Applied Microbiology and Biotechnology

, Volume 75, Issue 1, pp 175–186 | Cite as

Influence of growth medium on cometabolic degradation of polycyclic aromatic hydrocarbons by Sphingomonas sp. strain PheB4

Environmental Biotechnology


The influence of growth medium on cometabolic degradation of polycyclic aromatic hydrocarbons (PAHs) was investigated when Sphingomonas sp. strain PheB4 isolated from surface mangrove sediments was grown in either phenanthrene-containing mineral salts medium (PMSM) or nutrient broth (NB). The NB-grown culture exhibited a more rapid cometabolic degradation of single and mixed non-growth substrate PAHs compared to the PMSM-grown culture. The concentrations of PAH metabolites were also lower in NB-grown culture than in PMSM-grown culture, suggesting that NB-grown culture removed metabolites at a faster rate, particularly, for metabolites produced from cometabolic degradation of a binary mixture of PAHs. Cometabolic pathways of single PAH (anthracene, fluorene, or fluoranthene) in NB-grown culture showed similarity to that in PMSM-grown culture. However, cometabolic pathways of mixed PAHs were more diverse in NB-grown culture than that in PMSM-grown culture. These results indicated that nutrient rich medium was effective in enhancing cometabolic degradation of mixed PAHs concomitant with a rapid removal of metabolites, which could be useful for the bioremediation of mixed PAHs contaminated sites using Sphingomonas sp. strain PheB4.


