Marine sponges represent a rich source of uncharacterized microbial diversity, and many are host to microorganisms that produce biologically active specialized metabolites. Here, a polyphasic approach was used to characterize two Actinobacteria strains, P01-B04T and P01-F02, that were isolated from the marine sponges Geodia barretti (Bowerbank, 1858) and Antho dichotoma (Esper, 1794), respectively. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains P01-B04T and P01-F02 are closely related to Streptomyces beijiangensis DSM 41794T, Streptomyces laculatispora NRRL B-24909T, and Streptomyces brevispora NRRL B-24910T. The two strains showed nearly identical 16S rRNA gene sequences (99.93%), and the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) relatedness values were 99.96% and 99.6%, respectively, suggesting that these strains are affiliated with the same species. Chemotaxonomic and culture characteristics of both strains were also consistent with the genus Streptomyces, while phenotypic properties, genome-based comparisons, and phylogenomic analyses distinguished strains P01-B04T and P01-F02 from their closest phylogenetic relatives. In silico analysis predicted that the 8.9 Mb genome of P01-B04T contains at least 41 biosynthetic gene clusters (BGCs) encoding secondary metabolites, indicating that this strain could express diverse bioactive metabolites; in support of this prediction, this strain expressed antibacterial activity against Gram-positive bacteria including a clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA) EAMC30. Based on these results, the marine sponge-associated isolates represent a novel species of the genus Streptomyces, for which the name Streptomyces poriferorum sp. nov. is proposed, with P01-B04T (=DSM 111306T = CCM 9048T) as the type strain.
- MeSH
- antibióza * MeSH
- DNA bakterií genetika MeSH
- fylogeneze MeSH
- hybridizace nukleových kyselin MeSH
- methicilin rezistentní Staphylococcus aureus MeSH
- multigenová rodina MeSH
- Porifera * mikrobiologie MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Streptomyces * klasifikace izolace a purifikace MeSH
- techniky typizace bakterií MeSH
- zastoupení bazí MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Streptomyces sp. TR1341 was isolated from the sputum of a man with a history of lung and kidney tuberculosis, recurrent respiratory infections, and COPD. It produces secondary metabolites associated with cytotoxicity and immune response modulation. In this study, we complement our previous results by identifying the genetic features associated with the production of these secondary metabolites and other characteristics that could benefit the strain during its colonization of human tissues (virulence factors, modification of the host immune response, or the production of siderophores). We performed a comparative phylogenetic analysis to identify the genetic features that are shared by environmental isolates and human respiratory pathogens. The results showed a high genomic similarity of Streptomyces sp. TR1341 to the plant-associated Streptomyces sp. endophyte_N2, inferring a soil origin of the strain. Putative virulence genes, such as mammalian cell entry (mce) genes were not detected in the TR1341's genome. The presence of a type VII secretion system, distinct from the ones found in Mycobacterium species, suggests a different colonization strategy than the one used by other actinomycete lung pathogens. We identified a higher diversity of genes related to iron acquisition and demonstrated that the strain produces ferrioxamine B in vitro. These results indicate that TR1341 may have an advantage in colonizing environments that are low in iron, such as human tissue.
- MeSH
- fylogeneze MeSH
- genetické techniky MeSH
- genom genetika MeSH
- geny genetika MeSH
- lidé MeSH
- plíce mikrobiologie MeSH
- počet mikrobiálních kolonií metody MeSH
- Streptomyces * genetika izolace a purifikace MeSH
- tkáně mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Industrial synthetic dyes cause health and environmental problems. This work describes the isolation of 84 bacterial strains from the midgut of the Lasius niger ant and the evaluation of their potential application in dye bioremediation. Strains were identified and classified as judged by rRNA 16S. The most abundant isolates were found to belong to Actinobacteria (49%) and Firmicutes (47.2%). We analyzed the content in laccase, azoreductase and peroxidase activities and their ability to degrade three known dyes (azo, thiazine and anthraquinone) with different chemical structures. Strain Ln26 (identified as Brevibacterium permense) strongly decolorized the three dyes tested at different conditions. Strain Ln78 (Streptomyces ambofaciens) exhibited a high level of activity in the presence of Toluidine Blue (TB). It was determined that 8.5 was the optimal pH for these two strains, the optimal temperature conditions ranged between 22 and 37 °C, and acidic pHs and temperatures around 50 °C caused enzyme inactivation. Finally, the genome of the most promising candidate (Ln26, approximately 4.2 Mb in size) was sequenced. Genes coding for two DyP-type peroxidases, one laccase and one azoreductase were identified and account for the ability of this strain to effectively oxidize a variety of dyes with different chemical structures.
