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Plipastatin and surfactin coproduction by Bacillus subtilis pB2-L and their effects on microorganisms

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Abstract

To convert the lipopeptide non-producer strain Bacillus subtilis pB2 into a plipastatin and surfactin coproducer, a gene expression cassette composed of a constitutive promoter (P43), functional gene sfp, and pleiotropic regulatory gene degQ was integrated into the chromosomal amyE locus of strain B. subtilis pB2 by homologous recombination, which generated a plipastatin and surfactin co-producer. Thirteen plipastatins and fifteen surfactins were identified in lipopeptide extracts using analytical techniques, and their effects on microorganisms were described by microscopic, cytoskeleton analysis and flow-cytometry, respectively. Plipastatins isolated from the engineered strain pB2-L exhibited strong antifungal activity (MIC 16 μg ml−1) by disrupting the cell walls, membranes and cytoskeleton of Fusarium oxysporum f. sp. cucumerinum hyphae. Surfactins affected the cell membrane of Staphylococcus aureus (MIC 20 μg ml−1), resulting in nucleic acid leakage and ultimately, cell death. Based on the convenience of genetic manipulation in the engineering strain, this work could be useful for the rational design of lipopeptide synthetases via the recombination of gene fragments to generate arrays of peptide derivatives and thus expand the diversity of microbial-produced lipopeptides.

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Acknowledgements

This work was supported by Grants from the National Natural Science Foundation of China (Grant No. 31271828) and the National Science and Technology Support program (Grant No. 2011BAD23B05).

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Authors and Affiliations

  1. Key Laboratory of Food Processing and Quality Control, Ministry of Agriculture of China, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, People’s Republic of China

    Ling Gao, Jinzhi Han, Hongxia Liu, Xiaoxu Qu, Zhaoxin Lu & Xiaomei Bie

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  1. Ling Gao
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  2. Jinzhi Han
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  3. Hongxia Liu
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  4. Xiaoxu Qu
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  6. Xiaomei Bie
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Correspondence to Xiaomei Bie.

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Supplementary material 1 (TIFF 707 kb)

Fig. S1. LC–ESI–MS/MS of [M+H]+ ions at (a) 1491.84 m/z, plipastatin A (C18), (b) 1489.86 m/z, plipastatin A (unsaturated C18), (c) 1519.87 m/z, plipastatin A (C20)

Supplementary material 2 (TIFF 294 kb)

Fig. S2. LC–ESI–MS/MS of [M+H]+ ion at 1026.61 m/z representing linear surfactin

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Gao, L., Han, J., Liu, H. et al. Plipastatin and surfactin coproduction by Bacillus subtilis pB2-L and their effects on microorganisms. Antonie van Leeuwenhoek 110, 1007–1018 (2017). https://doi.org/10.1007/s10482-017-0874-y

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  • DOI: https://doi.org/10.1007/s10482-017-0874-y

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