Advertisement

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

Metal binding properties of extracellular polymeric substances extracted from anaerobic granular sludges

Abstract

Extracellular polymeric substances (EPS) were extracted from four anaerobic granular sludges with different procedures to study their involvement in biosorption of metallic elements. EPS extracts are composed of closely associated organic and mineral fractions. The EPS macromolecules (proteins, polysaccharides, humic-like substances, nucleic, and uronic acids) have functional groups potentially available for the binding of metallic elements. The acidic constants of these ionizable groups are: pK a1 (4–5) corresponding to the carboxyl groups; pK a2 (6–7) corresponding to the phosphoric groups; pK a3 (8–10) and pK a4 (≈10) corresponding to the phenolic, hydroxyl, and amino groups. The polarographic study confirms the higher affinity of the EPS to bind to lead than to cadmium. Moreover, the binding of these metallic compounds with the EPS is a mix of several sorption mechanisms including surface complexation, ion exchange, and flocculation. Inorganic elements were found as ions linked to organic molecules or as solid particles. The mineral fraction affects the binding properties of the EPS, as the presence of salts decreases the EPS binding ability. Calcite and apatite particles observed on SEM images of EPS extracts can also sorb metallic elements through ion exchange or surface complexation.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  1. Al-Degs YS, El-Barghouthi MI, Issa AA, Khraisheh MA, Walker GM (2006) Sorption of Zn(II), Pb(II), and Co(II) using natural sorbents: equilibrium and kinetic studies. Water Res 40:2645–2658

  2. Bhaskar PV, Bhosle NB (2006) Bacterial extracellular polymeric substance (EPS): a carrier of heavy metals in the marine food-chain. Environ Int 32:191–198

  3. Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54:484–489

  4. Braissant O, Decho AW, Dupraz C, Glunk C, Przekop KM, Visscher PT (2007) Exopolymeric substances of sulfate-reducing bacteria: interactions with calcium at alkaline pH and implication for formation of carbonate minerals. Geobiology 5:401–411

  5. Burnett PGG, Daughney CJ, Peak D (2006) Cd adsorption onto Anoxybacillus flavithermus: surface complexation modeling and spectroscopic investigations. Geochim Cosmochim Acta 70:5253–5269

  6. Burton K (1956) A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of desoxyribonucleic acid. Biochem J 62:315–323

  7. Chai X, Takayuki S, Cao X, Guo Q, Zhao Y (2007) Spectroscopic studies of the progress of humification processes in humic substances extracted from refuse in a landfill. Chemosphere 69:1446–1453

  8. Comte S, Guibaud G, Baudu M (2006a) Relations between extraction protocols for activated sludge extracellular polymeric substances (EPS) and complexation properties of Pb and Cd with EPS: part II. Consequences of EPS extraction methods on Pb2+ and Cd2+ complexation. Enzym Microb Technol 38:246–252

  9. Comte S, Guibaud G, Baudu M (2006b) Relations between extraction protocols for activated sludge extracellular polymeric substances (EPS) and EPS complexation properties: part I. Comparison of the efficiency of eight EPS extraction methods. Enzym Microb Technol 38:237–245

  10. Cox JS, Smith DS, Warren LA, Ferris FG (1999) Characterizing heterogeneous bacterial surface functional groups using discrete affinity spectra for proton binding. Environ Sci Technol 33:4514–4521

  11. d'Abzac P, Bordas F, Joussein E, Van Hullebusch E, Lens PNL, Guibaud G (2010a) Characterization of the mineral fraction associated to extracellular polymeric substances (EPS) in anaerobic granular sludges. Environ Sci Technol 44:412–418

  12. d'Abzac P, Bordas F, Van Hullebusch E, Lens PNL, Guibaud G (2010b) Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols. Appl Microbiol Biotechnol 85:1589–1599

  13. d'Abzac P, Bordas F, van Hullebusch E, Lens PNL, Guibaud G (2010c) Effects of extraction procedures on metal binding properties of extracellular polymeric substances (EPS) from anaerobic granular sludges. Colloids Surf B: Biointerfaces 80:161–168