- MeSH
- Actinobacteria enzymologie izolace a purifikace metabolismus MeSH
- Bacteria enzymologie izolace a purifikace metabolismus MeSH
- barvicí látky izolace a purifikace metabolismus MeSH
- biodegradace MeSH
- biotechnologie MeSH
- Brevibacterium enzymologie izolace a purifikace metabolismus MeSH
- Firmicutes enzymologie izolace a purifikace metabolismus MeSH
- Formicidae mikrobiologie MeSH
- lakasa izolace a purifikace metabolismus MeSH
- látky znečišťující životní prostředí izolace a purifikace metabolismus MeSH
- NADH, NADPH oxidoreduktasy izolace a purifikace metabolismus MeSH
- peroxidasa izolace a purifikace metabolismus MeSH
- Streptomyces enzymologie izolace a purifikace metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The main objective of this study was using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for assembling of DSM (German Collection of Microorganisms) Streptomyces spectral database and identification of wild Streptomyces cultures, which were clustered by MALDI-TOF Biotyper OC software as well as for teracycline detection by observing of obtained spectra using flexAnalysis software. Production of tetracycline was confirmed by thin-layer chromatography. Presence of tetracycline mass spectrum was verified by several tetracycline producers (Streptomyces aureofaciens LMG 5968, S. aureofaciens 84/25, and S. aureofaciens BMK) and by pure tetracycline mass. Our results showed that it is possible to use MALDI-TOF MS for identification of tetracycline producers within Streptomyces genera by several easy steps. The purpose of this study was to establish cheap and quick detection of tetracycline producers.
- MeSH
- databáze faktografické MeSH
- lidé MeSH
- rychlé screeningové testy metody MeSH
- software MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice * MeSH
- Streptomyces izolace a purifikace metabolismus MeSH
- tetracyklin chemie izolace a purifikace metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Mutualistic associations between insects and microorganisms must imply gains for both partners, and the emphasis has mostly focused on coevolved host-symbiont systems. However, some insect hosts may have evolved traits that allow for various means of association with opportunistic microbial communities, especially when the microbes are omnipresent in their environment. It was previously shown that colonies of the subterranean termite Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) build nests out of fecal material that host a community of Streptomyces Waksman and Henrici (Actinomycetales: Streptomycetaceae). These Actinobacteria produce an array of bioactive metabolites that provides a level of protection for termites against certain entomopathogenic fungi. How C. formosanus acquires and maintains this association remains unknown. This study shows that the majority of Streptomyces isolates found in field termite fecal nest materials are identical to Streptomyces isolates from soils surrounding the nests and are not vertically inherited. A survey of Streptomyces communities from C. formosanus fecal nest materials sampled at 20 locations around the world revealed that all nests are reliably associated with a diverse Streptomyces community. The C. formosanus fecal nest material therefore provides a nutritional framework that can recruit beneficial Streptomyces from the soil environment, in the absence of long-term coevolutionary processes. A diverse Streptomyces community is reliably present in soils, and subterranean termite colonies can acquire such facultative symbionts each social cycle into their fecal nest. This association probably emerged as an exaptation from the existing termite nest structure and benefits both the termite and the opportunistic colonizing bacteria.