Biodegradation Cometabolism Solid phase microextraction Mangrove ecosystems 


  1. Bezalel L, Hadar Y, Fu P, Freeman JP, Cerniglia CE (1996) Initial oxidation products in the metabolism of pyrene, anthracene, fluorene, and dibenzothiophene by the white rot fungus Pleurotus ostreatus. Appl Environ Microbiol 62:2554–2559Google Scholar
  2. Bouchez M, Blanchet D, Vandecasteele J-P (1995) Degradation of polycyclic aromatic hydrocarbons by pure strains and by defined strain associations: inhibition phenomena and cometabolism. Appl Microbiol Biotechnol 43:156–164CrossRefGoogle Scholar
  3. Breedveld GD, Sparrevik M (2000) Nutrient-limited biodegradation of PAH in various soil strata at a creosote contaminated site. Biodegradation 11:391–399CrossRefGoogle Scholar
  4. Carmichael LM, Pfaender FK (1997) The effect of inorganic and organic supplements on the microbial degradation of phenanthrene and pyrene in soils. Biodegradation 8:1–13CrossRefGoogle Scholar
  5. Casellas M, Grifoll M, Bayona JM, Solanas AM (1997) New metabolites in the degradation of fluorene by Arthrobacter sp. strain F101. Appl Environ Microbiol 63:819–826Google Scholar
  6. Cerniglia CE (1993) Biodegradation of polycyclic aromatic hydrocarbons. Curr Opin Biotechnol 4:331–338CrossRefGoogle Scholar
  7. Cheung P-Y, Kinkle BK (2005) Effects of nutrients and surfactants on pyrene mineralization and Mycobacterium spp. populations in contaminated soil. Soil Biol Biochem 37:1401–1405CrossRefGoogle Scholar
  8. Criddle CS (1993) The kinetics of cometabolism. Biotechnol Bioeng 41:1048–1056CrossRefGoogle Scholar
  9. Dean-Ross D, Moody J, Cerniglia CE (2002) Utilization of mixtures of polycyclic aromatic hydrocarbons by bacteria isolated from contaminated sediment. FEMS Microbiol Ecol 41:1–7CrossRefGoogle Scholar
  10. Demanèche S, Meyer C, Micoud J, Louwagie M, Willison JC, Jouanneau Y (2004) Identification and functional analysis of two aromatic-ring-hydroxylating dioxygenase from a Sphingomonas strain that degrade various polycyclic aromatic hydrocarbons. Appl Environ Microbiol 70:6714–6725CrossRefGoogle Scholar
  11. Guha S, Peters CA, Jaffe PR (1999) Multisubstrate biodegradation kinetics of naphthalene, phenanthrene and pyrene mixtures. Biotechnol Bioeng 65:491–499CrossRefGoogle Scholar
  12. Ho Y, Jackson M, Yang Y, Mueller JG, Pritchard PH (2000) Characterization of fluoranthene- and pyrene-degrading bacteria isolated from PAH-contaminated soils and sediments. J Ind Microbiol Biotechnol 24:100–112CrossRefGoogle Scholar
  13. Horvath RS (1972) Microbial co-metabolism and the degradation of organic compounds in nature. Bacteriol Rev 36:146–155Google Scholar
  14. Kazunga C, Aitken MD, Gold A, Sangaiah R (2001) Fluoranthene-2,3- and -1,5-diones are novel products from the bacterial transformation of fluoranthene. Environ Sci Technol 35:917–922CrossRefGoogle Scholar
  15. Kelley I, Freeman JP, Evans E, Cerniglia CE (1993) Identification of metabolites from the degradation of fluoranthene by Mycobacterium sp. strain PYR-1. Appl Environ Microbiol 59:800–806Google Scholar
  16. Kim E, Zylstra GJ, Freeman JP, Heinze TM, Deck J, Cerniglia CE (1997) Evidence for the role of 2-2-hydroxychromene-2-carboxylate isomerase in the degradation of anthracene by Sphingomonas yanoikuyae B1. FEMS Microbiol Lett 153:479–484CrossRefGoogle Scholar
  17. Kim TJ, Lee EY, Kim YJ, Cho K-S, Ryu HW (2003) Degradation of polyaromatic hydrocarbons by Burkholderia cepacia 2A-12. World J Microbiol Biotechnol 19:411–417CrossRefGoogle Scholar
  18. Krivobok S, Kuony S, Meyer C, Louwagie M, Willison JC, Jouanneau Y (2003) Identification of pyrene-induced proteins in Mycobacterium sp. strain 6PY1: evidence for two ring-hydroxylating dioxygenase. J Bacteriol 185:3828–3841CrossRefGoogle Scholar
  19. Lee K, Park J-W, Ahn I-S (2003) Effect of additional carbon source on naphthalene biodegradation by Pseudomonas putida G7. J Hazard Mater 105:157–167CrossRefGoogle Scholar
  20. López Z, Vila J, Minguillón C, Grifoll M (2006) Metabolism of fluoranthene by Mycobacterium sp. strain AP1. Appl Microbiol Biotechnol 70:747–756CrossRefGoogle Scholar
  21. Lotfabad SK, Gray MR (2002) Kinetics of biodegradation of mixtures of polycyclic aromatic hydrocarbons. Appl Microbiol Biotechnol 60:361–365CrossRefGoogle Scholar
  22. Luan TG, Yu KSH, Zhong Y, Zhou HW, Lan CY, Tam NFY (2006) Study of metabolites from the degradation of polycyclic aromatic hydrocarbons (PAHs) by bacterial consortium enriched from mangrove sediments. Chemosphere 65:2289–2296CrossRefGoogle Scholar
  23. Molina M, Araujo R, Hodson RR (1999) Cross-induction of pyrene and phenanthrene in a Mycobacterium sp. isolated from polycyclic aromatic hydrocarbon contaminated river sediments. Can J Microbiol 45:520–529CrossRefGoogle Scholar
  24. Moody JD, Freeman JP, Doerge DR, Cerniglia CE (2001) Degradation of phenanthrene and anthracene by cell suspensions of Mycobacterium sp. strain PYR-1. Appl Environ Microbiol 67:1476–1483CrossRefGoogle Scholar
