, Volume 17, Issue 4, pp 669–675 | Cite as

Evidence for surfactant production by the haloarchaeon Haloferax sp. MSNC14 in hydrocarbon-containing media

  • Ikram Djeridi
  • Cécile Militon
  • Vincent Grossi
  • Philippe CunyEmail author
Original Paper


The potential for surfactant production by the extreme halophilic archaeon Haloferax sp. MSNC14 in the presence of individual hydrocarbon substrates was studied. This strain was selected for its ability to grow on different types of hydrocarbons at high NaCl concentrations. Linear (n-heptadecane or C17) and isoprenoid (pristane) alkanes, a polyaromatic hydrocarbon (phenanthrene) and ammonium acetate (highly water-soluble control compound) were used as growth substrates. The adherence potential was demonstrated by the ability of the cells to adhere to liquid or solid hydrocarbons. The biosurfactant production was indicated by the reduction of the surface tension (ST) and by the emulsification activity (EA) of cell-free supernatants. Growth on acetate was accompanied by a low EA (lower than 0.1) and a high ST (~70 mN/m), whereas an important EA (up to 0.68 ± 0.08) and a reduction of ST (down to 32 ± 2.3 mN/m) were observed during growth on the different hydrocarbons. Both ST and EA varied with the growth phase. The adhesion to hydrocarbons was higher when cells were grown on C17 (by 60–70 %) and pristane (by 30–50 %) than on phenanthrene (~25 %). The results demonstrated that strain MNSC14 was able to increase the bioavailability of insoluble hydrocarbons, thus facilitating their uptake and their biodegradation even at high salt concentration.


Haloferax volcanii MNSC14 Hydrocarbon biodegradation Emulsifying activity Surface tension Adherence 



This work was funded by Grants from the French National Program EC2CO “BIOHYDEX (BIOdégradation des HYDrocarbures dans les milieux EXtrêmes’’). We thank the anonymous reviewers for helpful comments and suggestions.