- MeSH
- antifungální látky analýza MeSH
- feces mikrobiologie MeSH
- Isoptera mikrobiologie MeSH
- půdní mikrobiologie * MeSH
- Streptomyces chemie izolace a purifikace MeSH
- symbióza MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Isolation of unexplored frontier molecules are needed to treat multidrug resistant pathogens especially Methicillin resistant Staphylococcus aureus (MRSA). A marine sponge endosymbiotic Streptomyces albus ICN33 produces an anti-MRSA metabolite is reported. The crude extract exhibited anti-MRSA activity and the active principle was isolated through fermentation and chromatographic techniques. A compound PVI331 with a molecular mass of 506 Da have been determined by high resolution mass spectrometry. LC–MS based dereplication analysis had revealed that the detected compound PVI331 as unknown. The antibacterial assay of the compound PVI331 showed remarkable antagonistic activity against MRSA and Escherichia coli. Minimum inhibitory concentrations were found to be 1 μg/ml against MRSA. Sub-inhibitory concentration of the compound PVI331 reduced the biofilm formation of Staphylococcus aureus ATCC25923 and increased the cell surface hydrophobicity index. Scanning electron microscopic observation of the sub-inhibitory concentration exposure revealed a wrinkled membrane surface and slight cellular damage shows the cell wall distracting property of the compound. Zebrafish embryo based toxicity assays exhibited 48 ± 2 μg/ml of LC50 value and 30 μg/ml of compound as maximal non-lethal concentration which had demonstrated the positive relationship in safety index. This study highlighted the anti-MRSA property of Streptomyces albus ICN33 from a marine sponge.
- MeSH
- antibakteriální látky izolace a purifikace terapeutické užití MeSH
- biofilmy účinky léků MeSH
- dánio pruhované MeSH
- embryo nesavčí účinky léků MeSH
- methicilin rezistentní Staphylococcus aureus * metabolismus patogenita účinky léků MeSH
- Porifera mikrobiologie MeSH
- Streptomyces chemie izolace a purifikace metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Publikační typ
- práce podpořená grantem MeSH
While it is known that several Actinobacteria produce enzymes that decompose polysaccharides or phenolic compounds in dead plant biomass, the occurrence of these traits in the environment remains largely unclear. The aim of this work was to screen isolated actinobacterial strains to explore their ability to produce extracellular enzymes that participate in the degradation of polysaccharides and their ability to cometabolically transform phenolic compounds of various complexities. Actinobacterial strains were isolated from meadow and forest soils and screened for their ability to grow on lignocellulose. The potential to transform (14)C-labelled phenolic substrates (dehydrogenation polymer (DHP), lignin and catechol) and to produce a range of extracellular, hydrolytic enzymes was investigated in three strains of Streptomyces spp. that possessed high lignocellulose degrading activity. Isolated strains showed high variation in their ability to produce cellulose- and hemicellulose-degrading enzymes and were able to mineralise up to 1.1% and to solubilise up to 4% of poplar lignin and to mineralise up to 11.4% and to solubilise up to 64% of catechol, while only minimal mineralisation of DHP was observed. The results confirm the potential importance of Actinobacteria in lignocellulose degradation, although it is likely that the decomposition of biopolymers is limited to strains that represent only a minor portion of the entire community, while the range of simple, carbon-containing compounds that serve as sources for actinobacterial growth is relatively wide.
