Applied Microbiology and Biotechnology

, Volume 85, Issue 5, pp 1589–1599 | Cite as

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

  • Paul D’Abzac
  • François Bordas
  • Eric Van Hullebusch
  • Piet N. L. Lens
  • Gilles GuibaudEmail author
Environmental Biotechnology


The characteristics of the extracellular polymeric substances (EPS) extracted with nine different extraction protocols from four different types of anaerobic granular sludge were studied. The efficiency of four physical (sonication, heating, cationic exchange resin (CER), and CER associated with sonication) and four chemical (ethylenediaminetetraacetic acid, ethanol, formaldehyde combined with heating, or NaOH) EPS extraction methods was compared to a control extraction protocols (i.e., centrifugation). The nucleic acid content and the protein/polysaccharide ratio of the EPS extracted show that the extraction does not induce abnormal cellular lysis. Chemical extraction protocols give the highest EPS extraction yields (calculated by the mass ratio between sludges and EPS dry weight (DW)). Infrared analyses as well as an extraction yield over 100% or organic carbon content over 1 g g−1 of DW revealed, nevertheless, a carry-over of the chemical extractants into the EPS extracts. The EPS of the anaerobic granular sludges investigated are predominantly composed of humic-like substances, proteins, and polysaccharides. The EPS content in each biochemical compound varies depending on the sludge type and extraction technique used. Some extraction techniques lead to a slightly preferential extraction of some EPS compounds, e.g., CER gives a higher protein yield.


Anaerobic granular sludge Extracellular polymeric substances (EPS) Extraction Biochemical composition 



This study was supported by a research grant provided by the Conseil Regional du Limousin. The authors thank Audrey Asmond for her technical assistance in EPS extraction and characterization.


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Paul D’Abzac
    • 1
  • François Bordas
    • 1
  • Eric Van Hullebusch
    • 2
  • Piet N. L. Lens
    • 3
  • Gilles Guibaud
    • 1
    Email author
  1. 1.Groupement de Recherche Eau Sol EnvironnementUniversité de LimogesLimogesFrance
  2. 2.Laboratoire Géomatériaux et EnvironnementUniversité Paris-EstMarne la Vallée Cedex 2France
  3. 3.Sub-department of Environmental TechnologyWageningen UniversityWageningenThe Netherlands

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