Extraction of structural extracellular polymeric substances from aerobic granular sludge

Simon Felz, Salah Al-Zuhairy, Olav Andreas Aarstad, Mark C M van Loosdrecht, Y. Lin

Research output: Contribution to journalArticleScientificpeer-review

170 Citations (Scopus)
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To evaluate and develop methodologies for the extraction of gel-forming extracellular polymeric substances (EPS), EPS from aerobic granular sludge (AGS) was extracted using six different methods (centrifugation, sonication, ethylenediaminetetraacetic acid (EDTA), formamide with sodium hydroxide (NaOH), formaldehyde with NaOH and sodium carbonate (Na2CO3) with heat and constant mixing). AGS was collected from a pilot wastewater treatment reactor. The ionic gel-forming property of the extracted EPS of the six different extraction methods was tested with calcium ions (Ca2+). From the six extraction methods used, only the Na2CO3 extraction could solubilize the hydrogel matrix of AGS. The alginatelike extracellular polymers (ALE) recovered with this method formed ionic gel beads with Ca2+. The Ca2+-ALE beads were stable in EDTA, formamide with NaOH and formaldehyde with NaOH, indicating that ALE are one part of the structural polymers in EPS. It is recommended to use an extraction method that combines physical and chemical treatment to solubilize AGS and extract structural EPS.

Original languageEnglish
Article numbere54534
JournalJournal of Visualized Experiments
Issue number115
Publication statusPublished - 26 Sept 2016


  • Aerobic granular sludge
  • Alginate-like extracellular polymers (ALE)
  • Biofilm
  • Biofouling
  • Environmental sciences
  • Extracellular polymeric substances (EPS)
  • Extraction
  • Functional polymer
  • Hydrogel
  • Structural polymer


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