  1. Abbasnezhad H, Foght JM, Gray MR (2011) Adhesion to the hydrocarbon phase increases phenanthrene degradation by Pseudomonas fluorescens LP6a. Biodegradation 22:485–496PubMedCrossRefGoogle Scholar
  2. Al-Mailem DM, Sorkhoh NA, Al-Awadhi H, Eliyas M, Radwan SS (2010) Biodegradation of crude oil and pure hydrocarbons by extreme halophilic archaea from hypersaline coasts of the Arabian Gulf. Extremophiles 14:321–328PubMedCrossRefGoogle Scholar
  3. Al-Tahhan RA, Sandrin TR, Bodour AA, Maier RM (2000) Rhamnolipid-induced removal of lipopolysaccharide from Pseudomonas aeruginosa: effect on cell surface properties and interaction with hydrophobic substrates. Appl Environ Microbiol 66:3262–3268PubMedCrossRefGoogle Scholar
  4. Andrei AS, Banciu HL, Oren A (2012) Living with salt: metabolic and phylogenetic diversity of archaea inhabiting saline ecosystems. FEMS Microbiol Lett 330:1–9PubMedCrossRefGoogle Scholar
  5. Anton J, Meseguer I, Rodríguez-Valera F (1988) Production of an Extracellular Polysaccharide by Haloferax mediterranei. Appl Environ Microbiol 54:2381–2386PubMedGoogle Scholar
  6. Bertrand JC, Almallah M, Acquaviva M, Mille G (1990) Biodegradation of hydrocarbons by an extremely halophilic archaebacterium. Lett Appl Microbiol 11:260–263CrossRefGoogle Scholar
  7. Bonfá MRL, Grossman MJ, Mellado E, Durrant LR (2011) Biodegradation of aromatic hydrocarbons by Haloarchaea and their use for the reduction of the chemical oxygen demand of hypersaline petroleum produced water. Chemosphere 84:1671–1676PubMedCrossRefGoogle Scholar
  8. Bury SJ, Miller CA (1993) Effect of micellar solubilization on biodegradation rates of hydrocarbons. Environ Sci Technol 27:104–110CrossRefGoogle Scholar
  9. Christensen BE (1989) The role of extracellular polysaccharides in biofilms. J Biotechnol 10:181–202CrossRefGoogle Scholar
  10. Cuny P, Gilewicz M, Faucet J, Acquaviva M, Bertrand JC (1999) Enhanced Biodegradation of phenanthrene by a marine bacterium in presence of a synthetic surfactant. Lett Appl Microbiol 29:242–245PubMedCrossRefGoogle Scholar
  11. Cuny P, Acquaviva M, Gilewicz M (2004) Phenanthrene degradation, emulsification and surface tension activities of a Pseudomonas putida strain isolated from a coastal oil contaminated microbial mat. Ophelia 58:283–287CrossRefGoogle Scholar
  12. Dalvi S, Azetsu S, Patrauchan MA, Akta DF, Fathepure BZ (2012) Proteogenomic elucidation of the initial steps in the benzene degradation pathway of a novel halophile, Arhodomonas sp. strain Rozel, isolated from a hypersaline environment. Appl Environ Microbiol 78:7309–7316PubMedCrossRefGoogle Scholar
  13. Das K, Mukherjee AK (2007) Differential utilization of pyrene as the sole source of carbon by Bacillus subtilis and Pseudomonas aeruginosa strains: role of biosurfactants in enhancing bioavailability. J Appl Microbiol 102:195–203PubMedCrossRefGoogle Scholar
  14. Dastgheib SMM, Amoozegar MA, Khajeh K, Ventosa A (2011) A halotolerant Alcanivorax sp. strain with potential application in saline soil remediation. Appl Microbiol Biotechnol 90:305–312PubMedCrossRefGoogle Scholar
  15. De Medeiros Rocha R, Costa DFS, Lucena-Filho MA, Bezerra RM, Medeiros DHM, Azevedo-Silva AM, Araújo CN, Xavier-Filho L (2012) Brazilian solar saltworks—ancient uses and future possibilities. Aquat Biosyst 8:8PubMedCrossRefGoogle Scholar
  16. Dewulf J, Drijvers D, Langenhove HV (1995) Measurement of Henry’s law constant as function of temperature and salinity for the low temperature range. Atmos Environ 29:323–333CrossRefGoogle Scholar
  17. Emerson D, Chauhan S, Oriel P, John A (1994) Breznak Haloferax sp. D1227, a halophilic Archaeon capable of growth on aromatic compounds. Arch Microbiol 161:445–452CrossRefGoogle Scholar
  18. Erdoğmuş SF, Mutlu B, Korcan SE, Güven K, Konuk M (2013) Aromatic hydrocarbon degradation by halophilic archaea isolated from Çamaltı Saltern, Turkey. Water Air Soil Pollut 224:1449Google Scholar
  19. Górna H, Lawniczak L, Zgola-Grzeskowiak A, Kaczorek E (2011) Differences and dynamic changes in the cell surface properties of three Pseudomonas aeruginosa strains isolated from petroleum-polluted soil as a response to various carbon sources and the external addition of rhamnolipids. Bioresour Technol 102:3028–3033PubMedCrossRefGoogle Scholar
  20. Goswami PC, Singh HD, Bhagat SD, Baruah JN (1983) Mode of uptake of insoluble solid substrates by microrganisms.1. Sterol uptake by an arthtobacter species. Biotechnol Bioeng 25:2929–2943PubMedCrossRefGoogle Scholar
  21. Haritash AK, Kaushik CP (2009) Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): a review. J Hazard Mater 169:1–15PubMedCrossRefGoogle Scholar
  22. He H, Yang Y, Xia JL, Ding JN, Zhao XJ, Nie ZY (2008) Growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis YN-25 grown on different substrates. Trans Nonferrous Metals Soc China 18:1374–1378CrossRefGoogle Scholar
