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Isolation and characterization of sfp: a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis

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Summary

The sfp gene is required for cells of Bacillus subtilis to become producers of the lipopeptide antibiotic surfactin. sfp was isolated and its nucleotide sequence was determined. sfp was expressed in Escherichia coli and its putative product was purified for use in antibody production and in amino acid sequence analysis. The gene was plasmid-amplified in B. subtilis, where it conferred a Srf+ phenotype on sfp 0 (surfactin non-producing) cells. Overproduction of Sfp in B. subtilis did not cause production of an increased amount of surfactin and resulted in the repression of a lacZ transcriptional fusion of the srfA operon, which encodes enzymes that catalyze surfactin synthesis. We propose that sfp represents an essential component of peptide synthesis systems and also plays a role, either directly or indirectly, in the regulation of surfactin biosynthesis gene expression.

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References

  • Arima K, Kakinuma A, Tamura G (1968) Surfactin, a crystalline peptidelipid surfactant produced by Bacillus subtilis: Isolation, characterization and its inhibition of fibrin clot formation. Biochem Biophys Res Commun 31:488–494

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology. Greene Publishing Associates and Wiley-Interscience, NY

    Google Scholar 

  • Dubnau D (1989) The competence regulon of Bacillus subtilis. In: Smith I, Slepecky RA, Setlow P (eds) Regulation of procaryotic development: A structural and functional analysis of bacterial sporulation and germination. American Society for Microbiology, Washington, DC, pp 147–166

    Google Scholar 

  • Dubnau D (1991) The regulation of genetic competence in Bacillus subtilis. Mol Microbiol 5:11–18

    Google Scholar 

  • Dubnau D, Davidoff-Abelson R (1971) Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol 56:209–221

    Google Scholar 

  • Freese E, Heinze J (1983) Metabolic and genetic control of bacterial sporulation. In: Hurst A, Gould GW (eds) The bacterial spore, vol 2. Academic Press, London, pp 101–172

    Google Scholar 

  • Grundy FJ, Henkin TM (1990) Cloning and analysis of the Bacillus subtilis rpsD gene, encoding ribosomal protein S4. J Bacteriol 172:6372–6379

    Google Scholar 

  • Gryczan T, Shivakumar AG, Dubnau D (1980) Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis. J Bacteriol 141:246–253

    Google Scholar 

  • Harlow E, Lane D (1988) Antibodies. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 313–315

    Google Scholar 

  • Herskowitz I (1989) A regulatory hierarchy for cell specialization in yeast. Nature 342:749–757

    Google Scholar 

  • Hoch JA, Barat M, Anagnostopoulos C (1967) Transformation and transduction in recombination-defective mutants of Bacillus subtilis. J Bacteriol 93:1925–1937

    Google Scholar 

  • Igo MM, Losick R (1986) Regulation of a promoter that is utilized by minor forms of RNA polymerase holoenzyme in Bacillus subtilis. J Mol Biol 191:615–624

    Google Scholar 

  • Katz E, Demain AL (1977) The peptide antibiotics of Bacillus: chemistry, biogenesis, and possible functions. Bacteriol Rev 41:449–474

    Google Scholar 

  • Kleinkauf H, von Dohren H (1987) Biosynthesis of peptide antibiotics. Annu Rev Microbiol 41:259–289

    Google Scholar 

  • Kratzschmar JM, Krause M, Marahiel MA (1989) Gramicidin S biosynthesis operon containing the structural genes grsA and grsB has an open reading frame encoding a protein homologous to fatty acid thioesterases. J Bacteriol 171:5422–5429

    Google Scholar 

  • Kurahashi K (1974) Biosynthesis of small peptides. Annu Rev Biochem 43:445–459

    Google Scholar 

  • Kurjan J, Whiteway M (1990) The Saccharomyces cerevisiae pheromone response pathway. Semin Dev Biol 1:151–158

    Google Scholar 

  • Lipmann F (1980) Bacterial production of antibiotic polypeptides by thiol-linked synthesis on protein template. Adv Microbiol Physiol 21:227–260

    Google Scholar 

  • Love PE, Lyle MJ, Yasbin RE (1985) DNA-damage-inducible (din) loci are transcriptionally activated in competent Bacillus subtilis. Proc Natl Acad Sci USA 82:6201–6205

