Skip to main content
SpringerLink
Log in

Heterologous expression, purification, and phylogenetic analysis of oil-degrading biosurfactant biosynthesis genes from the marine sponge-associated Bacillus licheniformis NIOT-06

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

Surfactin is a lipopeptide, composed of one β-hydroxy fatty acid, a long fatty acid moiety, and seven amino acids. In this study, the biosurfactant biosynthesis genes; 4′-pantetheinyl transferase (sfp), phosphopantetheinyl transferase (sfpO), and surfactin synthetase (srfA) have been characterized from the marine sponge-associated Bacillus licheniformis NIOT-06 from the Andaman and Nicobar Islands. The purified recombinant biosurfactant revealed excellent emulsification activity with crude oil and kerosene. Reverse-phase high-performance liquid chromatography resolved the purified recombinant biosurfactant into several fractions and one of which had significant surface tension reducing property. Fourier transform infrared spectroscopy spectrum also revealed the presence of C–N–N, alkenes, and N–H as the functional groups, and a similar overlapping pattern was observed with that of standard lipopeptide surfactin. The diversity and phylogeny of sfp, sfpO, and srfA gene sequences were compared with other eubacteria. The sfp, sfpO, and srfA gene sequences obtained from Bacillus licheniformis NIOT-06 were diverse and appeared to be partially conserved when compared with the GenBank reported sequences of several eubacteria.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Rosenberg E, Ron EZ (1999) High and low molecular mass microbial surfactants. Appl Microbiol Biotechnol 52:154–162

    Article  CAS  Google Scholar 

  2. Arima K, Kakinuma A, Tamur G (1968) Surfactin, a crystalline peptide lipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochem Biophys Res Commun 31:488–494

    Article  CAS  Google Scholar 

  3. Donio MBS, Ronica FA, Thanga Viji V, Velmurugan S, Adlin Jenifer J, Michaelbabu M, Dhar P, Citarasu T (2013) Halomonas sp. BS4, a biosurfactant producing halophilic bacterium isolated from solar salt works in India and their biomedical importance. Springer Plus 2:149

    Article  Google Scholar 

  4. Anburajan L, Meena B, Joseph TC, Dheenan PS, Vinithkumar NV, Dharani G, Kirubagaran R (2014) Functional and molecular characterization of a lipopeptide surfactant from marine sponge-associated eubacteria Bacillus licheniformis NIOT-AMKV06 of Andaman and Nicobar Islands, India. Mar Pollut Bull 82:76–85

    Article  Google Scholar 

  5. Bewley CA, Holland NA, Faulkner DJ (1996) Two classes of metabolites from Theonella swinhoei are localized in distinct populations of bacterial symbionts. Experientia 52:71–75

    Article  Google Scholar 

  6. Kennedy J, Marchesi JR, Dobson ADW (2007) Metagenomic approaches to exploit the biotechnological potential of the microbial consortia of marine sponges. Appl Microbiol Biotechnol 75:11–20

    Article  CAS  Google Scholar 

  7. Ito S, Kobayashi T, Ara K, Ozaki K, Kawai S, Hatada Y (1998) Alkaline detergent enzymes from alkaliphiles: enzymatic properties, genetics, and structures. Extremophiles 2:185–190

    Article  CAS  Google Scholar 

  8. Salonen MSS, Vuorio R, Andersson MA, Kampfer P, Andersson MC, Buzalski TH, Scoging AC (1999) Toxigenic strains of Bacillus licheniformis related to food poisioning. Appl Environ Microbiol 65:4637–4645

    Google Scholar 

  9. Ausubel FM, Brent R, Kingston RE, Moore DD, Scidman JG, Smith JA, Strichi K (1994) Current protocols in molecular biology 2.0.1–2.14.8

  10. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature 227:680–685

    Article  CAS  Google Scholar 

  11. Ono H, Sawada K, Khunajakr N, Tao T, Yamamoto M, Hiramoto M, Shinmyo A, Takano M, Murooka Y (1999) Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacterium, Halomonas elongata. J Bacteriol 181:91–99

    CAS  Google Scholar 

  12. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Ann Biochem 72:248–254

    Article  CAS  Google Scholar 

  13. Cirigliano MC, Carman GM (1985) Purification and characterization of liposan, a bioemulsifier from Candida lipolytica. Appl Environ Microbiol 50:846–850

