Environmental Science and Pollution Research

, Volume 26, Issue 4, pp 3741–3750 | Cite as

Effects of different types of extracellular polysaccharides isolated from cyanobacterial blooms on the colony formation of unicellular Microcystis aeruginosa

  • Ken Omori
  • Tania Datta
  • Yoshimasa AmanoEmail author
  • Motoi Machida
Research Article


In this study, two types of extracellular polysaccharides (EPS), namely, mixed EPS (MX-EPS) and tightly bound EPS (TB-EPS), were extracted from cyanobacterial blooms using different methods. To evaluate their compositional differences, elemental composition, FTIR, and TG/DTA profile were measured for both EPS samples. Following that, unicellular Microcystis aeruginosa was cultured in a medium containing EPS, Ca2+ ion, and Mg2+ ion, and the effect of each type of EPS on the colony formation of M. aeruginosa was examined. Results showed that TB-EPS had more carboxy groups than MX-EPS, and that the TB-EPS medium contained Ca2+ and Mg2+ ions. These cations were not detected in the MX-EPS medium. During the colony formation experiment, colonies were observed when Ca2+ and Mg2+ ions were present at 250 mg/L concentration each. In addition, colony density increased when TB-EPS was added, compared to that of MX-EPS. Colonies were also observed in the medium containing only TB-EPS (100 mg/L), indicating that M. aeruginosa can form colonies using Ca2+ ion present in TB-EPS. During the MX-EPS extraction, Ca2+ ion chelated with EDTA was removed during ethanol precipitation. Therefore, the extraction protocol followed for TB-EPS was better than that of MX-EPS for maintaining Ca2+ ions, and thereby maintaining an EPS composition that enables for colony formation.


Cyanobacterial blooms Microcystis aeruginosa Extracellular polysaccharides (EPS) Cation Extraction Colony formation 



The authors would like to extend deep gratitude to Prof. Dr. Fumio Imazeki, Safety and Health Organization, Chiba University, for the encouragement of this study.

Funding information

This work was partially supported by JFE 21st Century Foundation and by the Japan Society for the Promotion of Science (JSPS) under a Grant-in-Aid for Scientific Research (C) (No. 18 K04404).


