Antonie van Leeuwenhoek

, Volume 74, Issue 1–3, pp 59–70 | Cite as

Surface-active lipids in rhodococci

  • Siegmund Lang
  • Jim C PhilpEmail author


Like other hydrocarbon-oxidising bacteria, rhodococci respond to the presence of alkanes by producing biosurfactant molecules to improve their ability to utilise these hydrophobic compounds as growth substrates. In the rhodococci these surfactants are predominantly glycolipids, the majority of which remain cell-bound during unrestricted growth. Most work has been done on the trehalose mycolates formed by Rhodococcus erythropolis, but nitrogen- limited conditions lead to the production of anionic trehalose tetraesters also.

As surfactants, these compounds, whether purified or in crude form, are able to reduce the surface tension of water from 72 mN m-1 to a low of 26, thus making them among the most potent biosurfactants known. They are also able to reduce the interfacial tension between water and a hydrophobic phase (e.g. n- hexadecane) from 43 mN m-1 to values less than one (Table 1). Biosurfactants have about a ten- to 40-fold lower critical micelle concentration than synthetic surfactants. Such properties suggest a range of industrial applications, where a variety of surface-active characteristics are appropriate. Interest in biosurfactants as industrial chemicals results from the toxicity of many petrochemical-derived surfactants. Currently world-wide surfactant production is on a very large scale, and the demand for them is increasing. However, the drive towards less environmentally damaging chemicals makes biosurfactants attractive as they have lower toxicity.

The reason they have not achieved a significant market share is the cost of production, which is considerably higher than for synthetic surfactants. This problem is being addressed using several strategies. An approach where there is great scope for improvement with the rhodococci is an understanding of the genetic basis of glycolipid production, which is largely unknown. They may find applications in the near future in the environmental remediation industries, where the requirement for purified molecules is of less importance.

This review summarises knowledge of the chemistry, biochemistry and production of Rhodococcus surface-active lipids. Where they have been used, or there is potential for use, in industrial applications is discussed.