    CAS  Google Scholar 

  14. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetic analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  Google Scholar 

  15. Lee SC, Lee SJ, Kim SH, Park IH, Lee YS, Chung SY, Choi YL (2008) Characterization of new biosurfactant produced by Klebsiella sp. Y6-1 isolated from waste soybean oil. Bioresour Technol 99:2288–2292

    Article  CAS  Google Scholar 

  16. Koumoutsi A, Chen XH, Henne A, Liesegang H, Hitzeroth G, Franke P, Vater J, Borriss R (2004) Structural and functional characterization of gene clusters directing nonribosomal synthesis of bioactive cyclic lipopeptides in Bacillus amyloliquefaciens strain FZB42. J Bacteriol 186:1084–1096

    Article  CAS  Google Scholar 

  17. Nakano MN, 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

    CAS  Google Scholar 

  18. Porob S, Nayak S, Fernandes A, Padmanabhan P, Patil BA, Meena RM, Ramaiah N (2013) PCR screening for the surfactin (sfp) gene in marine Bacillus strains and its molecular characterization from Bacillus tequilensis NIOS11. Turkish J Biol 37:212–221

    CAS  Google Scholar 

  19. Kiran SG, Thomas TA, Selvin J, Sabarathnam B, Lipton AP (2010) Optimization and characterization of a new lipopeptide biosurfactant produced by marine Brevibacterium aureum MSA13 in solid state culture. Bioresour Technol 101:2389–2396

    Article  Google Scholar 

  20. Sausa T, Bhosle S (2012) Isolation and characterization of a lipopeptide bioemulsifier produced by Pseudomonas nitroreducens TSBMJ10 isolated from a mangrove ecosystem. Bioresour Technol 123:256–262

    Article  Google Scholar 

  21. Joshi S, Bharucha C, Desai AJ (2008) Production of biosurfactant and antifungal compound by fermented food isolate Bacillus subtilis 20B. Bioresour Technol 99:4603–4608

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the financial support given by the Earth System Sciences Organization (ESSO), Ministry of Earth Sciences (MoES), Government of India, New Delhi to conduct the research. The authors are thankful to Dr. M. A. Atmanand, Director, ESSO-National Institute of Ocean Technology (ESSO-NIOT), MoES, Chennai, for constant support and encouragement to perform this research. The authors are profoundly thankful to Dr. M. Vijayakumaran for critical comments and suggestions to improve this manuscript.

Author information

Author notes

    Authors and Affiliations

    1. Andaman and Nicobar Centre for Ocean Science and Technology, Earth System Sciences Organization-National Institute of Ocean Technology (ESSO-NIOT), Dollygunj P.O., Port Blair, 744103, Andaman and Nicobar Islands, India

      Lawrance Anburajan, Balakrishnan Meena, Nambali Valsalan Vinithkumar & Palaiya Sukumaran Dheenan

    2. Marine Biotechnology Division, Ocean Science and Technology for Islands Group, ESSO-NIOT, Ministry of Earth Sciences, Government of India, Chennai, 600100, Tamil Nadu, India

      Rangamaran Vijaya Raghavan, Divya Shridhar, Gopal Dharani & Ramalingam Kirubagaran

    3. Microbiology, Fermentation and Biotechnology Division, Central Institute of Fisheries Technology, Indian Council of Agricultural Research, Ministry of Agriculture, Government of India, Kochi, 682029, Kerala, India

      Toms Cheriath Joseph

    Authors
    1. Lawrance Anburajan
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Balakrishnan Meena
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Rangamaran Vijaya Raghavan
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Divya Shridhar
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Toms Cheriath Joseph
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Nambali Valsalan Vinithkumar
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Gopal Dharani
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Palaiya Sukumaran Dheenan
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Ramalingam Kirubagaran
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding authors

    Correspondence to Lawrance Anburajan or Ramalingam Kirubagaran.

    Additional information

    L. Anburajan and B. Meena contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Anburajan, L., Meena, B., Raghavan, R.V. et al. Heterologous expression, purification, and phylogenetic analysis of oil-degrading biosurfactant biosynthesis genes from the marine sponge-associated Bacillus licheniformis NIOT-06. Bioprocess Biosyst Eng 38, 1009–1018 (2015). https://doi.org/10.1007/s00449-015-1359-x

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00449-015-1359-x

    Keywords

    Search

    Navigation