  23. Hommel RK (1990) Formation and physiological role of biosurfactants produced by hydrocarbon-utilizing microorganisms. Biosurfactants in hydrocarbon utilization. Biodegradation 1:107–119PubMedCrossRefGoogle Scholar
  24. Hwang G, Ban YM, Lee CH, Chung CH, Ahn IS (2008) Adhesion of Pseudomonas putida NCIB 9816–4 to a naphthalene-contaminated soil. Colloids Surf B 62:91–96CrossRefGoogle Scholar
  25. Kebbouche-Gana S, Gana ML, Khemili S, Fazouane-Naimi F, Bouanane NA, Penninckx M, Hacene H (2009) Isolation and characterization of halophilic Archaea able to produce biosurfactants. J Ind Microbiol Biotechnol 36:727–738PubMedCrossRefGoogle Scholar
  26. Kulichevskaya IS, Milekhina EI, Borzenkov IA, Zvyagintseva IS, Belyaev SS (1991) Oxidation of petroleum-hydrocarbons by extremely halophilic archaebacteria. Microbiology 60:596–601Google Scholar
  27. Kumar M, Vladimir L, Materano ADS, Ilzins OA, Luis L (2008) Biosurfactant production and hydrocarbon-degradation by halotolerant and thermotolerant Pseudomonas sp. World J Microbiol Biotechnol 24:1047–1057CrossRefGoogle Scholar
  28. Lefebvre O, Moletta R (2006) Treatment of organic pollution in industrial saline wastewater: a literature review. Water Res 40:3671–3682PubMedCrossRefGoogle Scholar
  29. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  30. Nicolaus B, Lama L, Esposito E, Manca MC, Improta R, Bellitti MR, Duckworth AW, Grant WD, Gambacorta A (1999) Haloarcula spp able to biosynthesize exo-endopolymers. J Ind Microbiol Biotechnol 23:489–496CrossRefGoogle Scholar
  31. Nicolaus B, Moriello VS, Lama L, Poli A, Gambacorta A (2004) Polysaccharides from extremophilic microorganisms. Orig Life Evol Biospheres 34:159–169CrossRefGoogle Scholar
  32. Nomura T, Yoshihara A, Nagao T, Tokumoto H, Konishi Y (2007) Effect of the surface characteristics of Methanosarcina barkeri on immobilization to support materials. Adv Power Technol 18:489–501CrossRefGoogle Scholar
  33. Oren A (2001) The order Halobacteriales, p. 2, 6–19. In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E (eds) The prokaryotes. A handbook on the biology of bacteria: ecophysiology, isolation, identification, applications, 3rd edn. Springer, New York (Online)Google Scholar
  34. Oren A (2002a) Diversity of halophilic microorganisms: environments, phylogeny, physiology, and applications. J Ind Microbiol Biotechnol 28:56–63PubMedGoogle Scholar
  35. Oren A (2002b) Halophilic microorganisms and their environments. Kluwer, Dordrecht, p 575CrossRefGoogle Scholar
  36. Oren A (2002c) Molecular ecology of extremely halophilic Archaea and Bacteria. FEMS Microbiol Ecol 39:1–7PubMedCrossRefGoogle Scholar
  37. Robinson JL, Pyzyna B, Atrasz RG, Henderson CA, Morrill KL, Burd AM, DeSoucy E, Fogleman RE III, Naylor JB, Steele SM, Elliott DR, Leyva KJ, Shand RF (2005) Growth kinetics of extremely halophilic Archaea (Family Halobacteriaceae) as revealed by arrhenius plots. J Bacteriol 187:923–929PubMedCrossRefGoogle Scholar
  38. Romero-Viana L, Kienel U, Sachse D (2012) Lipid biomarker signatures in a hypersaline lake on Isabel Island (Eastern Pacific) as a proxy for past rainfall anomaly (1942–2006 AD). Palaeogeogr Palaeoclimatol Palaeoecol 350–352:49–61CrossRefGoogle Scholar
  39. Rosenberg M (2006) Microbial adhesion to hydrocarbons: twenty-five years of doing MATH. FEMS Microbiol Lett 262:129–134PubMedCrossRefGoogle Scholar
  40. Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons—a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33CrossRefGoogle Scholar
  41. Roy PK, Singh HD, Bhagat SD, Baruah JN (1979) Characterisation of hydrocarbon emulsification and solubilisation occurring during the growth of Endomycopsis lipolytica on hydrocarbons. Biotechnol Bioeng 21:955–974CrossRefGoogle Scholar
  42. Sei A, Fathepure BZ (2009) Biodegradation of BTEX at high salinity by an enrichment culture from hypersaline sediments of Rozel Point at Great Salt Lake. J Appl Microbiol 107:2001–2008PubMedCrossRefGoogle Scholar
  43. Tapilatu YH, Grossi V, Acquaviva M, Militon C, Bertrand JC, Cuny P (2010) Isolation of hydrocarbon-degrading extremely halophilic archaea from an uncontaminated hypersaline pond (Camargue, France). Extremophiles 14:225–231PubMedCrossRefGoogle Scholar
  44. Ward OP (2010) Microbial biosurfactants and biodegradation. In: Sen R (ed) Biosurfactants. Advances in experimental medicine and biology, vol 672, pp 65–74Google Scholar
  45. Zhang Y, Miller RM (1995) Effect of rhamnolipid (biosurfactant) structure on solubilization and biodegradation of n-alkanes. Appl Environ Microbiol 61:2247–2251PubMedGoogle Scholar

Copyright information

© Springer Japan 2013

Authors and Affiliations

  • Ikram Djeridi
    • 1
  • Cécile Militon
    • 1
  • Vincent Grossi
    • 2
  • Philippe Cuny
    • 1
    Email author
  1. 1.Aix-Marseille Université, Université du Sud Toulon-Var, CNRS/INSU, IRD, MIO, UM 110Marseille Cedex 09France
  2. 2.CNRS, UMR5276, Laboratoire de Géologie de LyonUniversité Lyon 1VilleurbanneFrance

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