Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Biodegradation of methyl t-butyl ether by aerobic granules under a cosubstrate condition

  • 203 Accesses

  • 19 Citations


Aerobic granules efficient at degrading methyl tert-butyl ether (MTBE) with ethanol as a cosubstrate were successfully developed in a well-mixed sequencing batch reactor (SBR). Aerobic granules were first observed about 100 days after reactor startup. Treatment efficiency of MTBE in the reactor during stable operation exceeded 99.9%, and effluent MTBE was in the range of 15–50 μg/L. The specific MTBE degradation rate was observed to increase with increasing MTBE initial concentration from 25 to 500 mg/L, which peaked at 22.7 mg MTBE/g (volatile suspended solids)·h and declined with further increases in MTBE concentration as substrate inhibition effects became significant. Microbial-community deoxyribonucleic acid profiling was carried out using denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S ribosomal ribonucleic acid. The reactor was found to be inhabited by several diverse bacterial species, most notably microorganisms related to the genera Sphingomonas, Methylobacterium, and Hyphomicrobium vulgare. These organisms were previously reported to be associated with MTBE biodegradation. A majority of the bands in the reactor represented a group of organisms belonging to the FlavobacteriaProteobacteria–Actinobacteridae class of bacteria. This study demonstrates that MTBE can be effectively degraded by aerobic granules under a cosubstrate condition and gives insight into the microorganisms potentially involved in the process.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Bhatti ZI, Toda H, Furukawa K (2002) p-Nitrophenol degradation by activated sludge attached on nonwovens. Water Res 36:1135–1142

  2. Cottrell MT, Kirchman DL (2000) Natural assemblages of marine proteobacteria and members of the CyophagaFlavobacter cluster consuming low and high molecular-weight dissolved organic matter. Appl Environ Microbiol 66:1692–1697

  3. De Kreuk MK, Picioreanu C, Hosscini M, Xavier JB, van Loosdrecht MCM (2007) Kinetic model of a granular sludge SBR: influence on nutrient removal. Biotechnol Bioeng 97:801–815

  4. Deeb RA, Scow KM, Cohen LA (2000) Aerobic MTBE biodegradation: an examination of past studies, current challenges and future research directions. Biodegradation 11:171–186

  5. Depasquier D, Revah S, Auria R (2002) Biofiltration of methyl tert-butyl ether vapors by cometabolism with pentane: modeling and experimental approach. Environ Sci Technol 36:247–253

  6. Dong MX, Ma Z, Chang K (2007) MTBE production technology and market prospect analyse. Petrochem Industry Appl 26:6–9

  7. Fortin NY, Deshusses MA (1999) Treatment of methyl tert-butyl ether vapors in biotrickling filters. 1. reactor startup, steady-state performance, and culture characteristics. Environ Sci Technol 33:2980–2986

  8. Fortin NY, Morales M, Nakagawa Y, Focht DD, Deshusses MA (2001) Methyl tert-butyl ether (MTBE) degradation by a microbial consortium. Environ Microbiol 3:407–416

  9. François A, Mathis H, Godefroy D, Piveteau P, Fayolle F, Monot F (2002) Biodegradation of methyl tert-butyl ether and other fuel oxygenates by a new strain, Mycobacterium austroafricanum IFP 2012. Appl Environ Microbiol 68:2754–2762

  10. Henriques DSI, Holbrook RD, Kelly RT II, Love NG (2005) The impact of floc size on respiration inhibition by soluble toxicants—a comparative investigation. Water Res 39:2559–2568

  11. Jiang HL, Tay JH, Tay STL (2002) Aggregation of immobilized activated sludge cells into aerobically grown microbial granules for the aerobic biodegradation of phenol. Lett Appl Microbiol 35:439–445

  12. Jiang HL, Tay JH, Liu Y, 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

  13. Kelley ST, Theisen U, Angenent LT, Amand AS, Pace NR (2004) Molecular analysis of shower curtain biofilm microbes. Appl Environ Microbiol 70:4187–4192

  14. Kowalchuk GA, Stephen JR, DeBoer W, Prosser JI, Embley TM, Woldendorp JW (1997) Analysis of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in coastal sand dunes by denaturing gradient gel eletrophoresis and sequencing of PCR-amplified 16S ribosomal DNA fragments. Appl Environ Microbiol 63:1489–1497

  15. Lin CW, Lin NC, Liu MC (2007) Biodegradation kinetics and microbial communities associated with methyl tert-butyl ether removal in a biotrickling filter. Chem Eng J 127:143–149