  14. Dittrich M, Sibler S (2005) Cell surface groups of two picocyanobacteria strains studied by zeta potential investigations, potentiometric titration, and infrared spectroscopy. J Colloid Interface Sci 286:487–495

  15. Donlan RM (2002) Biofilms: microbial life on surfaces. Emerg Infect Dis 8:881–890

  16. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

  17. Dupraz C, Reid RP, Braissant O, Decho AW, Norman RS, Visscher PT (2009) Processes of carbonate precipitation in modern microbial mats. Earth Sci Rev 96:141–162

  18. Eboigbodin KE, Biggs CA (2008) Characterization of the extracellular polymeric substances produced by Escherichia coli using infrared spectroscopic, proteomic, and aggregation studies. Biomacromol 9:686–695

  19. Flemming HC, Wingender J (2001) Relevance of microbial extracellular polymeric substances (EPSs)—part I: structural and ecological aspects. Water Sci Technol 43:1–8

  20. Frings CS, Dunn RT (1970) A colorimetric method for determination of total serum lipids based on the sulfo-phospho-vanillin reaction. Am J Clin Pathol 53:89–91

  21. Frolund B, Griebe T, Nielsen PH (1995) Enzymatic activity in the activated-sludge floc matrix. Appl Microbiol Biotechnol 43:755–761

  22. Giannakopoulos E, Christoforidis KC, Tsipis A, Jerzykiewicz M, Deligiannakis Y (2005) Influence of Pb(II) on the radical properties of humic substances and model compounds. J Phys Chem A 109:2223–2232

  23. Giannakopoulos E, Drosos M, Deligiannakis Y (2009) A humic-acid-like polycondensate produced with no use of catalyst. J Colloid Interface Sci 336:59–66

  24. Gonzalez-Gil G, Lens PNL, Van Aelst A, Van As H, Versprille AI, Lettinga G (2001) Cluster structure of anaerobic aggregates of an expanded granular sludge bed reactor. Appl Environ Microbiol 67:3683–3692

  25. Guibaud G, Tixier N, Bouju A, Baudu M (2003) Relation between extracellular polymers' composition and its ability to complex Cd, Cu and Pb. Chemosphere 52:1701–1710

  26. Guibaud G, van Hullebusch E, Bordas F (2006) Lead and cadmium biosorption by extracellular polymeric substances (EPS) extracted from activated sludges: pH-sorption edge tests and mathematical equilibrium modelling. Chemosphere 64:1955–1962

  27. Guibaud G, Bordas F, Saaid A, d'Abzac P, Van Hullebusch E (2008) Effect of pH on cadmium and lead binding by extracellular polymeric substances (EPS) extracted from environmental bacterial strains. Colloids Surf B: Biointerfaces 63:48–54

  28. Guibaud G, van Hullebusch E, Bordas F, d’Abzac P, Joussein E (2009) Sorption of Cd(II) and Pb(II) by exopolymeric substances (EPS) extracted from activated sludges and pure bacterial strains: modeling of the metal/ligand ratio effect and role of the mineral fraction. Bioresource Technology 100:2959–2968

  29. Harmita H, Karthikeyan KG, Pan X (2009) Copper and cadmium sorption onto kraft and organosolv lignins. Bioresour Technol 100:6183–6191

  30. Hawari AH, Mulligan CN (2006) Biosorption of lead(II), cadmium(II), copper(II) and nickel(II) by anaerobic granular biomass. Bioresour Technol 97:692–700

  31. Hulshoff Pol LW, De Castro Lopes SI, Lettinga G, Lens PNL (2004) Anaerobic sludge granulation. Water Res 38:1376–1389

  32. Li J, Liu X, Liu Y, Ramsay J, Yao C, Dai R (2011) The effect of continuous exposure of copper on the properties and extracellular polymeric substances (EPS) of bulking activated sludge. Environ Sci Pollut Res 18:1567–1573

  33. Joshi PM, Juwarkar AA (2009) In vivo studies to elucidate the role of extracellular polymeric substances from Azotobacter in immobilization of heavy metals. Environ Sci Technol 43:5884–5889

