Characterization of Bacillus velezensis EV17 and K-3618 and Their Polyketide Antibiotic Oxydifficidin, an Inhibitor of Prokaryotic Translation with Low Cytotoxicity

Oxydifficidin is a natural polyketide antibiotic that has long been recognized as a ribosome-targeting agent that inhibits protein synthesis. In this paper, we describe <i>Bacillus velezensis</i> strain EV17 and compare its complete genome sequence with that of the previously characterized <i>B. velezensis</i> strain K-3618 and the difficidin biosynthetic gene cluster (BGC) combined with mass spectrometry to elucidate the production of oxydifficidin by these strains. Toeprinting and small fluorescent peptide assays showed that isolated oxydifficidin induces a generalized inhibition of translation at every step of elongation in protein biosynthesis. In previous studies, it has been demonstrated that oxydifficidin targets bL12 protein. Although spontaneous mutations conferring resistance to oxydifficidin in ribosomal protein bL12 located relatively close to the thiostrepton binding site on uL11, our data show that oxydifficidin binding does not interfere with thiostrepton, thereby refining previous findings about its putative ribosomal target. We are the first to show that this compound does not affect eukaryotic translation and has two orders of magnitude lower effect on eukaryotic cells compared to bacteria. These facts are important to further investigate its potential as a bioprotectant against phytopathogens or even as a therapeutic agent.

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Publication Details

Journal
International Journal of Molecular Sciences
Published
2025-12-05
DOI
https://doi.org/10.3390/ijms262411777
Primary Topic
RNA and protein synthesis mechanisms
Type
article
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article

Characterization of Bacillus velezensis EV17 and K-3618 and Their Polyketide Antibiotic Oxydifficidin, an Inhibitor of Prokaryotic Translation with Low Cytotoxicity

Elena B. Guglya, Mikhail V. Biryukov, Arsen M. Kudzhaev, Stanislav S. Terekhov et al.
International Journal of Molecular Sciences
RNA and protein synthesis mechanisms
article

Characterization of Bacillus velezensis EV17 and K-3618 and Their Polyketide Antibiotic Oxydifficidin, an Inhibitor of Prokaryotic Translation with Low Cytotoxicity

Elena B. Guglya, Mikhail V. Biryukov, Arsen M. Kudzhaev, Stanislav S. Terekhov, Петр В. Сергиев, Sergey I. Kovalchuk, Kirill S. Antonets, Vera A. Alferova, Maria G. Khrenova, Vladimir I. Polshakov, Maria I. Zvereva, Vladimir K. Chebotar, Dmitrii A. Lukianov, Anton A. Nizhnikov, Tatiana V. Panova, Olga A. Belozerova, A. Yu. Rudenko, Anton E. Shikov, Maria E. Baganova, Vadim N. Tashlitsky, Nikolai D. Dagaev, Margarita N. Baranova, Natalia V. Sumbatyan, Maria S. Gancheva, Andrey G. Tereshchenkov, Yuliya V. Zakalyukina, E.G. Yurchenko, Maria N. Romanenko, Vladislava E. Sagitova, O. Dontsova, Alisa P. Chernyshova, Valeriya I. Marina, Maksim A. Kryakvin, Kseniya A. Arzamazova
article en

Abstract

Oxydifficidin is a natural polyketide antibiotic that has long been recognized as a ribosome-targeting agent that inhibits protein synthesis. In this paper, we describe <i>Bacillus velezensis</i> strain EV17 and compare its complete genome sequence with that of the previously characterized <i>B. velezensis</i> strain K-3618 and the difficidin biosynthetic gene cluster (BGC) combined with mass spectrometry to elucidate the production of oxydifficidin by these strains. Toeprinting and small fluorescent peptide assays showed that isolated oxydifficidin induces a generalized inhibition of translation at every step of elongation in protein biosynthesis. In previous studies, it has been demonstrated that oxydifficidin targets bL12 protein. Although spontaneous mutations conferring resistance to oxydifficidin in ribosomal protein bL12 located relatively close to the thiostrepton binding site on uL11, our data show that oxydifficidin binding does not interfere with thiostrepton, thereby refining previous findings about its putative ribosomal target. We are the first to show that this compound does not affect eukaryotic translation and has two orders of magnitude lower effect on eukaryotic cells compared to bacteria. These facts are important to further investigate its potential as a bioprotectant against phytopathogens or even as a therapeutic agent.

International Journal of Molecular SciencesVol. 26(24)
Russian Academy of Sciences (RU), St Petersburg University (RU), Lomonosov Moscow State University (RU), Sirius University of Science and Technology (RU), North-Caucasian Zonal Research Institute of Horticulture and Viticulture (RU), All-Russian Research Institute of Agricultural Microbiology (RU), Institute of Bioorganic Chemistry (RU), Institute of Molecular and Cell Biology (RU), Federal Research Centre «Fundamentals of Biotechnology» of the Russian Academy of Sciences (RU), Moscow State University (TJ)
Ministry of Science and Higher Education of the Russian Federation
Openalex Percentile: Top 65%
RNA and protein synthesis mechanisms
DOIOpen Access PDFOpenAlex
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