- MeSH
- bakteriální proteiny biosyntéza MeSH
- beta-glukosidasa biosyntéza MeSH
- biodegradace MeSH
- biomasa MeSH
- celulosa-1,4-beta-cellobiosidasa biosyntéza MeSH
- celulosa metabolismus MeSH
- hydrolýza MeSH
- katecholy metabolismus MeSH
- kinetika MeSH
- lignin metabolismus MeSH
- Populus chemie MeSH
- půdní mikrobiologie * MeSH
- radioizotopy uhlíku MeSH
- Streptomyces enzymologie izolace a purifikace MeSH
- stromy chemie MeSH
- xylosidasy biosyntéza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The genetic diversity of streptomycetes in colliery spoil heaps (Sokolov, Czech Republic) was investigated by restriction pattern analysis of 16S-internal transcribed spacer rDNA and 16S sequences. We sampled freshly excavated Miocene sediment (17-19-million-year-old) and four sites of primary succession (initial, early, middle, and late stages; aged 1-44 years) on the same sediment. Active bacteria were present even in fresh Miocene sediment, and the relative proportion of actinomycetes among total bacterial and their genetic diversity increased significantly with the age of the sampling site. The replacement of pioneer species by late succession species during succession was observed. Plate assays of Streptomyces strains revealed 27% antibiotic-producing strains. Screening for nonribosomal peptide synthases and type I polyketide synthases systems suggested that 90% and 55% streptomycetes, respectively, are putative producers of biologically active compounds. The frequencies of tetracycline-, amoxicillin-, and chloramphenicol-resistant streptomycetes were 6%, 9%, and 15%, respectively. These findings document the occurrence of genetic elements encoding antibiotic resistance genes and the production of antibiotics by streptomycetes located in pristine environments. Our results indicate key roles for ancient streptomycetes related to S. microflavus, S. spororaveus, and S. flavofuscus in pioneering community development in freshly excavated substrates.
- MeSH
- antibakteriální látky metabolismus MeSH
- biodiverzita MeSH
- DNA bakterií genetika MeSH
- fylogeneze MeSH
- geologické sedimenty mikrobiologie MeSH
- hornictví MeSH
- molekulární sekvence - údaje MeSH
- RNA ribozomální 16S genetika MeSH
- Streptomyces klasifikace genetika izolace a purifikace metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Aim: Identifi cation of cyclopropyl fatty acids in walnut oil.Method: GC/MS method was developed for the determination of eight cyclopropyl fatty acids in walnut (Juglansregia) oil. Results: Monocyclopropane acids: methyl 9-cyclopropyl-nonanoate, 6,7-methylene-, 8,9-methylene-, 9,10-methylene-,11,12-methylene octadecanoates, and dicyclic acid – methyl 9,10,12,13-dimethylene octadecanoate, tricyclic acid– methyl 9,10,12,13,15,16-trimethylene octadecanoate, and unsaturated - methyl 2-octylcyclopropene-1-octanoate weredetected in walnut oil by GC-MS and their mass spectra studied. Four cyclic fatty acids were identifi ed for the fi sttime in seed oils. Conclusions: Eight cyclopropyl fatty acids were detected in the Mediterranean nuts for the fi rst time.
- MeSH
- Bacteria enzymologie izolace a purifikace MeSH
- cyklopropany izolace a purifikace MeSH
- estery MeSH
- hmotnostní spektrometrie metody využití MeSH
- Juglans MeSH
- kyseliny linolenové izolace a purifikace MeSH
- nenasycené mastné kyseliny izolace a purifikace MeSH
- oleje rostlin MeSH
- rostliny MeSH
- sluneční záření MeSH
- Streptomyces enzymologie izolace a purifikace MeSH
A DNA fragment containing part of the salinomycin biosynthetic gene cluster from industrial strain Streptomyces albus CCM 4719 was cloned. Sequence analysis of the 25.809-kbp fragment revealed the presence of 8 open reading frames (ORFs), including two large ORFs encoding three modular sets of oligoketide synthase, followed by three genes (salRI, salRII, salRIII) encoding transcriptional regulators. The first two regulators, SalRI and SalRII, belonged to the novel LAL family of large transcriptional regulators. SalRIII was highly similar to the NysRIV, AmphRIV, and FscRI transcriptional regulators from the oligoene macrolides nystatin, amphotericin, and R008/candicidin clusters, respectively.
- MeSH
- finanční podpora výzkumu jako téma MeSH
- genomová knihovna MeSH
- klonování DNA metody MeSH
- otevřené čtecí rámce genetika MeSH
- polymerázová řetězová reakce metody využití MeSH
- pyrany izolace a purifikace MeSH
- regulační geny genetika MeSH
- sekvenční analýza DNA metody využití MeSH
- Streptomyces genetika izolace a purifikace růst a vývoj MeSH