Skip to main content
SpringerLink
Log in

Extraction and characterization of extracellular polymeric substances from a mixed fungal culture during the adaptation process with waste printed circuit boards

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Extracellular polymeric substances (EPSs) extracted from fungal mycelium by four chemical methods (NaOH, H2SO4, formaldehyde-NaOH, glutaraldehyde-NaOH), three physical methods (heating, ultrasound, vibration), and a control method (centrifugation alone) were investigated. Results indicated formaldehyde-NaOH outperformed other methods with 186.6 ± 8.0 mg/g of polysaccharides and 23.2 ± 4.6 mg/g of protein extracted and ensured little contamination by intracellular substances. Thereafter, this method was applied in extracting EPS from a mixed fungal culture in the adaptation process with 0.5% (w/v) waste printed circuit boards (PCBs). With the four adaptation stages continuing, the culture tended to become more sensitive to respond to the external toxic environment characterized by secreting EPS more easily and quickly. The maximum amount of polysaccharides and protein could be achieved in only 3 days both at the 3rd and 4th adaptation stage. Three-dimensional excitation-emission matrix fluorescence spectrum indicated the peaks obtained for EPS were mainly associated to soluble microbial by-product-like and aromatic protein-like compounds. Transmission electron microscopic observation illustrated that although metal ions penetrated into hypha cells, parts of them could be absorbed by EPS, implying that EPS secretion may be a primary protective strategy adopted by the culture.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

References

  • Adav SS, Lee DJ (2008) Extraction of extracellular polymeric substances from aerobic granule with compact interior structure. J Hazard Mater 154:1120–1126

    Article  CAS  Google Scholar 

  • Amiri F, Yaghmaei S, Mousavi SM, Sheibani S (2011) Recovery of metals from spent refinery hydrocracking catalyst using adapted Aspergillus niger. Hydrometallurgy 109:65–71

    Article  CAS  Google Scholar 

  • Bai J, Chao Y, Chen Y, Wang S, Qiu R (2017) Immobilization of Cu by Bacillus subtilis DBM and the role of extracellular polymeric substances. Water Air Soil Pollut 228:86

    Article  CAS  Google Scholar 

  • Baker A (2001) Fluorescence excitation-emission matrix characterization of some sewage-impacted rivers. Environ Sci Technol 35:948–953

    Article  CAS  Google Scholar 

  • Beech WB, Sunner J (2004) Biocorrosion: towards understanding interactions between biofilms and metals. Curr Opin Biotechnol 15:181–186

    Article  CAS  Google Scholar 

  • Bellion M, Courbot M, Jacob C, Blaudez D, Chalot M (2006) Extracellular and cellular mechanisms sustaining metal tolerance in ectomycorrhizal fungi. FEMS Microbiol Lett 254:173–181

    Article  CAS  Google Scholar 

  • Burgstaller W, Schinner F (1993) Leaching of metals with fungi. J Biotechnol 27:91–116

    Article  CAS  Google Scholar 

  • Chen W, Westerhoff P, Leenheer JA, Booksh K (2003) Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter. Environ Sci Technol 37:5701–5710

    Article  CAS  Google Scholar 

  • Chen X, Shi J, Chen Y, Xu X, Xu S, Wang Y (2006) Tolerance and biosorption of copper and zinc by Pseudomonas putida CZ1 isolated from metal-polluted soil. Can J Microbiol 52:308–316

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Domínguez L, Rodríguez M, Prats D (2010) Effect of different extraction methods on bound EPS from MBR sludges. Part I: influence of extraction methods over three-dimensional EEM fluorescence spectroscopy fingerprint. Desalination 261:19–26

    Article  CAS  Google Scholar 

  • Gong AS, Bolster CH, Benavides M, Walker SL (2009) Extraction and analysis of extracellular polymeric substances: comparison of methods and extracellular polymeric substance levels in Salmonella pullorum SA 1685. Environ Eng Sci 26:1523–1532