  16. Liu Y, Tay JH (2004) State of the art of biogranulation technology for wastewater treatment. Biotechnol Adv 22:533–563

  17. Mo K, Lora CO, Wanken AE, Javanmardian M, Yang X, Kulpa CF (1997) Biodegradation of methyl tert-bulty ether by pure bacterial cultures. Appl Mirobiol Biotechnol 47:69–72

  18. MØller S, Sternberg C, Andersen JB, Christensen BB, Ramos JL, Givskov M, Molin S (1998) In situ gene expression in mixed-culture biofilms: evidence of metabolic interactions between community members. Appl Environ Microbiol 64:721–732

  19. Morrison JR, Suidan MT, Venosa AD (2002) Use of membrane bioreactor for biodegradation of MTBE in contaminated water. J Environ Eng 128:836–841

  20. Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700

  21. Nikitin DI, Slabova OI, Pitryuk IA, Sorokin VV, Oranskaya MS (1998) Response of psychoactive oligotrophic bacteria with an unusual lipid composition to cultivation temperatures. Microbiology 67(1):54–60

  22. Piveteau P, Fayolle F, Vandecasteele JP, Monot F (2001) Biodegradation of tert-butyl alcohol and related xenobiotics by a methylotrophic bacterial isolate. Appl Microbial Biotechnol 55:369–373

  23. Pruden A, Suidan MT, Venosa AD, Wilson GJ (2001) Biodegradation of methy tert-butyl ether under various substrate conditions. Envrion Sci Technol 35:4235–4241

  24. Salanitro JP, Diaz MP, Williams MP, Wisniewski HL (1994) Isolation of a bacterial culture that degrades methyl t-butyl ether. Appl Environ Microbiol 60:2593–2596

  25. Sedran MA, Pruden A, Wilson GJ, Suidan MT, Venosa AD (2002) Effect of BTEX on degradation of MTBE and TBA by mixed bacterial consortium. J Environ Eng 128:830–832

  26. Su KZ, Yu HQ (2005) Formation and characterization of aerobic granules in a sequencing batch reactor treating soybean-processing wastewater. Environ Sci Technol 39:2818–2827

  27. Tay STL, Moy BYP, Jiang HL, Tay JH (2005) Rapid cultivation of stable aerobic phenol-degrading granules using acetate-fed granules as microbial seed. J Biotechnol 115:387–395

  28. Van Aken B, Yoon JM, Schnoor JL (2004) Biodegradation of nitrosubstituted explosives 2,4,6-trinitrotoluene, hexahydro-1,3,5-trinitro-1,3,5-triazine, an octahydro-1,3,5,7-tetranitro-1,3,5,-tetrazocine by a phytosymbiotic Methylobacterium sp associated with poplar tissues (Populus deltoids x nigra DN 34). Appl Environ Microbiol 70:508–517

  29. Wilson GJ, Pruden A, Suidan MT, Venosa AD (2002) Biodegradation kinetics of MTBE in laboratory batch and continuous flow reactors. J Environ Eng 128:824–829

  30. Yi S, Zhuang WQ, Wu B, Tay STL, Tay JH (2006) Biodegradation of p-Nitrophenol by aerobic granules in a sequencing batch reactor. Environ Sci Technol 40:2396–2401

  31. Zein MM, Suidan MT, Venosa AD (2004) MtBE biodegradation in a gravity flow, high-biomass retaining bioreactor. Environ Sci Technol 38:3449–3456

  32. Zhang LL, Zhang B, Huang YF, Cai WM (2005) Application of aerobic granulation in polishing the UASB effluent. Environ Technol 26:1327–1334

  33. Zhuang WQ, Tay JH, Maszenan AM, Tay STL (2002) Bacillus naphthovorans sp nov from oil-contaminated tropical marine sediments and its role in naphthalene biodegradation. Appl Microbiol Biotechnol 58:547–553

Download references


The authors wish to thank the Natural Science Foundation of China (Grant no. 20476099) and the Natural Science Foundation of Zhejiang Province (Grant no. Y507270) for the support of this study.

Author information

Correspondence to J. M. Chen.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zhang, L.L., Chen, J.M. & Fang, F. Biodegradation of methyl t-butyl ether by aerobic granules under a cosubstrate condition. Appl Microbiol Biotechnol 78, 543–550 (2008).

Download citation


  • Aerobic granule
  • MTBE
  • DGGE
  • Microbial community