Biology subjects

Dilbaryan, D. S.

Publications and source records attributed to Dilbaryan, D. S..

3 recordsLinked to original sources

bioRxiv2025

Responses of Soil Bacterial Community and Its Resistome to Short-term Exposure to Macrolide Antibiotic Macrolactin A: Metagenomic analysis

An important aspect of studying potential antibacterial biopreparations for crop protection is determining their potential negative impacts on the environment. Plant-associated Bacillus velezensis produce macrolactin A (McA), which determine effectiveness of this bacterial species against numerous human and plant pathogens. However, the effects of McA on the soil microbiome and the selection of specific antibiotic resistance genes (ARGs) among soil bacteria remain unknown. In this study, we use high-throughput sequencing-based metagenomic methods to investigate the differences in structure of the soil bacterial community and the abundance and diversity of ARGs in both McA-treated and untreated samples. The presence of high (10 mg per kg soil) and low (1 mg per kg soil) concentrations of McA induced changes in soil bacterial populations as shown by taxonomic analysis. The relative abundance of Alphaproteobacteria and Betaproteobacteria significantly increased under the McA treatments, while the relative abundance of Thermoleophilia, Rubrobacteria, Planctomycetia and Acidimicrobiia decreased. The ARG profiling results showed that both low and high doses of McA affected ARGs representation in the community. At the same time, a low dose of McA altered the representation of a larger number of ARGs (7 genes) compared to a high dose (3 genes). Overall, exposure to McA consistently altered the abundance of genes associated with resistance to elfamycin, glycopeptide, fluoroquinolone, rifampicin, and macrolide. Correlation analysis identified 185 relationships between 52 antibiotic resistance genes (ARGs) and 34 bacterial genera. Among these bacteria, Streptomyces, Baekduia, and Capillimicrobium were predicted to carry the most diverse ARGs. By assembling and annotating bacterial genomes, we identified the true hosts of ARGs. Chloroflexota were the most prevalent phylum harboring ARGs. Furthermore, profiling the soil microbiomes metabolic potential under low-dose McA revealed increased abundance of genes associated with signaling, chemotaxis, and broad-substrate drug efflux. The collectively obtained data significantly expands the understanding of the functional role of McA in the ecology of Bacillus velezensis, and at the same time provides an assessment of environmental risks associated with the use of biopreparations containing metabolites or living cells of this species bacteria.

microbiology↗

bioRxiv2024

Macrolactin A Is an Inhibitor of Protein Biosynthesis in Bacteria

The macrolide antibiotic, macrolactin A (McA), has been known for its antimicrobial properties since the late 1980s, but the mechanism of its antibacterial activity is still unknown. In this study, we investigated the microbiological and molecular characteristics of McA antimicrobial activity. McA effect on bacteria was found to be both bacteriostatic and bactericidal, depending on species and strains. Regarding the mechanism of action of McA, the following important results were obtained: 1) using in vivo and in vitro systems, we showed that McA is an inhibitor of protein synthesis in bacteria; 2) the concentration of McA required to inhibit protein synthesis in the E. coli cell-free model was found to be 50 times lower than the concentration required in the S. aureus cell-free model; 3) the toe-printing assay revealed that McA inhibits the first step of elongation stage of protein synthesis; 4) we identified single and multiple nucleotide polymorphisms in the gene encoding the translation elongation factor Tu (EF-Tu) by annotating the genomes of McA-resistant Bacillus pumilus McAR and its parental strain. Molecular modeling showed that the McA molecule can form non-covalent bonds with amino acids at the interface of domains 1 and 2 of EF-Tu, characterized by a relatively high docking score. Overall, our study demonstrated that McA acts as an elfamycin-like antibiotic (targeting EF-Tu), addressing a substantial gap in our understanding of the mechanism of action of macrolactin A, a representative member of macrolides.

microbiology↗

bioRxiv2022

Generation of Gausemycin A-resistant Staphylococcus aureus

Gausemycins A and B are the first members of the novel lipoglycopeptides family produced by Streptomyces roseoflavus INA-Ac-5812, which showed the ability to fight clinically important Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus. However, new antibiotics need to be studied in depth to determine their full potential. In this study, we concentrated our efforts to investigate resistance emerging within S. aureus upon gausemycin A application. Using serial passaging of S. aureus FDA209P in increasing concentrations of gausemycin A, we obtained the resistant variant S. aureus 5812R which are 80-times more resistant comparing to the origin strain. Moreover, obtained resistance is stable, since 15 passages in a drug-free medium did not restore bacterial susceptibility to gausemycin A. Elucidating of the differences between resistant and parent strains was concerned antibiotic cross-resistance, structure of bacterial membrane, and response at genetic level. Susceptibility testing of S. aureus 5812R revealed the acquisition of cross-resistance to daptomycin, cefazolin, and tetracycline, while resistance to vancomycin, nisin and ramoplanin absence. The composition of fatty acids constituting the cytoplasmic membrane of S. aureus 5812R, was represented by increased content of anteiso- branched chain fatty acids, while iso-branched chain fatty acids was decreased comparing the origin S. aureus FDA209P strain. The relative expression of the cls gene catalyzing the synthesis of cardiolipin in the resistant cells was higher compared to the S. aureus FDA209P.

microbiology↗