Abstract
A new strain of Bacillus velezensis NDB was isolated from Xiangshan Harbor and antibacterial test revealed antibacterial activity of this strain against 12 major pathogenic bacteria. The whole genome of the bacterium was sequenced and found to consist of a 4,214,838 bp circular chromosome and a 7410 bp circular plasmid. Furthermore, it was predicted by AntiSMASH and BAGEL4 to have 12 clusters of secondary metabolism genes for the synthesis of the inhibitors, fengycin, bacillomycin, macrolactin H, bacillaene, and difficidin, and there were also five clusters encoding potentially novel antimicrobial substances, as well as three bacteriocin biosynthesis gene clusters of amylocyclicin, ComX1, and LCI. qRT-PCR revealed significant up-regulation of antimicrobial secondary metabolite synthesis genes after 24 h of antagonism with pathogenic bacteria. Furthermore, MALDI-TOF mass spectrometry revealed that it can secrete surfactin non-ribosomal peptide synthase and polyketide synthase to exert antibacterial effects. GC-MS was used to analyze methanol extract of B. velezensis NDB, a total of 68 compounds were identified and these metabolites include 16 amino acids, 17 acids, 3 amines, 11 sugars, 11 alcohols, 1 ester, and 9 other compounds which can inhibit pathogenic bacteria by initiating the antibiotic secretion pathway. A comparative genomic analysis of gene families showed that the specificity of B. velezensis NDB was mainly reflected in environmental adaptability. Overall, this research on B. velezensis NDB provides the basis for elucidating its biocontrol effect and promotes its future application as a probiotic.
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Data availability
The sequences have been deposited sited in the NCBI Sequence Read Archive Database under the accession number CP135184-CP135185. The authors declare that [the/all other] data supporting the findings of this study are available within the paper (and its supplementary information files). The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Funding
This work was sponsored by the One Health Interdisciplinary Research Project, Ningbo University; the Ningbo Science and Technology Correspondent Team Project; the Natural Science Foundation of Zhejiang Province (LGC22C190002); and the KC Wong Magna Fund at Ningbo University.
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W.Z. performed the conceptualization, data curation, formal analysis, and writing-original draft; Z.W., W.Z., Z.Z., L.Y., L.S., W.Q., and E.S. performed validation and visualization; H.J., Z.J., X.J., and Y.X. performed supervision, validation, and visualization; and Z.Z. designed the experiments, contributed to experimental materials, writed the original draft and revised the paper; S.X., and W.R. designed the experiments, contributed to experimental materials, and revised the paper. All authors read and approved the final manuscript.
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Supplementary file1 Table S1 Antibacterial results of B. velezensis NDB strains screened by the disk-diffusion assay. Table S2 Prediction of secondary metabolites of B. velezensis NDB genome. Table S3 Prediction results of B. velezensis NDB bacteriocins. Supplementary Table 4 Primers used in this study. Table S5 Concentration of intracellular metabolites of B. velezensis NDB. (DOCX 34 KB)
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Wang, Z., Zhang, W., Wang, Z. et al. Analysis of antimicrobial biological activity of a marine Bacillus velezensis NDB. Arch Microbiol 206, 131 (2024). https://doi.org/10.1007/s00203-024-03861-4
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DOI: https://doi.org/10.1007/s00203-024-03861-4