  34. Kassam ZA, Yerushalmi L, Guiot SR (2003) A market study on the anaerobic wastewater treatment systems. Water Air Soil Pollut 143:179–192

  35. Lamelas C, Benedetti M, Wilkinson KJ, Slaveykova VI (2006) Characterization of H+ and Cd2+ binding properties of the bacterial exopolysaccharides. Chemosphere 65:1362–1370

  36. Lartiges BS, Deneux-Mustin S, Villemin G, Mustin C, Barres O, Chamerois M, Gerard B, Babut M (2001) Composition, structure and size distribution of suspended particulates from the Rhine River. Water Res 35:808–816

  37. Laurent J, Casellas M, Dagot C (2009) Heavy metals uptake by sonicated activated sludge: relation with floc surface properties. J Hazard Mater 162:652–660

  38. Liu H, Fang HHP (2002) Characterization of electrostatic binding sites of extracellular polymers by linear programming analysis of titration data. Biotechnol Bioeng 80:806–811

  39. Liu Y, Fang HHP (2003) Influences of extracellular polymeric substances (EPS) on flocculation, settling, and dewatering of activated sludge. Crit Rev Environ Sci Technol 33:237–273

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

  41. Lores EM, Pennock JR (1998) The effect of salinity on binding of Cd, Cr, Cu and Zn to dissolved organic matter. Chemosphere 37:861–874

  42. Lowry O, Rosebrough N, Fan A, Randall R (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

  43. Mañas A, Spérandio M, Decker F, Biscans B (2012) Location and chemical composition of microbially induced phosphorus precipitates in anaerobic and aerobic granular sludge. Environ Technol 33:2195–2209

  44. Matheickal JT, Yu Q, Woodburn GM (1999) Biosorption of cadmium(II) from aqueous solutions by pre-treated biomass of marine alga Durvillaea potatorum. Water Res 33:335–342

  45. Mikutta R, Baumgärtner A, Schippers A, Haumaier L, Guggenberger G (2012) Extracellular polymeric substances from Bacillus subtilis associated with minerals modify the extent and rate of heavy metal sorption. Environ Sci Technol 46:3866–3873

  46. Metreveli G, Abbt-Braun G, Frimmel FH (2009): Influence of NOM on the Mobility of Metal(loid)s in Water-Saturated Porous Media. Aquatic Geochemistry, 1–16

  47. Nakanishi K, Solomon PH (eds) (1977) Infrared absorption spectroscopy. Holden-Day, Inc., San Francisco

  48. Oliveira ADS, Bodo A, Beltramini Trevilato TM, Magosso Takayanagui AM, Domingo JL, Segura-MuÃoz SI (2007) Heavy metals in untreated/treated urban effluent and sludge from a biological wastewater treatment plant. Environ Sci Pollut Res 14:483–489

  49. Omoike A, Chorover J (2006) Adsorption to goethite of extracellular polymeric substances from Bacillus subtilis. Geochim Cosmochim Acta 70:827–838

  50. Ozturk S, Aslim B (2008) Relationship between chromium(VI) resistance and extracellular polymeric substances (EPS) concentration by some cyanobacterial isolates. Environ Sci Pollut Res 15:478–480

  51. Pal A, Paul AK (2008) Microbial extracellular polymeric substances: central elements in heavy metal bioremediation. Indian J Microbiol 48:49–64

  52. Pambrun V, Marquot A, Racault Y (2008) Characterization of the toxic effects of cadmium and 3.5-dichlorophenol on nitrifying activity and mortality in biologically activated sludge systems—effect of low temperature. Environ Sci Pollut Res 15:592–599

  53. Peña J, Bargar JR, Sposito G (2011) Role of bacterial biomass in the sorption of Ni by biomass–birnessite assemblages. Environ Sci Technol 45:7338–7344

  54. Sag Y, Kutsal T (1995) Biosorption of heavy metals by Zoogloea ramigera: use of adsorption isotherms and a comparison of biosorption characteristics. Chem Eng J Biochem Eng J 60:181–188

