Algerian Saharan soil.

During a search for nonpolyenic antifungal antibiotics, an actinomycete designated AC104 was isolated from a Saharan soil sample by a dilution agar plating method using a chitin - vitamins B medium supplemented with rifampicin. Isolate AC104 presented the morphological and the chemical characteristics of the genus Actinomadura. On the basis of 76 physiological tests and 16S rDNA analysis, this isolate was determined to be quite different from the known species of Actinomadura. It is active against filamentous fungi and both Gram-positive and Gram-negative bacteria. The production of antibiotic substances was investigated using several culture media. The highest antimicrobial activities were obtained on ISP2 medium. The benzenic extract contained five bioactive spots detected on thin layer chromatography plates. Among these antibiotics, a complex called 104A, which showed the more interesting antifungal activity, was selected and purified by reverse-phase high-pressure liquid chromatography. This complex is composed of four compounds. Ultraviolet-visible, infrared, mass, and 1H nuclear magnetic resonance spectroscopy studies showed that these molecules contain an aromatic ring substituted by aliphatic chains. These compounds differ from the known antibiotics produced by Actinomadura species.

Saccharothrix algeriensis sp. nov., isolated from Saharan soil.

The taxonomic position of a soil isolate, strain SA 233T, recovered from Saharan soil from Algeria was established using a polyphasic approach. This isolate has been previously reported to produce three novel dithiolopyrrolone antibiotics, and preliminary chemotaxonomic and morphological characteristics suggested that it was representative of a member of the genus Saccharothrix. Phylogenetic analysis of the strain from 16S rDNA sequences, along with a detailed analysis of morphological, chemotaxonomic and physiological characteristics, indicates that it belongs to the genus Saccharothrix and represents a novel species that is readily distinguished from all recognized Saccharothrix species. The name Saccharothrix algeriensis sp. nov. is proposed for the isolate, with type strain SA 233T (=NRRL B-24137T=DSM 44581T).