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

Single-culture aerobic granules with Acinetobacter calcoaceticus

  • 338 Accesses

  • 33 Citations


Aerobic granules are cultivated by a single bacterial strain, Acinetobacter calcoaceticus, in a sequencing batch reactor (SBR). This strain presents as a good phenol reducer and an efficient auto coagulator in the presence of phenol, mediated by heat-sensitive adhesins proteins. Stable 2.3-mm granules were formed in the SBR following a 7-week cultivation. These granules exhibit excellent settling attributes and degrade phenol efficiently at concentrations of 250–2,000 mg l−1. The corresponding phenol degradation rate reached 993.6 mg phenol g−1 volatile suspended solids (VSS) day−1 at 250 mg l−1 phenol and 519.3 mg phenol g−1 VSS day−1 at 2,000 mg l−1 phenol concentration. Meanwhile, free A. calcoaceticus cells were fully inhibited at phenol >1,500 mg l−1. Denaturing gradient gel electrophoresis fingerprint profile demonstrated no genetic modification in the strain during aerobic granulation. The present single-strain granules showed long-term structural stability and performed high phenol degrading capacity and high phenol tolerance. The confocal laser scanning microscopic test revealed that live A. calcoaceticus cells principally distributed at 200–250 μm beneath the outer surface, with an extracellular polymeric substance layer covering them to defend phenol toxicity. Autoaggregation assay tests demonstrated the possibly significant role of secreted proteins on the formation of single-culture A. calcoaceticus granules.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Adav SS, Lee DJ (2007) Extraction of extracellular polymeric substances from aerobic granule with compact interior structure. J Hazard Mater. DOI https://doi.org/10.1016/j.jhazmat.2007.11.058

  2. Adav SS, Chen MY, Lee DJ, Ren NQ (2007a) Degradation of phenol by Acinetobactor strain isolated from aerobic granules. Chemosphere 67:1566–1572

  3. Adav SS, Chen MY, Lee DJ, Ren NQ (2007b) Degradation of phenol by aerobic granules and isolated yeast Candida tropicalis. Biotechnol Bioeng 96:844–852

  4. Adav SS, Lee DJ, Ren NQ (2007c) Biodegradation of pyridine using aerobic granules in the presence of phenol. Water Res 41:2903–2910

  5. APHA (1998) The standard methods for the examination of water and wastewater, , 20th edn. American Public Health Association, Washington, DC

  6. Beun JJ, Hendriks A, van Loosdrecht MCM, Morgenroth E, Wilderer PA, Heijnen JJ (1999) Aerobic granulation in a sequencing batch reactor. Water Res 33:2283–2290

  7. Buchan L (1983) Possible biological mechanism of phosphorus removal. Water Sci Technol 15:197–217

  8. Chen MY, Lee DJ, Tay JH, Show KY (2007) Staining of extracellular polymeric substances and cells in bio-aggregates. Appl Microbiol Biotechnol 75:467–474

  9. Frolund B, Palmgren R, Keiding K, Nielsen PH (1996) Extraction of extracellular polymers from activated sludge using a cation exchange resin. Water Res 30:1749–1758

  10. Gaoa Y, Yu XZ, Wu SC, Cheung KC, Tamb NFY, Qian PY, Wong MH (2006) Interactions of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) on enhanced dissipation of spiked phenanthrene and pyrene in waterlogged soil. Sci Total Environ 372:1–11

  11. Gaudy AF (1962) Colorimetric determination of protein and carbohydrate. Ind Water Wastes 7:17–22

  12. Hao OJ, Kim MH, Seagren EA, Kim H (2002) Kinetics of phenol and chlorophenol utilization by Acinetobacter species. Chemosphere 46:797–807

  13. Ishii S, Koki J, Unno H, Hori K (2004) Two morphological types of cell appendages on a strongly adhesive bacterium, Acinetobacter sp. strain Tol 5. Appl Environ Microbiol 70:5026–5029

  14. 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

  15. Jiang HL, Tay JH, Maszenan AM, Tay STL (2006a) Enhanced phenol biodegradation and aerobic granulation by two coaggregating bacterial strains. Environ Sci Technol 40:6137–6142

  16. Jiang HL, Tay STL, Maszenan AM, Tay JH (2006b) Physiological traits of bacterial strains isolated from phenol-degrading aerobic granules. FEMS Microbiol Ecol 57:182–191

  17. Jiang HL, Maszenan AM, Tay JH (2007) Bioaugmentation of coexistence of two functionally similar bacterial strains in aerobic granules. Appl Microbiol Biotechnol 75:1191–1200

  18. Kim MH, Hao OJ, Wang NS (1997) Acinetobacter isolates from different activated sludge processes: characteristics and neural network identification. FEMS Microbiol Ecol 23:217–227

  19. Lotter LH (1985) The role of bacterial phosphate metabolism in enhanced phosphate removal from the activated sludge process. Water Sci Technol 17:127–138

  20. Peng DC, Bernet N, Delgenes JP, Moletta R (1999) Aerobic granular sludge—a case report. Water Res 33:890–893

  21. Su KZ, Yu HQ (2005) Formation and characterization of aerobic granules in a sequencing batch reactor treating soybean-processing wastewater. Environ Sci Technol 39:2818–2828

  22. Tay JH, Liu QS, Liu Y (2001) Microscopic observation of aerobic granulation in sequential aerobic sludge blanket reactor. J Appl Microbiol 91:168–175

  23. 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

  24. Watanabe K, Hino S, Takahashi N (1996) Responses of activated sludge to an increase in phenol loading. J Ferment Bioeng 82:522–524

Download references

Author information

Correspondence to Duu-Jong Lee.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Adav, S.S., Lee, D. Single-culture aerobic granules with Acinetobacter calcoaceticus . Appl Microbiol Biotechnol 78, 551–557 (2008). https://doi.org/10.1007/s00253-007-1325-x

Download citation


  • Aerobic granule
  • Single culture
  • Autoaggregation
  • Acinetobactor calcoaceticus