biosurfactants glycolipids remediation rhodococci 


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  1. Asmer H-J (1991) Trehalose lipid formation: studies on substrate specificity and biochemical acylation of trehalose using the marine bacterium Arthrobacter sp. EK1. PhD thesis, Technical University of Braunschweig, GermanyGoogle Scholar
  2. Asselineau C & Asselineau J (1978) Trehalose-containing glycolipids. Prog. Chem. Fats other Lipids 16: 59–99Google Scholar
  3. Bai G, Brusseau ML & Miller RM (1997) Influence of rhamnolipid biosurfactant on the transport of bacteria through a sandy soil. Appl. Environ. Microbiol. 63: 1866–1873Google Scholar
  4. Banat IM, Samarah N, Murad M, Horne R & Banerjee S (1991) Biosurfactant production and use in oil tank clean-up. World J. Microbiol. Biotechnol. 7: 80–88Google Scholar
  5. Batrakov SG, Rozynov BV, Koronelli TV & Bergelson LD (1981) Two novel types of trehalose lipids. Chem. Phys. Lipids 29: 241–266Google Scholar
  6. Bouchez M, Blanchet D & Vandecasteele, J-P (1995) Substrate availability in phenanthrene biodegradation: transfer mechanism and influence on metabolism. Appl. Microbiol. Biotechnol. 43: 952–960Google Scholar
  7. Brennan PJ, Lehane DP & Thomas DW (1970) Acylglucoses of the corynebacteria and mycobacteria. Eur. J. Biochem. 13: 117–123Google Scholar
  8. Chiou CT, Porter PE, & Schmeddling DW (1983) Partition equilibria of nonionic organic compounds between soil and organic matter and water. Environ. Sci. Technol. 17: 227–231Google Scholar
  9. Clint JH (1992) Surfactant Aggregation. Blackie & Son Ltd, GlasgowGoogle Scholar
  10. Desai JD & Banat IM (1997) Microbial production of surfactants and their commercial potential. Microbiol. Mol. Biol. Rev. 61: 47–64Google Scholar
  11. Espuny MJ, Egido S, Mercad & #x00E9; ME & Manresa A (1995) Characterization of trehalose tetraester produced by a waste lubricant oil degrader Rhodococcus sp. Toxicol. Environ. Chem. 48: 83–88Google Scholar
  12. Espuny MJ, Egido S, Rodón I, Manresa A & Mercad & #x00E9; ME (1996) Nutritional requirements of a biosurfactant producing strain Rhodococcus sp. 51T7. Biotechnol. Lett. 18: 521–526Google Scholar
  13. Fiechter A (1992) Biosurfactants: moving towards industrial application. Tibtech. 10: 208–217Google Scholar
  14. Finnerty WR & Singer MEV (1984) A microbial biosurfactant – physiology, biochemistry and applications. Dev. Ind. Microbiol. 25: 31–40Google Scholar
  15. Fougias E & Forster CF (1994) Rhodococcus rubra in relation to stable foams in activated sludge. Process Biochem. 29: 553–557Google Scholar
  16. Goodfellow M, Davenport R, Stainsby FM & Curtis TP (1997) Actinomycete diversity associated with foaming in activated sludge plants. J. Ind. Microbiol. 17: 268–280Google Scholar
  17. Guerra-Santos L, K & #x00E4;ppeli O & Fiechter A (1984) Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source. Appl. Environ. Microbiol. 48: 301–305Google Scholar
  18. Haferburg D, Hommel RK, Claus R & Kleber H-P (1986) Extracellular microbial lipids as biosurfactants. Adv. Biochem. Eng. / Biotechnol. 33: 53–93Google Scholar
  19. Herman DC, Artiola JF & Miller RM (1995) Removal of cadmium, lead and zinc from soil by a rhamnolipid biosurfactant. Environ. Sci. Technol. 29: 2280–2285Google Scholar
  20. Hommel RK (1990) Formation and physiological role of biosurfactants produced by hydrocarbon-utilising microorganisms. Biodegradation 1: 107–119Google Scholar
  21. Hommel RK & Ratledge C (1993) Biosynthetic mechanisms of low molecular weight surfactants and their precursor molecules. In: Kosaric N (Ed) Biosurfactants: Production, Properties, Applications, Surfactant Science Series, Vol 48 (pp 3–63). Marcel Dekker Inc, New YorkGoogle Scholar
  22. Ishigami Y, Suzuki S, Funada T, Chino M, Uchida Y & Tabuchi T (1987) Surface-active properties of succinoyl trehalose lipids as microbial biosurfactants. J. JPN. Oil Chem. Soc. (YUKAGAKU) 36: 847–851Google Scholar
  23. Isoda H, Kitamoto D, Shinmoto H, Matsumara M & Nakahara T (1997) Microbial extracellular glycolipid induction of differentiation and inhibition of the protein kinase C activity of human promyelocytic leukemia cell line HL60. Biosci. Biotechnol. Biochem. 61: 609–614Google Scholar
  24. Ivshina IB, Kuyukina MS, Philp JC & Christofi N (1998) Oil desorption from mineral and organic materials using biosurfactant complexes produced by Rhodococcus species. World J. Microbiol. Biotechnol. (in press)Google Scholar
