Applied Microbiology and Biotechnology

, Volume 64, Issue 2, pp 275–283 | Cite as

Inhibiting mild steel corrosion from sulfate-reducing bacteria using antimicrobial-producing biofilms in Three-Mile-Island process water

  • R. Zuo
  • D. Örnek
  • B. C. Syrett
  • R. M. Green
  • C.-H. Hsu
  • F. B. Mansfeld
  • T. K. Wood
Original Paper


Biofilms were used to produce gramicidin S (a cyclic decapeptide) to inhibit corrosion-causing, sulfate-reducing bacteria (SRB). In laboratory studies these biofilms protected mild steel 1010 continuously from corrosion in the aggressive, cooling service water of the AmerGen Three-Mile-Island (TMI) nuclear plant, which was augmented with reference SRB. The growth of both reference SRB (Gram-positive Desulfosporosinus orientis and Gram-negative Desulfovibrio vulgaris) was shown to be inhibited by supernatants of the gramicidin-S-producing bacteria as well as by purified gramicidin S. Electrochemical impedance spectroscopy and mass loss measurements showed that the protective biofilms decreased the corrosion rate of mild steel by 2- to 10-fold when challenged with the natural SRB of the TMI process water supplemented with D. orientis or D. vulgaris. The relative corrosion inhibition efficiency was 50–90% in continuous reactors, compared to a biofilm control which did not produce the antimicrobial gramicidin S. Scanning electron microscope and reactor images also revealed that SRB attack was thwarted by protective biofilms that secrete gramicidin S. A consortium of beneficial bacteria (GGPST consortium, producing gramicidin S and other antimicrobials) also protected the mild steel.


