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New approach for the detection of non-ribosomal peptide synthetase genes in Bacillus strains by polymerase chain reaction

  • Applied Genetics and Molecular Biotechnology
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Abstract

Bacillus strains produce non-ribosomal lipopeptides that can be grouped into three families: surfactins or lichenysins, iturins and fengycins or plispastatins. These biosurfactants show a broad spectrum of biological activities. To detect strains able to produce these lipopeptides, a new polymerase chain reaction screening approach was developed using degenerated primers based on the intraoperon alignment of adenylation and thiolation nucleic acid domains of all enzymes implicated in the biosynthesis of each lipopeptide family. The comparative bioinformatics analyses of each operon led to the design of four primer pairs for the three families taking into account the differences between open reading frames of each synthetase gene. Tested on different Bacillus sp. strains, this technique was used successfully to detect not only the expected genes in the lipopeptide producing strains but also the presence of a plispastatin gene in Bacillus subtilis ATCC 21332 and a gene showing a high similarity with the polyketide synthase type I gene in the B. subtilis ATCC 6633 genome. It also led to the discovery of the presence of non-ribosomal peptide synthetase genes in Bacillus thuringiensis serovar berliner 1915 and in Bacillus cereus LMG 2098. In addition, this work highlighted the differences between the fengycin and plipastatin operon at DNA level.

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Acknowledgements

This work was supported by the Université des Sciences et Technologies de Lille, the Region Nord Pas de Calais, the Ministere de la Recherche Scientifique (ANR) and the European Funds for Regional Development. Arthur Tapi has a fellowship from Ivorian government. The authors thank William Everett for the re-reading of the manuscript.

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Authors and Affiliations

  1. Laboratoire des Procédés Biologiques, Génie Enzymatique et Microbien, UPRES-EA 1026, Polytech’Lille/IUTA, Université des Sciences et Technologies de Lille, Av. Paul Langevin, Villeneuve d’Ascq, 59655, Lille, France

    Arthur Tapi, Marlène Chollet-Imbert & Philippe Jacques

  2. Institut de Recherches Microbiologiques JM Wiame, Av. E. Gryson 1, 1070, Brussels, Belgium

    Bart Scherens

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  1. Arthur Tapi
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  2. Marlène Chollet-Imbert
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  4. Philippe Jacques
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Correspondence to Philippe Jacques.

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Table 1

Number and size (*bp) of amplicons obtained by in silico PCR (AmplifX Software) from lipopeptide genes deduced from the genome of strains indicated in Table 3. (DOC 33 kb)

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Tapi, A., Chollet-Imbert, M., Scherens, B. et al. New approach for the detection of non-ribosomal peptide synthetase genes in Bacillus strains by polymerase chain reaction. Appl Microbiol Biotechnol 85, 1521–1531 (2010). https://doi.org/10.1007/s00253-009-2176-4

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