  25. Jain DK, Lee H & Trevors JT (1992) Effect of addition of Pseudomonas aeruginosa UG2 inocula or biosurfactants on biodegradation of selected hydrocarbons in soil. J. Ind. Microbiol. 10: 87–93Google Scholar
  26. Khan AR & Forster CF (1990) An investigation into the stability of foams related to the activated sludge process. Enz. Microb. Technol. 12: 788–793Google Scholar
  27. Kim, J-S, Powalla M, Lang S, Wagner F, L & #x00FC;nsdorf H & Wray V (1990) Microbial glycolipid production under nitrogen limitation and resting cell conditions. J. Biotechnol. 13: 257–266Google Scholar
  28. Kretschmer A & Wagner F (1983) Characterization of biosynthetic intermediates of trehalose dicorynomycolates from Rhodococcus erythropolis grown on n-alkanes. Biochim. Biophys. Acta 753: 306–313Google Scholar
  29. Kretschmer A, Bock H & Wagner F (1982) Chemical and physical characterization of interfacial-active lipids from Rhodococcus erythropolis grown on n-alkanes. Appl. Environ. Microbiol. 44: 864–870Google Scholar
  30. Kurane R & Tomizuka N (1992) Towards new biomaterial produced by microorganism — bioflocculant and bioabsorbent. Nippon Kagaku Kaishi 5: 453–463Google Scholar
  31. Kurane R, Hatamochi K, Kakuno T, Kiyohara M, Tajima T, Hirano M & Taniguchi Y (1995) Chemical structure of lipid bioflocculant produced by Rhodococcus erythropolis. Biosci. Biotechnol. Biochem. 59: 1652–1656Google Scholar
  32. Laha S & Luthy RG (1992) Effects of nonionic surfactants on the solubilisation and mineralisation of phenanthrene in soil-water systems. Biotechnol. Bioeng. 40: 1367–1380Google Scholar
  33. Lang S & Wagner F (1993) Biological activities of biosurfactants. In: Kosaric N (Ed) Biosurfactants — Production, Properties and Applications. Surfactant Science Series, Vol, 48. Marcel Dekker Inc, New YorkGoogle Scholar
  34. Lang S, Brakemeier A, Schlotterbeck A & Wagner F (1996) Biotechnological synthesis of surface-active glycolipids. Eurocosmetics 4: 41–45Google Scholar
  35. Li Z-Y, Lang S, Wagner F, Witte L & Wray V (1984) Formation and identification of interfacial-active glycolipids from resting microbial cells. Appl. Environ. Microbiol. 48: 610–617Google Scholar
  36. Liu Z, Jacobson, AM & Luthy RG (1995) Biodegradation of naphthalene in aqueous nonionic surfactant systems. Appl. Environ. Microbiol. 61: 145–151Google Scholar
  37. McNeil MM & Brown JM (1994) The medically important aerobic actinomycetes: epidemiology and microbiology. Clin.Microbiol. Rev. 7: 357–417Google Scholar
  38. Mercad & #x00E9; ME, Monleón L, de Andr & #x00E9;s C, Rodón I, Martinez E, Espuny MJ & Manresa A (1996) Screening and selection of surfactant-producing bacteria from waste lubricating oil. J. Appl. Bacteriol. 81: 161–166Google Scholar
  39. Miller RM (1995) Biosurfactant-facilitated remediation of metal-contaminated soils. Environ. Health Perspect. 103: 59–62Google Scholar
  40. Mori T, Sakai Y, Honda K, Yano I & Hashimoto S (1988) Stable abnormal foam in activated sludge process produced by Rhodococcus sp. with strong hydrophobic property. Environ. Technol. Lett. 9: 1041–1048Google Scholar
  41. Neu TR (1996) Significance of bacterial surface-active compounds in interaction of bacteria with interfaces. Microbiol. Rev. 60: 151–166Google Scholar
  42. Oberbremer A & M & #x00FC;ller-Hurtig R (1989) Aerobic, step-wise hydrocarbon degradation and formation of biosurfactants by an original soil population in a stirred reactor. Appl. Microbiol. Biotechnol. 31: 582–586Google Scholar
  43. Oberbremer A, M & #x00FC;ller-Hurtig R & Wagner F (1990) Effect of the addition of microbial surfactants on hydrocarbon degradation in a soil population in a stirred reactor. Appl. Microbiol. Biotechnol. 32: 485–489Google Scholar
  44. Passeri A, Lang S & Wagner F (1991) Marine biosurfactants. II. Production and characterisation of an anionic trehalose tetraester from the marine bacterium Arthrobacter sp. EK1. Z.Naturforsch. 46c: 204–209Google Scholar
  45. Poremba K, Gunkel W, Lang S & Wagner F (1991) Marine biosurfactants. III. Toxicity testing with marine microorganisms and comparison with synthetic surfactants. Zeit. Naturforsch. 46c: 210–216Google Scholar
  46. Ramsay BA, Cooper DG, Margaritis A & Zajic JE (1983) Rhodochrous bacteria: biosurfactant production and demulsifying ability. In: Zajic JE, Cooper DG, Jack TR, Kosaric N (Eds.) Microbial Enhanced Oil Recovery (pp 61–65). PennWell Books, Tulsa, OklahomaGoogle Scholar
  47. Rapp P, Bock H, Wray V & Wagner F (1979) Formation, isolation and characterisation of trehalose dimycolates from Rhodococcus erythropolis grown on n-alkanes. J. Gen. Microbiol. 115: 491–503Google Scholar