    Article  CAS  Google Scholar 

  • Jablonski JE, Fu T-J, Jackson LS, Gendel SM (2010) Determination of protein levels in soy and peanut oils by colorimetric assay and ELISA. J AOAC Int 93:213–220

    CAS  Google Scholar 

  • Kelleher BP, Simpson AJ (2006) Humic substances in soils: are they really chemically distinct? Environ Sci Technol 40:4605–4611

    Article  CAS  Google Scholar 

  • Kinzler K, Gehrke T, Telegdi J, Sand W (2003) Bioleaching - a result of interfacial processes caused by extracellular polymeric substances (EPS). Hydrometallurgy 71:83–88

    Article  CAS  Google Scholar 

  • Kleber M, Johnson MG (2010) Chapter 3 – advances in understanding the molecular structure of soil organic matter : implications for interactions in the environment. Adv Agron 106:77–142

    Article  CAS  Google Scholar 

  • Liu H, Fang HHP (2002) Extraction of extracellular polymeric substances (EPS) of sludges. J Biotechnol 95:249–256

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Lu Q, Chang M, Yu Z, Zhou S (2015) The effects of three commonly used extraction methods on the redox properties of extracellular polymeric substances from activated sludge. Environ Technol 36:2884–2891

    Article  CAS  Google Scholar 

  • McSwain BS, Irvine RL, Hausner M, Wilderer PA (2005) Composition and distribution of extracellular polymeric substances in aerobic flocs and granular sludge. Appl Environ Microbiol 71:1051–1057

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Peng TJ, Zhou D, Liu YN, Yu RL, Qiu GZ, Zeng WM (2019): Effects of pH value on the expression of key iron/sulfur oxidation genes during bioleaching of chalcopyrite on thermophilic condition. https://doi.org/10.1007/s13213-019-01453-y

  • Santamaria M, Diaz-Marrero AR, Hernandez J, Gutierrez-Navarro AM, Corzo J (2003) Effect of thorium on the growth and capsule morphology of Bradyrhizobium. Environ Microbiol 5:916–924

    Article  CAS  Google Scholar 

  • Sheng G-P, Yu H-Q, Li X-Y (2010) Extracellular polymeric substances (EPS) of microbial aggregates in biological wastewater treatment systems: a review. Biotechnol Adv 28:882–894

    Article  CAS  Google Scholar 

  • Sheng GP, Yu HQ, Yu Z (2005) Extraction of extracellular polymeric substances from the photosynthetic bacterium Rhodopseudomonas acidophila. Appl Microbiol Biotechnol 67:125–130

    Article  CAS  Google Scholar 

  • Sobeck DC, Higgins MJ (2002) Examination of three theories for mechanisms of cation-induced bioflocculation. Water Res 36:527–538

    Article  CAS  Google Scholar 

  • Tan SN, Chen M (2012) Early stage adsorption behaviour of Acidithiobacillus ferrooxidans on minerals I: an experimental approach. Hydrometallurgy 119:87–94

    Article  CAS  Google Scholar 

  • Urrutia MM, Beveridge TJ (1993) Remobilization of heavy metals retained as oxyhydroxides or silicates by Bacillus subtilis cells. Appl Environ Microbiol 59:4323–4329

    CAS  Google Scholar 

  • Wingender J, Neu TR, Flemming HC (1999) Microbial extracellular polymeric substances. Springer Berlin Heidelberg, 45-53

  • Wu XL, Wu XY, Shen L, Li JK, Yu RL, Liu YD, Qiu GZ, Zeng WM (2019): Whole genome sequencing and comparative genomics analyses of Pandoraea sp. XY-2, a new species capable of biodegrade tetracycline. Front Microbiol 10

  • Xia M, Wang Y, Peng T, Shen L, Yu R, Liu Y, Chen M, Li J, Wu X, Zeng W (2017) Recycling of metals from pretreated waste printed circuit boards effectively in stirred tank reactor by a moderately thermophilic culture. J Biosci Bioeng 123:714–721

