Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Biodegradation of dimethyl phthalate by Sphingomonas sp. isolated from phthalic-acid-degrading aerobic granules

  • 344 Accesses

  • 19 Citations


Phthalic acid esters (PAEs) contamination in water, air, and soil is one of the major environmental concerns in many countries. Besides the PAE biodegradation process, the PAE degrading bacteria have become one of the focuses of study. This study reports the successful isolation of one kind of indigenous bacterium PA-02 from phthalic acid (PA)-degrading aerobic granules. Based on its 16S ribosomal DNA sequence, isolate PA-02 was identified as Sphingomonas genus with 100% similarity to Sphingomonas sp. strain D84532. Strain PA-02 was a Gram-negative, rod-shaped bacterium with strong auto-aggregation ability. In particular, the strain PA-02 possessed PAE-degrading ability without acclimation. Results of growth tests showed that strain PA-02 could degrade dimethyl phthalate (DMP), dibutyl phthalate, and diethylhexyl phthalate. The specific degradation rates of DMP and PA were concentration-dependent with maximum values of 0.4 g-DMP g−1 biomass h−1 and 1.3 g-PA g−1 biomass h−1, respectively. Kinetic studies also revealed that PA-02 was robust under high concentrations of DMP and PA. Even when the PA concentration was increased to 1,000.0 mg l−1, the specific PA degradation rate was about 0.25 g-PA g−1 biomass h−1. The corresponding value for DMP was 0.067 g-DMP g−1 biomass h−1 at 1,000 mg l−1.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. Bauer MJ, Herrmann R (1997) Estimation of the environmental contamination by phthalic acid esters leaching from household wastes. Sci Total Environ 208:49–57

  2. Boone DR, Brenner DJ, Krieg NR, Staley JT (2005) Bergey’s manual of systematic bacteriology, 2nd edn, vol 2. Springer, East Lansing, USA

  3. Bramucci M, Singh M, Nagarajan V (2002) Biotransformation of p-xylene and 2,6-dimethylnaphthalene by xylene monooxygenase cloned from a Sphingomonas isolate. Appl Environ Microbiol 59:679–684

  4. Chang BV, Yang CM, Cheng CH, Yuan SY (2004) Biodegradation of phthalate esters by two bacteria strains. Chemosphere 55:533–538

  5. Chatterjee S, Dutta TK (2003) Metabolism of butyl benzyl phthalate by Gordonia sp. strain MTCC 4818. Biochem Biophys Res Commun 309:36–43

  6. Chen JA, Li X, Li J, Cao J, Qiu ZQ, Zhao Q, Xu C, Shu WQ (2007) Degradation of environmental endocrine disruptor di-2-ethylhexyl phthalate by a newly discovered bacterium, Microbacterium sp strain CQ0110Y. Appl Microbiol Biotechnol 74:676–682

  7. Colon I, Caro D, Bourdony CJ, Rosario O (2000) Identification of phthalate esters in the serum of young Puerto Rican girls with premature breast development. Environ Health Perspect 108:895–900

  8. Fan YZ, Cheng SP, Gu JD (2001) Degradation of phthalic acid and dimethyl phthalate ester by enrichment cultures of aerobic bacteria. Paper presented at the Conference proceedings of IWA Asia Environmental Technology 2001. Singapore, pp 547–554

  9. Ivanov V, Wang XH, Tay STL, Tay JH (2006) Bioaugmentation and enhanced formation of microbial granules used in aerobic wastewater treatment. Appl Microbiol Biotechnol 70:374–381

  10. Jiang HL, Tay JH, Maszenan AM, Tay STL (2004) Bacterial diversity and function of aerobic granules engineered in a sequencing batch reactor for phenol degradation. Appl Environ Microbiol 70:6767–6775

  11. Kleerebezem R, Pol LWH, Lettinga G (1999) Anaerobic degradation of phthalate isomers by methanogenic consortia. Appl Environ Microbiol 65:1152–1160