  48. Ristau E & Wagner F (1983) Formation of novel anionic trehalose tetraesters from Rhodococcus erythropolis under growth-limiting conditions. Biotechnol. Lett. 5: 95–100Google Scholar
  49. Rosenberg E (1993) Exploiting microbial growth on hydrocarbons —new markets. Tibtech. 11: 419–423Google Scholar
  50. Scheibenbogen K, Zytner RG, Lee H & Trevors JT (1994) Enhanced removal of selected hydrocarbons from soil by Pseudomonas aeruginosa UG2 biosurfactants and some chemical surfactants. J. Chem. Technol. Biotechnol. 59: 53–59Google Scholar
  51. Singer MEV & Finnerty WR (1990) Physiology of biosurfactant synthesis by Rhodococcus species H13-A. Can. J. Microbiol. 36: 741–745Google Scholar
  52. Singer MEV, Finnerty WR & Tunelid A (1990) Physical and chemical properties of a biosurfactant synthesized by Rhodococcus species H13-A. Can. J. Microbiol. 36: 746–750Google Scholar
  53. Stoecker MA, Machlin SM & Staley JT (1996) Cloning and characterisation of genes required for the production of an emulsion-stabilizing capsule in Rhodococcus erythropolis strain NO1–1. Abstract presented at the 96th General Meeting of the American Society of MicrobiologyGoogle Scholar
  54. Sunairi M, Iwabuchi Y, Yoshizawa Y, Murooka H, Morisaki H & Nakajima M (1997) Cell surface hydrophobicity and scum formation of Rhodococcus rhodochrous strains with different colony morphologies. J. Appl. Microbiol. 82: 204–210Google Scholar
  55. Suzuki T, Tanaka H & Itoh S (1974) Sucrose lipids of Arthrobacteria, Corynebacteria and Nocardia grown on sucrose. Agric. Biol. Chem. 38: 557–563Google Scholar
  56. Tan H, Champion JT, Artiola JF, Brusseau ML & Miller RM (1994) Complexation of cadmium by a rhamnolipid biosurfactant. Environ. Sci. Technol. 28: 2402–2406Google Scholar
  57. Tiehm A (1994) Degradation of polycyclic aromatic aromatic hydrocarbons in the presence of synthetic surfactants. Appl. Environ. Microbiol. 60: 258–263Google Scholar
  58. Uchida Y, Misawa S, Nakahara T & Tabuchi T (1989a) Factors affecting the production of succinoyl trehalose lipids by Rhodococcus erythropolis SD-74 grown on n-alkanes. Agric. Biol. Chem. 53: 765–769Google Scholar
  59. Uchida Y, Tsuchiya R, Chino M, Hirano J & Tabuchi T (1989b) Extracellular accumulation of mono-and di-succinoyl trehalose lipids by a strain of Rhodococcus erythropolis grown on nalkanes. Agric. Biol. Chem. 53: 757–763Google Scholar
  60. van Dyke MI, Gulley SL, Lee H & Trevors JT (1993) Evaluation of microbial surfactants for recovery of hydrophobic pollutants from soil. J. Ind. Microbiol. 11: 163–170Google Scholar
  61. Volkering F, Breure AM, Sterkenburg A & van Andel JG (1992) Microbial degradation of polycyclic aromatic hydrocarbons: effect of substrate availability on bacterial growth kinetics. Appl. Microbiol. Biotechnol. 36: 548–552Google Scholar
  62. Wagner F, Behrendt U, Bock H, Kretschmer A, Lang S & Syldatk C (1983) Production and chemical characterisation of surfactants from Rhodococcus erythropolis and Pseudomonas sp. MUB grown on hydrocarbons. In: Zajic JE, Cooper DG, Jack TR & Kosaric N (Eds) Microbial Enhanced Oil Recovery (pp 55–60). PennWell Publishing Company, TulsaGoogle Scholar
  63. Wagner F, Kim J-S, Lang S, Li Z-Y, Marwede G, Matulovic U, Ristau E & Syldatk C (1984) Production of surface active anionic glycolipids by resting and immobilized microbial cells. In: Proceedings of 3rd European Congress on Biotechnology, Vol I (pp 3–8), Verlag Chemie, Weinheim, GermanyGoogle Scholar
  64. Wodzinski RS & Bertolini D (1972) Physical state in which naphthalene and bibenzyl are utilised by bacteria. Appl. Microbiol. 23: 1077–1081Google Scholar
  65. Wodzinski RS & Coyle JE (1974) Physical state of phenanthrene for utilisation by bacteria. Appl. Microbiol. 27: 1081–1084Google Scholar
  66. Zhang Y & Miller RM (1992) Enhanced octadecane dispersion and biodegradation by a Pseudomonas rhamnolipid surfactant (biosurfactant). Appl. Environ. Microbiol. 58: 3276–3282Google Scholar
  67. Zhang Y & Miller RM (1995) Effect of rhamnolipid (biosurfactant) structure on solubilization and biodegradation of n-alkanes. Appl. Environ. Microbiol. 61: 2247–2251Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  1. 1.Department of Biochemistry and Biotechnology, Biotechnology GroupTechnical University BraunschweigBRAUNSCHWEIGGermany
  2. 2.Department of Biological SciencesNapier UniversityScotlandU. K.

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