Abstract
Rhodococcus erythropolis S43 is an arsenic-tolerant actinobacterium isolated from an arsenic contaminated soil. It has been shown to produce siderophores when exposed to iron-depleting conditions. In this work, strain S43 was shown to have the putative heterobactin production cluster htbABCDEFGHIJ(K). To induce siderophore production, the strain was cultured in iron-depleted medium in presence and absence of sodium arsenite. The metabolites produced by S43 in the colorimetric CAS and As-mCAS assays, respectively, showed iron- and arsenic-binding properties reaching a chelating activity equivalent to 1.6 mM of desferroxamine B in the supernatant of the culture without arsenite. By solid-phase extraction and two subsequent HPLC separations from both cultures, several fractions were obtained, which contained CAS and As-mCAS activity and which were submitted to LC-MS analyses including fragmentation of the major peaks. The mixed-type siderophore heterobactin B occurred in all analyzed fractions, and the mass of the “Carrano heterobactin A” was detected as well. In addition, generation of a molecular network based on fragment spectra revealed the occurrence of several other compounds with heterobactin-like structures, among them a heterobactin B variant with an additional CH2O moiety. 1H NMR analyses obtained for preparations from the first HPLC step showed signals of heterobactin B and of “Carrano heterobactin A” with different relative amounts in all three samples. In summary, our results reveal that in R. erythropolis S43, a pool of heterobactin variants is responsible for the iron- and arsenic-binding activities.
Key points
• Several heterobactin variants are the arsenic-binding compounds in Rhodococcus erythropolis S43.
• Heterobactin B and the compound designated heterobactin A by Carrano are of importance.
• In addition, other heterobactins with ornithine in the backbone exist, e.g., the new heterobactin C.
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Acknowledgements
The authors thank Thomas Heine for providing access to the draft genome sequence of R. erythropolis S43 on the RAST server.
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The authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files.
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This work was supported by project Fondecyt #1170799 from the government of Chile, and USA1799 and Dicyt-USACH from University of Santiago. GR-M has received a doctoral fellowship from CONICYT (COD: 21140962). A stay of GR-M in Freiberg was supported by the DAAD project YoungGEOMATENUM-International. The work of DT and MSt was supported by BMBF junior research group BakSolEx (#033R147). JEB acknowledges funding from the German Research Foundation (BA 4193/6-1) and thanks the German federal state of North Rhine-Westphalia for funding the mass spectrometer (Forschungsgroßgeräte der Länder).
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MSc and GL conceived and designed research. GR-M, MSe, MSt conducted experiments. JEB, DT, AO, and BM contributed new reagents or analytical tools. GR-M and CHRS analyzed data. GR-T, GL, and MSc wrote the manuscript. All authors read and approved the manuscript.
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Retamal-Morales, G., Senges, C.H.R., Stapf, M. et al. Isolation and characterization of arsenic-binding siderophores from Rhodococcus erythropolis S43: role of heterobactin B and other heterobactin variants. Appl Microbiol Biotechnol 105, 1731–1744 (2021). https://doi.org/10.1007/s00253-021-11123-2
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DOI: https://doi.org/10.1007/s00253-021-11123-2