    Article  CAS  Google Scholar 

  • Xia M, Bao P, Liu A, Wang M, Shen L, Yu R, Liu Y, Chen M, Li J, Wu X (2018) Bioleaching of low-grade waste printed circuit boards by mixed fungal culture and its community structure analysis. Resour Conserv Recycl 136:267–275

    Article  Google Scholar 

  • Yu R, Hou C, Liu A, Peng T, Xia M, Wu X, Shen L, Liu Y, Li J, Yang F, Qiu G, Chen M, Zeng W (2018) Extracellular DNA enhances the adsorption of Sulfobacillus thermosulfidooxidans strain ST on chalcopyrite surface. Hydrometallurgy 176:97–103

    Article  CAS  Google Scholar 

  • Yu RL, Liu A, Liu Y, Yu Z, Peng T, Wu X, Shen L, Liu Y, Li J, Liu X, Qiu G, Chen M, Zeng W (2017) Evolution of Sulfobacillus thermosulfidooxidans secreting alginate during bioleaching of chalcopyrite concentrate. J Appl Microbiol 122:1586–1594

    Article  CAS  Google Scholar 

  • Zeng W, Qiu G, Zhou H, Liu X, Chen M, Chao W, Zhang C, Peng J (2010) Characterization of extracellular polymeric substances extracted during the bioleaching of chalcopyrite concentrate. Hydrometallurgy 100:177–180

    Article  CAS  Google Scholar 

  • Zhang L, Ren H, Ding L (2012) Comparison of extracellular polymeric substances (EPS) extraction from two different activated sludges. Water Sci Technol 66:1558–1564

    Article  CAS  Google Scholar 

  • Zhang X, Bishop PL (2003) Biodegradability of biofilm extracellular polymeric substances. Chemosphere 50:63–69

    Article  CAS  Google Scholar 

  • Zheng Y, Zhao X, Jia K, Li J, Wang L, Wang R, Lu X, Zhang D (2018) bifA regulates biofilm development of Pseudomonas putidaMnB1 as a primary response to H2O2and Mn2+. Front Microbiol 9:1490

    Article  Google Scholar 

  • Zhu L, Qi H, Lv M, Kong Y, Yu Y, Xu X (2012) Component analysis of extracellular polymeric substances (EPS) during aerobic sludge granulation using FTIR and 3D-EEM technologies. Bioresour Technol 124:455–459

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the National Natural Science Foundation of China (No. 31470230, 51320105006, 51604308), the Youth Talent Foundation of Hunan Province of China (No.2017RS3003), Natural Science Foundation of Hunan Province of China (No.2018JJ2486), and Key Research and Development Projects in Hunan Province (2018WK2012).

Author information

Authors and Affiliations

  1. School of Minerals Processing and Bioengineering, Central South University, Changsha, 410083, China

    Mingchen Xia, Peng Bao, Shishi Zhang, Ajuan Liu, Li Shen, Runlan Yu, Yuandong Liu, Jiaokun Li, Xueling Wu, Guanzhou Qiu & Weimin Zeng

  2. Key Laboratory of Biometallurgy, Ministry of Education, Changsha, 410083, China

    Li Shen, Runlan Yu, Yuandong Liu, Jiaokun Li, Xueling Wu, Guanzhou Qiu & Weimin Zeng

  3. CSIRO Process Science and Engineering, Clayton, Victoria, 3168, Australia

    Miao Chen & Weimin Zeng

  4. Centre for Advanced Materials and Industrial Chemistry, RMIT University, Melbourne, 3000, Australia

    Miao Chen

Authors
  1. Mingchen Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peng Bao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shishi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ajuan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Li Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Runlan Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yuandong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Miao Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jiaokun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Xueling Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Guanzhou Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Weimin Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Weimin Zeng.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Additional information

Responsible editor: Philippe Garrigues

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xia, M., Bao, P., Zhang, S. et al. Extraction and characterization of extracellular polymeric substances from a mixed fungal culture during the adaptation process with waste printed circuit boards. Environ Sci Pollut Res 26, 22137–22146 (2019). https://doi.org/10.1007/s11356-019-05234-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-019-05234-7

Keywords

Search

Navigation