  12. Knudsen FR, Pottinger TG (1999) Interaction of endocrine disrupting chemicals, singly and in combination, with estrogen-, androgen-, and corticosteroid-binding sites in rainbow trout (Oncorhynchus mykiss). Aquat Toxicol 44:159–170

  13. Lane DJ (1991) 16S/23S rRNA sequencing. In: Goodfellow ESM (ed) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, England, pp 115–175

  14. Leys NM, Ryngaert A, Bastiaens L, Top EM, Verstraete W, Springael D (2005) Culture independent detection of Sphingomonas sp EPA 505 related strains in soils contaminated with polycyclic aromatic hydrocarbons (PAHs). Microb Ecol 49:443–450

  15. Liang DW, Zhang T, Fang HHP (2007) Denitrifying degradation of dimethyl phthalate. Appl Microbiol Biotechnol 74:221–229

  16. Liu YS, Zhang J, Zhang ZZ (2004) Isolation and characterization of polycyclic aromatic hydrocarbons-degrading Sphingomonas sp. strain ZL5. Biodegradation 15:205–212

  17. Marttinen SK, Kettunen RH, Sormunen KM, Rintala JA (2003) Removal of bis(2-ethylhexyl) phthalate at a sewage treatment plant. Water Res 37:1385–1393

  18. Miyachi N, Tanaka T, Suzuki T, Hotta Y, Oori T (1993) Microbial oxidation of dimethylnaphthalene isomers. Appl Environ Microbiol 59:1504–1506

  19. Neefs JM, Vandepeer Y, Hendriks L, Wachter R (1990) Compilation of small ribosomal-subunit RNA sequences. Nucleic Acids Res 18:2237–2317

  20. Nielsen E, Larsen PB (1996) Toxicological evaluation and limit values for DEHP and phthalates other than DEHP. Danish Environmental Protection Agency

  21. Nishio E, Yoshikawa H, Wakayama M, Tamura H, Morita S, Tomita Y (2005) Isolation and identification of Sphingomonas sp. that yields tert-octylphenol monoethoxylate under aerobic conditions. Biosci Biotechnol Biochem 69:1226–1231

  22. Nomura Y, Takada N, Oshima Y (1989) Isolation and identification of phthalate-utilization bacteria. J Ferment Bioeng 67:297–299

  23. Nomura Y, Nakagawa M, Ogawa N, Harashima S, Oshima Y (1992) Genes in PHT plasmid encoding the initial degradation pathway of phthalate in Pseudomonas-putida. J Ferment Bioeng 74:333–344

  24. Ono K, Nozaki M, Hayaishi O (1970) Purification and some properties of protocatechuate 4,5-dioxgenase. Biochim Biophys Acta 220:224–238

  25. Parker WJ, Monteith HD, Melcer H (1994) Estimation of anaerobic biodegradation rates for toxic organic compounds in municipal sludge digestion. Water Res 28:1779–1789

  26. Petrovilc M, Eljarrat E, Liopez MJ, Barcelio D (2001) Analysis and environmental levels of endocrine-disrupting compounds in freshwater sediments. TRAC-Trends Anal Chem 20:637–648

  27. Roslev P, Vorkamp K, Aarup J, Frederiksen K, Nielsen PH (2007) Degradation of phthalate esters in an activated sludge wastewater treatment plant. Water Res 41:969–976

  28. Sasaki M, Maki J, Oshiman K, Matsumura Y, Tsuchido T (2005) Biodegradation of bisphenol A by cells and cell lysate from Sphingomonas sp. strain AO1. Biodegradation 16:449–459

  29. Scholz N, Diefenbach R, Rademacher I, Linneman D (1997) Biodegradation of DEHP, DBP, and DINP: poorly water soluble and widely used phthalate plasticizers. Bull Environ Contam Toxicol 58:527–534

  30. Shelton DR, Boyd SA, Tiedje JM (1984) Anaerobic biodegradation of phthalic acid esters in sludge. Environ Sci Technol 18:93–97

