Purification and characterization of a surfactin-like molecule produced by Bacillus sp. H2O-1 and its antagonistic effect against sulfate reducing bacteria

BMC Microbiol. 2012 Nov 7:12:252. doi: 10.1186/1471-2180-12-252.

Abstract

Background: Bacillus sp. H2O-1, isolated from the connate water of a Brazilian reservoir, produces an antimicrobial substance (denoted as AMS H2O-1) that is active against sulfate reducing bacteria, which are the major bacterial group responsible for biogenic souring and biocorrosion in petroleum reservoirs. Thus, the use of AMS H2O-1 for sulfate reducing bacteria control in the petroleum industry is a promising alternative to chemical biocides. However, prior to the large-scale production of AMS H2O-1 for industrial applications, its chemical structure must be elucidated. This study also analyzed the changes in the wetting properties of different surfaces conditioned with AMS H2O-1 and demonstrated the effect of AMS H2O-1 on sulfate reducing bacteria cells.

Results: A lipopeptide mixture from AMS H2O-1 was partially purified on a silica gel column and identified via mass spectrometry (ESI-MS). It comprises four major components that range in size from 1007 to 1049 Da. The lipid moiety contains linear and branched β-hydroxy fatty acids that range in length from C13 to C16. The peptide moiety contains seven amino acids identified as Glu-Leu-Leu-Val-Asp-Leu-Leu.Transmission electron microscopy revealed cell membrane alteration of sulfate reducing bacteria after AMS H2O-1 treatment at the minimum inhibitory concentration (5 μg/ml). Cytoplasmic electron dense inclusions were observed in treated cells but not in untreated cells. AMS H2O-1 enhanced the osmosis of sulfate reducing bacteria cells and caused the leakage of the intracellular contents. In addition, contact angle measurements indicated that different surfaces conditioned by AMS H2O-1 were less hydrophobic and more electron-donor than untreated surfaces.

Conclusion: AMS H2O-1 is a mixture of four surfactin-like homologues, and its biocidal activity and surfactant properties suggest that this compound may be a good candidate for sulfate reducing bacteria control. Thus, it is a potential alternative to the chemical biocides or surface coating agents currently used to prevent SRB growth in petroleum industries.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification
  • Anti-Bacterial Agents / metabolism*
  • Bacillus / genetics*
  • Bacillus / isolation & purification
  • Bacillus / metabolism*
  • Brazil
  • Cell Membrane / ultrastructure
  • Chromatography
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Lipopeptides / chemistry
  • Lipopeptides / genetics*
  • Lipopeptides / isolation & purification
  • Lipopeptides / metabolism*
  • Mass Spectrometry
  • Microbial Sensitivity Tests
  • Microscopy, Electron, Transmission
  • Molecular Sequence Data
  • Molecular Weight
  • Sequence Analysis, DNA
  • Sulfur-Reducing Bacteria / drug effects*
  • Sulfur-Reducing Bacteria / ultrastructure
  • Surface-Active Agents / chemistry
  • Surface-Active Agents / isolation & purification
  • Surface-Active Agents / metabolism*
  • Water Microbiology

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Lipopeptides
  • Surface-Active Agents

Associated data

  • GENBANK/JX575798