  1. Anonymous (1998) Multiple-tube fermentation technique for members of the coliform group. In: Greenberg AE, Clesceri LS, Eaton AD (eds) Standard methods for the examination of water and wastewater. American Public Health Association, American Water Works Association, and Water Environment Federation, New York, 9-45–9-51Google Scholar
  2. Azuma T, Harrison GI, Demain AL (1992) Isolation of a gramicidin S hyperproducing strain of B. brevis by use of a fluorescence activated cell sorting system. Appl Microbiol Biotechnol 38:173–178PubMedGoogle Scholar
  3. Beloglazov SM, Dzhafarov ZI, Polyakov VN, Demushin NN (1991) Quaternary ammonium salts as corrosion inhibitors of steel in the presence of sulfate-reducing bacteria. Prot Met (USSR) 27:810–813Google Scholar
  4. Borenstein SW (1994) Microbiologically influenced corrosion handbook. Woodhead, Cambridge, EnglandGoogle Scholar
  5. Chen M, Nagarajan V (1993) The role of signal peptide and mature protein in RNase (barnase) export from Bacillus subtilis Mol Gen Genet 239:409–415Google Scholar
  6. Cord-Ruwisch R, Kleinitz W, Widdel F (1987) Sulfate-reducing bacteria and their activities in oil production. J Petrol Technol 39:97–105Google Scholar
  7. Costerton JW, Geesey GG, Jones PA (1988) Bacterial biofilms in relation to internal corrosion monitoring and biocide strategies. Mater Perform 27:49–53Google Scholar
  8. Fontana MG (1986) Corrosion engineering, 3rd edn. McGraw-Hill, New YorkGoogle Scholar
  9. Franklin MJ, Nivens DE, Vass AA, Mittelman MW, Jack RF, Dowling NJE, White DC (1991) Effect of chlorine and chlorine/bromine biocide treatments on the number and activity of biofilm bacteria and on carbon steel corrosion. Corrosion 47:128–134Google Scholar
  10. Friedrich CL, Moyles D, Beveridge TJ, Hancock REW (2000) Antibacterial action of structurally diverse cationic peptides on Gram-positive bacteria. Antimicrob Agents Chemother 44:2086–2092PubMedGoogle Scholar
  11. Hamilton WA (1985) Sulphate-reducing bacteria and anaerobic corrosion. Annu Rev Microbiol 39:195–217CrossRefPubMedGoogle Scholar
  12. Jayaraman A, Cheng ET, Earthman JC, Wood TK (1997a) Axenic aerobic biofilms inhibit corrosion of SAE 1018 steel through oxygen depletion. Appl Microbiol Biotechnol 48:11–17PubMedGoogle Scholar
  13. Jayaraman A, Cheng ET, Earthman JC, Wood TK (1997b) Importance of biofilm formation for corrosion inhibition of SAE 1018 steel by axenic aerobic biofilms. J Ind Microbiol Biotechnol 18:396–401CrossRefPubMedGoogle Scholar
  14. Jayaraman A, Hallock PJ, Carson RM, Lee C-C, Mansfeld FB, Wood TK (1999a) Inhibiting sulfate-reducing bacteria in biofilms on steel with antimicrobial peptides generated in situ. Appl Microbiol Biotechnol 52:267–275CrossRefPubMedGoogle Scholar
  15. Jayaraman A, Mansfeld FB, Wood TK (1999b) Inhibiting sulfate-reducing bacteria in biofilms by expressing the antimicrobial peptides indolicidin and bactenecin. J Ind Microbiol Biotechnol 22:167–175CrossRefGoogle Scholar
  16. Jayaraman A, Örnek D, Duarte DA, Lee C-C, Mansfeld FB, Wood TK (1999c) Axenic aerobic biofilms inhibit corrosion of copper and aluminum. Appl Microbiol Biotechnol 52:787–790PubMedGoogle Scholar
  17. Licina GJ (1988) Sourcebook for microbiologically influenced corrosion in nuclear power plants RP2812-2. Electric Power Research Institute, Palo Alto, Calif.Google Scholar
  18. Little B, Ray R (2002) A perspective on corrosion inhibition by biofilms. Corrosion 58:424–428Google Scholar
  19. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.Google Scholar
  20. Mansfeld F (1976) The polarization resistance technique for measuring corrosion currents. In: Fontana MG, Staehle RW(eds) Advances in corrosion science and technology. Plenum Press, New York, 163–262Google Scholar
  21. Mansfeld F (1995) Use of electrochemical impedance spectroscopy for the study of corrosion protection by polymer coatings. J Appl Electrochem 25:187–202Google Scholar
  22. Mansfeld F, Tsai CH, Shih H (1992) Software for simulation and analysis of electrochemical impedance spectroscopy (EIS) data. ASTM Spec Tech Publ 1154:186–196Google Scholar
  23. Miller JDA (1981) Metals. In: Rose AH (ed) Microbial biodeterioration. Academic Press, New York, 149–202Google Scholar
  24. Nagarajan V, Albertson H, Chen M, Ribbe J (1992) Modular expression and secretion vectors for Bacillus subtilis. Gene 114:121–126PubMedGoogle Scholar
  25. Odom JM (1990) Industrial and environmental concerns with sulfate-reducing bacteria. ASM News 56:473–476Google Scholar
  26. Paddon CJ, Vasantha N, Hartley RW (1989) Translation and processing of Bacillus amyloliquefaciens extracellular RNase. J Bacteriol 171:1185–1187PubMedGoogle Scholar
  27. Pankhurst ES (1968) Significance of sulphate-reducing bacteria to the gas industry: a review. J Appl Bacteriol 31:179–193Google Scholar
  28. Potekhina JS, Sherisheva NG, Povetkina LP, Pospelov AP, Rakitina TA, Warnecke F, Gottschalk G (1999) Role of microorganisms in corrosion inhibition of metals in aquatic habitats. Appl Microbiol Biotechnol 52:639–646CrossRefGoogle Scholar
  29. Romeo D, Skerlavaj B, Bolognesi M, Gennaro R (1988) Structure and bactericidal activity of an antibiotic dodecapeptide purified from bovine neutrophils. J Biol Chem 263:9573–9575PubMedGoogle Scholar
  30. Wu X-C, Lee W, Tran L, Wong S-L (1991) Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. J Bacteriol 173:4952–4958PubMedGoogle Scholar
  31. Zhang L, Rozek A, Hancock REW (2001) Interaction of cationic antimicrobial peptides with model membranes. J Biol Chem 276:35714–35722CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • R. Zuo
    • 1
  • D. Örnek
    • 1
    • 5
  • B. C. Syrett
    • 2
  • R. M. Green
    • 3
  • C.-H. Hsu
    • 4
  • F. B. Mansfeld
    • 4
  • T. K. Wood
    • 1
  1. 1.Departments of Chemical Engineering & Molecular and Cell BiologyUniversity of ConnecticutStorrsUSA
  2. 2.Electric Power Research InstitutePalo AltoUSA
  3. 3.TMI Nuclear Generation StationMiddletownUSA
  4. 4.Department of Materials Science & EngineeringUniversity of Southern CaliforniaLos AngelesUSA
  5. 5.Genzyme CorporationCambridgeUSA

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