Abstract
The aim of this work was to develop a comparative study between Bacillus paralicheniformis TB197 and B. subtilis ATCC 21332 strains in terms of growth, cyclic lipopeptide production, nematicidal activity, and active lipopeptide characteristics. Crude lipopeptide extracts (CLEs) from their fermentation broths were obtained, and their nematicidal activity (NA) was estimated as the mean lethal dose (LD50), employing Caenorhabditis elegans. Using a bioguided approach, CLE components were fractionated by semipreparative thin layer chromatography, and active lipopeptides were characterized by mass spectrometry. Both strains produced similar concentrations of CLEs (p ≥ 0.05) (0.99 ± 0.11 and 1.14 ± 0.15 mg/mL by TB197 and ATCC 21332, respectively). The estimated LD50 values of CLEs from the TB197 and ATCC 21332 strains were 3.88 and 8.15 mg/mL, respectively, showing that the NA of the TB197 strain CLE was 2.1-fold higher (p ≤ 0.05). Mass spectrometry revealed that strain TB197 synthesizes several families of lipopeptides, namely, fengycin A (C14–C17), fengycin B (C16–C17), surfactin (C15–C17), and lichenysin (C12, C13, C14, and C16), from which fengycins and lichenysins possess the highest NA (100 and 60% mortality in C. elegans larvae, respectively), while the ATCC 21332 strain produces mainly surfactin (C13–C17) (NA 63% mortality). The main differences found in this study were that the TB197 strain has a higher tolerance to inhibition by the product, and the lipopeptides they synthesize have a higher nematicidal activity due to the diversity of families compared to ATCC 21332. Likewise, it was shown that more polar lipopeptides (fengycins) are more effective at causing mortality in C. elegans larvae.
Key points
• The nematicidal activity of lipopeptides from TB197 is higher than from ATCC 21332
• TB197 produces surfactin, lichenysin, and fengycin, while ATCC 21332 mainly produces surfactin
• The most polar lipopeptides (fengycins) cause more mortality in C. elegans L2
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Author Estefany Chavarria acknowledges the scholarship (number 867046) granted by the National Council on Science and Technology (CONACYT) for PhD studies. We are grateful to Innovak Global for the financial support and access to their R&D laboratories.
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All authors contributed to the study conception and design. EC performed all the experiments, analyzed the data, and wrote the original manuscript. EC and MG analyzed the mass spectra. AA supervised the research and reviewed the manuscript. All the authors read and approved the manuscript.
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Chavarria-Quicaño, E., De la Torre-González, F., González-Riojas, M. et al. Nematicidal lipopeptides from Bacillus paralicheniformis and Bacillus subtilis: A comparative study. Appl Microbiol Biotechnol 107, 1537–1549 (2023). https://doi.org/10.1007/s00253-023-12391-w
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DOI: https://doi.org/10.1007/s00253-023-12391-w