  31. Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray RGE, Wood WA, Krieg NR (eds) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, DC, pp 607–654

  32. Staples CA, Peterson DR, Parkerton TF, Adams WJ (1997) The environmental fate of phthalate esters: a literature review. Chemosphere 35:667–749

  33. Takeuchi M, Sawada H, Oyaizu H, Yokota A (1994) Phylogenetic evidence for Sphingomonas and Rhizomonas as nonphotosynthetic members of the alpha-4 subclass of the Proteobacteria. Int J Syst Bacteriol 44:308–314

  34. Tay STL, Hemond HF, Polz MF, Cavanaugh CM, Dejesus I, Krumholz LR (1998) Two new Mycobacterium strains and their role in toluene degradation in a contaminated stream. Appl Environ Microbiol 64:1715–1720

  35. Tay STL, Ivanov V, Yi S, Zhuang WQ, Tay JH (2002) Presence of anaerobic bacteroides in aerobically grown microbial granules. Microb Ecol 44:278–285

  36. Tay STL, Jiang HL, Tay JH (2004) Functional analysis of microbial community in phenol-degrading aerobic granules cultivated in SBR. Water Sci Technol 50:229–234

  37. US EPA (1992 and update) Code of federal regulations, 40 CFR, Part 136

  38. Vamsee-Krishna C, Mohan Y, Phale PS (2006) Biodegradation of phthalate isomers by Pseudomonas aeruginosa PP4, Pseudomonas sp PPD and Acinetobacter lwoffii ISP4. Appl Microbiol Biotechnol 72:1263–1269

  39. Vega D, Bastide J (2003) Dimethylphthalate hydrolysis by specific microbial esterase. Chemosphere 51:663–668

  40. Wang JL, Liu P, Qian Y (1995) Microbial degradation of di-n-butyl phthalate. Chemosphere 31:4051–4056

  41. Wang JL, Liu P, Qian Y (1997) Biodegradation of phthalic acid esters by immobilized microbial cells. Environ Int 23:775–782

  42. Wang JL, Liu P, Shi HC, Qian Y (1998) Kinetics of biodegradation of di-n-butyl phthalate in continuous culture system. Chemosphere 37:257–264

  43. Wang YY, Fan YZ, Gu JD (2003) Aerobic degradation of phthalic acid by Comamonas acidovoran Fy-1 and dimethyl phthalate ester by two reconstituted consortia from sewage sludge at high concentrations. World J Microbiol Biotechnol 19:811–815

  44. Yurkov V, Stackebrandt E, Buss O, Vermeglio A, Gorlenko V, Beatty JT (1997) Reorganization of the genus Erythromicrobium: description of “Erythromicrobium sibiricum” as Sandaracinobacter sibiricus gen. nov., sp. nov., and of “Erythromicrobium ursincola” as Erythromonas ursincola gen. nov., sp. nov. Int J Syst Bacteriol 47:1172–1178

  45. Zeng F, Cui KY, Li XD, Fu JM, Sheng GY (2004) Biodegradation kinetics of phthalate esters by Pseudomonas fluoresences FS1. Process Biochem 39:1125–1129

  46. Zhang G, Peardon KF (1990) Parametric study of diethyl phthalate biodegradation. Biotechnol Lett 12:699–704

  47. Ziogou K, Kirk PWW, Lester JN (1989) Behavior of phthalic-acid esters during batch anaerobic-digestion of sludge. Water Res 23:743–748

Download references


We thank Mr. C.L. Cheah for correcting the language.

Author information

Correspondence to Ping Zeng.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zeng, P., Moy, B.Y., Song, Y. et al. Biodegradation of dimethyl phthalate by Sphingomonas sp. isolated from phthalic-acid-degrading aerobic granules. Appl Microbiol Biotechnol 80, 899–905 (2008). https://doi.org/10.1007/s00253-008-1632-x

Download citation


  • PAEs degradation
  • Dimethyl phthalate
  • 16S rDNA
  • Sphingomonas
  • Kinetics