Skip to main content
SpringerLink
Log in

Expression of sfp gene and hydrocarbon degradation by Bacillus subtilis

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Bacillus subtilis C9 produces a lipopeptide-type biosurfactant, surfactin, and rapidly degrades alkanes up to a chain length of C19. The nucleotide sequence of the sfp gene cloned from B. subtilis C9 was determined and its deduced amino acid sequence showed 100% homology with the sfp gene reported before [Nakano et al. (1992) Mol. Gen. Genet. 232: 313–321]. To transform a non-surfactin producer, B. subtilis 168, to a surfactin producer, the sfp gene cloned from B. subtilis C9 was expressed in B. subtilis 168. The transformed B. subtilis SB103 derivative of the strain 168 was shown to produce surfactin measured by its decrease in surface tension, emulsification activity, and TLC analysis of the surface active compound isolated from the culture broth. Like B. subtilis C9, B. subtilis SB103 containing sfp gene readily degraded aliphatic hydrocarbons (C10−19), though its original strain did not. The addition of surfactin (0.5%, w/v) to the culture of B. subtilis 168 significantly stimulated the biodegradation of hydrocarbons of the chain lengths of 10–19; over 98% of the hydrocarbons tested were degraded within 24 h of incubation. These results indicate that the lipopeptide-type biosurfactant, surfactin produced from B. subtilis enhances the bioavailability of hydrophobic hydrocarbons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anagnostopoulos C, Spizizen J (1961) Requirements for transformation in Bacillus subtilis. J. Bacteriol. 81: 741–746.

    Google Scholar 

  • Andreas KK, Kappeli O, Fiechter A, Reiser J (1991) Hydrocarbon assimilation and biosurfactant production in Pseudomonas aeruginosa mutants. J. Bacteriol. 173: 4212–4219.

    PubMed  Google Scholar 

  • Banat IM (1995) Characterization of biosurfactants and their use in pollution removal-State of the art. Acta Biotechnol. 15: 251–267.

    Google Scholar 

  • Cosmina P, Rodriguez F, de Ferra F, Grandi G, Perego M, Venema G, van Sinderen D (1993) Sequence and analysis of the genetic locus responsible for surfactin synthesis in Bacillus subtilis. Mol. Microbiol. 8: 821–831.

    PubMed  Google Scholar 

  • Desai JD (1987) Microbial surfactants: Evaluation, types, production and future applications. J. Sci. Ind. Res. 46: 440–449.

    Google Scholar 

  • Doi RH (1983) Reading, In: Rodriguez RL, Tait RC, eds. Recombinant DNA Techniques. Reading, MA: Addison Wesley, pp. 161–163.

    Google Scholar 

  • Georgiou G, Lin SC, Sharma MM(1992) Surface-active compounds from microorganisms. Bio/Technology 10: 60–65.

    PubMed  Google Scholar 

  • Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of Escherichia coli with plasmids. Gene 96: 23–28.

    PubMed  Google Scholar 

  • Jain DK, Collins-Thompson DL, Lee H, Trevors JT (1991) A drop-collapsing test for screening surfactant-producing microorganisms. J. Microbiol. Meth. 13: 271–279.

    Google Scholar 

  • Kim HS, Lee CH, Suh HH, Ahn KH, Oh HM, Kwon GS, Yang JW, Yoon BD (1997a) A lipopeptide biiosurfactant produced by Bacillus subtilis C9 selected through the oil film-collapsing assay. J. Microbiol. Biotechnol. 7: 180–188.

    Google Scholar 

  • Kim HS, Yoon BD, Lee CH, Suh HH, Oh HM, Katsuragi T, Tani Y (1997b) Production and properties of a lipopeptide biosurfactant from Bacillus subtilis C9. J. Ferment. Bioengin. 84: 41–46.

    Google Scholar 

  • Koch AK, Käppeli O, Fiechter A, Reiser J (1991) Hydrocarbon assimilation and biosurfactant production in Pseudomonas aeruginosa mutants. J. Bacteriol. 173: 4212–4219.

    PubMed  Google Scholar 

  • Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

    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.

    PubMed  Google Scholar 

  • Nakano MM, Corbell N, Besson J, Zuber P (1992) Isolation and characterization of sfp: a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis. Mol Gen. Genet. 232: 313–321.

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • Quadri LEN, Weinreb PH, Lei M, Nakano MM, Zuber P, Walsh CT (1998) Characterization of Sfp, a Bacillus subtilis phosphopantetheinyl transferase for peptidyl carrier protein domains in peptide synthetases. Biochemistry 37: 1585–1595.

    PubMed  Google Scholar 

  • Reuter K, Mofid MR, Marahiel MA, Ficner R (1999) Crystal structure of the surfactin synthetase-activating enzyme Sfp: a prototype of the 4′-phosphopantetheinyl transferase superfamily. EMBO J. 18: 6823–6831.

    PubMed  Google Scholar 

  • Rosenberg E, Zuckerberg A, Rubinovitz C, Gutnick DL (1979) Emulsifier of Arthrobacter sp. RAG-1: isolation and emulsifying properties. Appl. Environ. Microbiol. 37: 402–408.

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • Spizizen J (1958) Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc. Natl. Acad. Sci. 44: 1072–1078.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Environmental Bioresources Laboratory, Korea

    Hee-Sik Kim, Hee-Mock Oh & Yong-Il Park

  2. Microbial and Bioprocess Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology, Oeun-dong 52, Yusong, Taejon , 305-333, Korea

    Seong-Bin Kim, Seung-Hwan Park & Byung-Dae Yoon

  3. Department of Microbiology, Chungbuk National University, Cheong-ju , 361-763, Korea

    Chi-Kyung Kim

  4. Laboratory of Cell Biotechnology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara , 630-01, Japan

    Tohoru Katsuragi & Yoshiki Tani

Authors
  1. Hee-Sik Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Seong-Bin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Seung-Hwan Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hee-Mock Oh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yong-Il Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chi-Kyung Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tohoru Katsuragi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yoshiki Tani
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Byung-Dae Yoon
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, HS., Kim, SB., Park, SH. et al. Expression of sfp gene and hydrocarbon degradation by Bacillus subtilis. Biotechnology Letters 22, 1431–1436 (2000). https://doi.org/10.1023/A:1005675231949

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005675231949

Search

Navigation