    Google Scholar 

  • Martin IF, Liras P (1989) Organization and expression of genes involved in the biosynthesis of antibiotics and other secondary metabolites. Annu Rev Microbiol 43:173–206

    Google Scholar 

  • Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78

    Google Scholar 

  • Monaghan RL, Tkacz JS (1990) Bioactive microbial products: focus upon mechanism of action. Annu Rev Microbiol 44:271–301

    Google Scholar 

  • Nakano MM, Zuber P (1989) Cloning and characterization of srfB, a regulatory gene involved in surfactin production and competence in Bacillus subtilis. J Bacteriol 171:5347–5353

    Google Scholar 

  • Nakano MM, Zuber P (1990a) Molecular biology of antibiotic production in Bacillus. CRC Crit Rev Biotech 10:223–240

    Google Scholar 

  • Nakano MM, Zuber P (1990b) Identification of genes required for the biosynthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis. In: Hoch J, Ganesan AT (eds) Genetics and biotechnology of bacilli, vol 3. Academic Press, New York, pp 397–405

    Google Scholar 

  • Nakano MM, Marahiel MA, Zuber P (1988) Identification of a genetic locus required for biosynthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis. J Bacteriol 170:5662–5668

    Google Scholar 

  • Nakano MM, Magnuson R, Myers A, Curry J, Grossman AD, Zuber P (1991a) srfA is an operon required for surfactin production, competence development, and efficient sporulation in Bacillus subtilis. J Bacteriol 173:1770–1778

    Google Scholar 

  • Nakano MM, Xia L, Zuber P (1991b) Transcription initiation region of the srfA operon which is controlled by the ComP-ComA signal transduction system in Bacillus subtilis. J Bacteriol 173:5487–5493

    Google Scholar 

  • Niaudet B, Ehrlich SD (1979) In vitro genetic labeling of Bacillus subtilis cryptic plasmid pHV400. Plasmid 2:48–58

    Google Scholar 

  • Sanger R, Nicklen W, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • van Sinderen D, Withoff S, Boels H, Venema G (1990) Isolation and characterization of comL, a transcription unit involved in competence development of Bacillus subtilis. Mol Gen Genet 224:396–404

    Google Scholar 

  • Vater J (1989) Lipopeptides, an interesting class of microbial secondary metabolites. In: Schlunegger UP (ed) Biologically active molecules. Springer-Verlag, Berlin Heidelberg, pp 27–37

    Google Scholar 

  • Vining LC (1990) Functions of secondary metabolites. Annu Rev Microbiol 44:395–427

    Google Scholar 

  • Weinrauch Y, Guillen N, Dubnau DA (1989). Sequence and transcription mapping of Bacillus subtilis competence genes comB and comA, one of which is related to a family of bacterial regulatory determinants. J Bacteriol 171:5362–5375

    Google Scholar 

  • Weinrauch Y, Penchev R, Dubnau E, Smith I, Dubnau D (1990) A Bacillus subtilis regulatory gene product for genetic competence and sporulation resembles sensor protein members of the bacterial two-component signal-transduction systems. Genes Dev 4:860–872

    Google Scholar 

  • Youngman P, Perkins JB, Losick R (1984) A novel method for the rapid cloning in Escherichia coli of Bacillus subtilis chromosomal DNA adjacent to Tn917 insertions. Mol Gen Genet 195:424–433

    Google Scholar 

  • Zuber P (1985) Localizing the site of spoO-dependent regulation in the spoVG promoter of Bacillus subtilis. In: Hoch JA, Setlow P (eds) Spores IX. American Society for Microbiology, Washington, DC, pp 149–156

    Google Scholar 

  • Zuber P, Losick R (1987) Role of AbrB in SpoOA- and SpoOB-dependent utilization of a sporulation promoter in Bacillus subtilis. J Bacteriol 169:2223–2230[ba/]

    Google Scholar 

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Communicated by T. Lengeler

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Nakano, M.M., Corbell, N., Besson, J. et al. Isolation and characterization of sfp: a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis . Molec. Gen. Genet. 232, 313–321 (1992). https://doi.org/10.1007/BF00280011

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