  25. Mueller JG, Chapman PJ, Blattmann BO, Pritchard PH (1990) Isolation and characterization of a fluoranthene-utilizing strain of Pseudomonas paucimobilis. Appl Environ Microbiol 56:1079–1086Google Scholar
  26. Nojiri HN, Nam J-W, Kosaka M, Morii K-I, Takemura T, Furihata K, Yamane H, Omori T (1999) Diverse oxygenations catalyzed by carbazole 1,9a-dioxygenase from Pseudomonas sp. strain CA10. J Bacteriol 181:3105–3113Google Scholar
  27. Perry JJ (1979) Microbial cooxidations involving hydrocarbons. Microbiol Rev 43:59–72Google Scholar
  28. Pinyakong O, Habe H, Supaka N, Pinpanichkarn P, Juntongjin K, Yoshida T, Furihata K, Nojiri H, Yamane H, Omori T (2000) Identification of novel metabolites in the degradation of phenanthrene by Sphingomonas sp. strain P2. FEMS Microbiol Lett 191:115–121CrossRefGoogle Scholar
  29. Pinyakong O, Habe H, Omori T (2003) The unique aromatic catabolic genes in sphingomonads degrading polycyclic aromatic hydrocarbons (PAHs). J Gen Appl Microbiol 49:1–19CrossRefGoogle Scholar
  30. Pinyakong O, Habe H, Kouzuma A, Nojiri H, Yamane H, Omori T (2004) Isolation and characterization of gene encoding polycyclic aromatic hydrocarbon dioxygenase from acenaphthene and acenaphthylene degrading Sphingomonas sp. strain A4. FEMS Microbiol Lett 238:297–305Google Scholar
  31. Ramsay MA, Swannell RPJ, Shipton WA, Duke NC, Hill RT (2000) Effect of bioremediation on the microbial community in oiled mangrove sediments. Mar Pollut Bull 41:413–419CrossRefGoogle Scholar
  32. Rehmann K, Hertkorn N, Kettrup AA (2001) Fluoranthene metabolism in Mycobacterium sp. strain KR20: identification of pathway intermediates during degradation and growth. Microbiology 147:2783–2794Google Scholar
  33. Röling WFM, Milner MG, Jones DM, Lee K, Daniel F, Swannell RJP, Head IM (2002) Robust hydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil spill bioremediation. Appl Environ Microbiol 68:5537–5548CrossRefGoogle Scholar
  34. Story SP, Parker SH, Kline JD, Tzeng T-RJ, Mueller JG, Kline EL (2000) Identification of four structural genes and two putative promoters necessary for utilization of naphthalene, phenanthrene, and fluoranthene by Sphingomonas paucimobilis var. EPA 505. Gene 260:155–169CrossRefGoogle Scholar
  35. Story SP, Kline EK, Hughes TA, Riley MB, Hayasaka SS (2004) Degradation of aromatic hydrocarbons by Sphingomonas paucimobilis strain EPA505. Arch Environ Contam Toxicol 47:168–176CrossRefGoogle Scholar
  36. Stringfellow WT, Aitken MD (1995) Competitive metabolism of naphthalene, methylnaphthalenes, and fluorene by phenanthrene-degrading Pseudomonads. Appl Environ Microbiol 61:357–362Google Scholar
  37. Swannell RP, Lee K, McDonagh M (1996) Field evaluations of marine oil spill bioremediation. Microbiol Rev 60:342–365Google Scholar
  38. Tam NFY (2006) Pollution studies on mangroves in Hong Kong and mainland China. In: Wolanski E (ed) The environment in Asia Pacific harbours. Springer, Berlin Heidelberg New York, pp 147–163CrossRefGoogle Scholar
  39. Tam NFY, Ke L, Wang XH, Wong YS (2001) Contamination of polycyclic aromatic hydrocarbons in surface sediments of mangrove swamps. Environ Pollut 114:252–263Google Scholar
  40. Tam NFY, Guo CL, Yau WY, Wong YS (2002) Preliminary study on biodegradation of phenanthrene by bacteria isolated from mangrove sediments in Hong Kong. Mar Pollut Bull 45:316–324CrossRefGoogle Scholar
  41. Tongpim S, Pickard MA (1999) Cometabolic oxidation of phenanthrene to phenanthrene tran-9,10-dihydrodiol by Mycobacterium strain S1 growing on anthracene in the presence of phenanthrene. Can J Microbiol 45:369–376CrossRefGoogle Scholar
  42. van Herwijnen R, Wattiau P, Bastiaens L, Daal L, Jonker L, Springael D, Govers HAJ, Parsons JR (2003) Elucidation of the metabolic pathway of fluorene and cometabolic pathways of phenanthrene, fluoranthene, anthracene and dibenzothiophene by Sphingomonas sp. LB126. Res Microbiol 154:199–206CrossRefGoogle Scholar
  43. Wattiau P, Bastiaens L, van Herwijnen R, Daal L, Parsons JR, Renard M-E, Springael D, Cornelis GR (2001) Fluorene degradation by Sphingomonas sp. LB126 proceeds through protocatechuic acid: a genetic analysis. Res Microbiol 152:861–872CrossRefGoogle Scholar
  44. Yamazoe A, Yagi O, Oyaizu H (2004) Biotransformation of fluorene, diphenyl ether, dibenzo-p-dioxin and carbazole by Janibacter sp. Biotechnol Lett 26:479–486CrossRefGoogle Scholar
  45. Zhong Y, Luan TG, Zhou HW, Lan CY, Tam NFY (2006) Metabolite production in degradation of pyrene alone or in a mixture with another polycyclic aromatic hydrocarbon by Mycobacterium sp. Environ Toxicol Chem 25:2853–2859CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.State Key Laboratory of Biocontrol, School of Life SciencesSun Yat-Sen UniversityGuangzhouPeople’s Republic of China
  2. 2.Futian-CityU Mangrove Research & Development CentreFutian National Nature ReserveShenzhenPeople’s Republic of China
  3. 3.Department of Biology and ChemistryCity University of Hong KongKowloonPeople’s Republic of China

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