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
Marine sponges, a well-documented prolific source of natural products, harbor highly diverse microbial communities. Their extracts were previously shown to contain quorum sensing (QS) signal molecules of the N-acyl homoserine lactone (AHL) type, known to orchestrate bacterial gene regulation. Some bacteria and eukaryotic organisms are known to produce molecules that can interfere with QS signaling, thus affecting microbial genetic regulation and function. In the present study, we established the production of both QS signal molecules as well as QS inhibitory (QSI) molecules in the sponge species Sarcotragus spinosulus. A total of eighteen saturated acyl chain AHLs were identified along with six unsaturated acyl chain AHLs. Bioassay-guided purification led to the isolation of two brominated metabolites with QSI activity. The structures of these compounds were elucidated by comparative spectral analysis of 1HNMR and HR-MS data and were identified as 3-bromo-4-methoxyphenethylamine (1) and 5,6-dibromo-N,N-dimethyltryptamine (2). The QSI activity of compounds 1 and 2 was evaluated using reporter gene assays for long- and short-chain AHL signals (Escherichia coli pSB1075 and E. coli pSB401, respectively). QSI activity was further confirmed by measuring dose-dependent inhibition of proteolytic activity and pyocyanin production in Pseudomonas aeruginosa PAO1. The obtained results show the coexistence of QS and QSI in S. spinosulus, a complex signal network that may mediate the orchestrated function of the microbiome within the sponge holobiont.
- MeSH
- Escherichia coli účinky léků fyziologie MeSH
- faktory virulence MeSH
- fylogeneze MeSH
- luminiscenční měření MeSH
- Porifera genetika metabolismus mikrobiologie MeSH
- proteasy chemie farmakologie MeSH
- pyokyanin chemie farmakologie MeSH
- quorum sensing účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The high demand for new antibacterials fosters the isolation of new biologically active compounds producing actinobacteria. Here, we report the isolation and initial characterization of cultured actinobacteria from dominant benthic organisms' communities of Lake Baikal. Twenty-five distinct strains were obtained from 5 species of Baikal endemic macroinvertebrates of amphipods, freshwater sponges, turbellaria worms, and insects (caddisfly larvae). The 16S ribosomal RNA (rRNA)-based phylogenic analysis of obtained strains showed their affiliation to Streptomyces, Nocardia, Pseudonocardia, Micromonospora, Aeromicrobium, and Agromyces genera, revealing the diversity of actinobacteria associated with the benthic organisms of Lake Baikal. The biological activity assays showed that 24 out of 25 strains are producing compounds active against at least one of the test cultures used, including Gram-negative bacteria and Candida albicans. Complete dereplication of secondary metabolite profiles of two isolated strains led to identification of only few known compounds, while the majority of detected metabolites are not listed in existing antibiotic databases.
- MeSH
- Actinobacteria chemie klasifikace izolace a purifikace metabolismus MeSH
- Amphipoda mikrobiologie MeSH
- antiinfekční látky chemie metabolismus MeSH
- Bacteria účinky léků MeSH
- bezobratlí mikrobiologie MeSH
- fylogeneze MeSH
- hmyz mikrobiologie MeSH
- houby účinky léků MeSH
- jezera mikrobiologie MeSH
- Porifera mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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