  55. Santín C, Yamashita Y, Otero XL, Álvarez MÁ, Jaffé R (2009) Characterizing humic substances from estuarine soils and sediments by excitation-emission matrix spectroscopy and parallel factor analysis. Biogeochemistry 96:131–147

  56. Shirvani M, Kalbasi M, Shariatmadari H, Nourbakhsh F, Najafi B (2006) Sorption–desorption of cadmium in aqueous palygorskite, sepiolite, and calcite suspensions: isotherm hysteresis. Chemosphere 65:2178–2184

  57. Sipos P, Németh T, Kis VK, Mohai I (2009) Association of individual soil mineral constituents and heavy metals as studied by sorption experiments and analytical electron microscopy analyses. J Hazard Mater 168:1512–1520

  58. Stone EA, Hedman CJ, Sheesley RJ, Shafer MM, Schauer JJ (2009) Investigating the chemical nature of humic-like substances (HULIS) in North American atmospheric aerosols by liquid chromatography tandem mass spectrometry. Atmosp Environ 43:4205–4213

  59. Sun X-F, Wang S-G, Zhang X-M, Paul Chen J, Li X-M, Gao B-Y, Ma Y (2009) Spectroscopic study of Zn2+ and Co2+ binding to extracellular polymeric substances (EPS) from aerobic granules. J Colloid Interface Sci 335:11–17

  60. Tipping E (1993) Modelling ion binding by humic acids. Colloids Surf, A Physicochem Eng Asp 73:117–131

  61. Tourney J, Ngwenya BT (2009) Bacterial extracellular polymeric substances (EPS) mediate CaCO3 morphology and polymorphism. Chem Geol 262:138–146

  62. Turner BF, Fein JB (2006) Protofit: a program for determining surface protonation constants from titration data. Comput Geosci 32:1344–1356

  63. Ueshima M, Ginn BR, Haack EA, Szymanowski JES, Fein JB (2008) Cd adsorption onto Pseudomonas putida in the presence and absence of extracellular polymeric substances. Geochim Cosmochim Acta 72:5885–5895

  64. Van Langerak EPA, Gonzalez-Gil G, Van Aelst A, Van Lier JB, Hamelers HVM, Lettinga G (1998) Effects of high calcium concentrations on the development of methanogenic sludge in upflow anaerobic sludge bed (UASB) reactors. Water Res 32:1255–1263

  65. Vinod VTP, Sashidhar RB, Sreedhar B, Rama Rao B, Nageswara Rao T, Abraham JT (2009) Interaction of Pb2+ and Cd2+ with gum kondagogu (Cochlospermum gossypium): a natural carbohydrate polymer with biosorbent properties. Carbohydr Polym 78:894–901

  66. Wingender J, Neu TR, Flemming HC (1999) Microbial extracellular polymeric substances: characterisation, structure and function. Springer, New York

  67. Yuncu B, Sanin FD, Yetis U (2006) An investigation of heavy metal biosorption in relation to C/N ratio of activated sludge. J Hazard Mater 137:990–997

  68. Zhu R, Yu R, Yao J, Mao D, Xing C, Wang D (2008) Removal of Cd2+ from aqueous solutions by hydroxyapatite. Catal Today 139:94–99

  69. Zhuang J, Yu GR (2002) Effects of surface coatings on electrochemical properties and contaminant sorption of clay minerals. Chemosphere 49:619–628

Download references

Acknowledgment

This study was supported by a research grant provided by the Conseil Regional du Limousin.

Author information

Correspondence to Gilles Guibaud.

Additional information

Submitted to Environmental Science and Pollution Research, ESPR-D-12-00929

Responsible editor: Philippe Garrigues

Rights and permissions

Reprints and Permissions

About this article

Cite this article

d’Abzac, P., Bordas, F., Joussein, E. et al. Metal binding properties of extracellular polymeric substances extracted from anaerobic granular sludges. Environ Sci Pollut Res 20, 4509–4519 (2013). https://doi.org/10.1007/s11356-012-1401-3

Download citation

Keywords

  • Anaerobic granular sludge
  • Extracellular polymeric substances (EPS)
  • EPS characterization
  • Acidic constant
  • Metal sorption