  1. Amemiya Y, Nakayama O (1984) The chemical composition and metal adsorption capacity of the sheath materials isolated from Microcystis, Cyanobacteria. Jpn J Limnol 45:187–193 (in Japanese)CrossRefGoogle Scholar
  2. Bi X, Dai W, Zhang S, Xing K, Zhang X (2015) Accumulation and distribution characteristics of heavy metals in different size Microcystis colonies from natural waters. Fresenius Environ Bull 24:773–779Google Scholar
  3. Dubois M, Gilles KA, Hamilton JK, Rebers PT, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356Google Scholar
  4. Fattom A, Shilo M (1984) Hydrophobicity as an adhesion mechanism of benthic cyanobacteria. Appl Environ Microbiol 47:135–143Google Scholar
  5. Figueiredo JL, Pereira MFR, Freitas MMA, Orfao JJM (1999) Modification of the surface chemistry of activated carbons. Carbon 37:1379–1389CrossRefGoogle Scholar
  6. Forni C, Telo’ FR, Caiola MG (1997) Comparative analysis of the polysaccharides produced by different species of Microcystis (Chroococcales, Cyanophyta). Phycologia 36:181–185CrossRefGoogle Scholar
  7. Graham JL, Loftin KA, Meyer MT, Ziegler AC (2010) Cyanotoxin mixtures and taste-and-odor compounds in cyanobacterial blooms from the Midwestern United States. Environ Sci Technol 44:7361–7368CrossRefGoogle Scholar
  8. Guillard RR, Lorenzen CJ (1972) Yellow-green algae with chlorophyllide C. J Phycol 8:10–14Google Scholar
  9. Hadjoudja S, Deluchat V, Baudu M (2010) Cell surface characterisation of Microcystis aeruginosa and Chlorella vulgaris. J Colloid Interface Sci 342:293–299CrossRefGoogle Scholar
  10. Hassler CS, Alasonati E, Nichols CM, Slaveykova VI (2011) Exopolysaccharides produced by bacteria isolated from the pelagic Southern Ocean - role in Fe binding, chemical reactivity, and bioavailability. Mar Chem 123:88–98CrossRefGoogle Scholar
  11. Hou J, Yang Y, Wang P, Wang C, Miao L, Wang X, Lv B, You G, Liu Z (2017) Effects of CeO2, CuO, and ZnO nanoparticles on physiological features of Microcystis aeruginosa and the production and composition of extracellular polymeric substances. Environ Sci Pollut Res 24:226–235CrossRefGoogle Scholar
  12. Jia YF, Thomas KM (2000) Adsorption of cadmium ions on oxygen surface sites in activated carbon. Langmuir 16:1114–1122CrossRefGoogle Scholar
  13. Li P, Cai Y, Shi L, Geng L, Xing P, Yu Y, Kong F, Wang Y (2009) Microbial degradation and preliminary chemical characterization of Microcystis exopolysaccharides from a cyanobacterial water bloom of Lake Taihu. Int Rev Hydrobiol 94:645–655CrossRefGoogle Scholar
  14. Li M, Zhu W, Gao L, Lu L (2013) Changes in extracellular polysaccharide content and morphology of Microcystis aeruginosa at different specific growth rates. J Appl Phycol 25:1023–1030CrossRefGoogle Scholar
  15. Li M, Zhu W, Sun Q (2014) Solubilisation of mucilage induces changes in Microcystis colonial morphology. N Z J Mar Freshw Res 48:38–47CrossRefGoogle Scholar
  16. Liu Y, Wang W, Geng L, Chen Y, Yang Z (2011) Polysaccharide content and morphology of Microcystis aeruginosa in response to changes in metabolic carbon flux. Fresenius Environ Bull 20:1046–1050Google Scholar
  17. Medrano EA, Uittenbogaard RE, Pires LD, Van De Wiel BJH, Clercx HJH (2013) Coupling hydrodynamics and buoyancy regulation in Microcystis aeruginosa for its vertical distribution in lakes. Ecol Model 248:41–56CrossRefGoogle Scholar
  18. Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta 27:31–36Google Scholar
  19. Nishikawa S, Kuriyama M (1974) Nucleic acid as a component of mucilage in activated sludge. J Ferment Technol 52:335–338 (in Japanese)Google Scholar
  20. Omori K, Sato M, Amano Y, Machida M (2018) Induction of colony formation of Microcystis aeruginosa by controlling extracellular polysaccharides and metal cation concentrations. J Chem Eng Jpn 51:289–297CrossRefGoogle Scholar
  21. Otsuka S, Suda S, Li R, Matsumoto S, Watanabe MM (2000) Morphological variability of colonies of Microcystis morphospecies in culture. J Gen Appl Microbiol 46:39–50CrossRefGoogle Scholar
  22. Paerl HW, Huisman J (2008) Blooms like it hot. Science 320:57–58CrossRefGoogle Scholar
  23. Qu F, Liang H, He J, Ma J, Wang Z, Yu H, Li G (2012) Characterization of dissolved extracellular organic matter (dEOM) and bound extracellular organic matter (bEOM) of Microcystis aeruginosa and their impacts on UF membrane fouling. Water Res 46:2881–2890CrossRefGoogle Scholar
  24. Qu F, Du X, Liu B, He J, Ren N, Li G, Liang H (2015) Control of ultrafiltration membrane fouling caused by Microcystis cells with permanganate preoxidation: significance of in situ formed manganese dioxide. Chem Eng J 279:56–65CrossRefGoogle Scholar
  25. Reynolds CS (1987) Cyanobacterial water blooms. In: Callow J (ed) Advances in botanical research. Academic Press, London, pp 67–143Google Scholar
  26. Reynolds CS (2007) Variability in the provision and function of mucilage in phytoplankton: facultative responses to the environment. Hydrobiologia 578:37–45CrossRefGoogle Scholar
  27. Sato M, Amano Y, Machida M, Imazeki F (2017a) Colony formation of highly dispersed Microcystis aeruginosa by controlling extracellular polysaccharides and calcium ion concentrations in aquatic solution. Limnology 18:111–119CrossRefGoogle Scholar
  28. Sato M, Omori K, Datta T, Amano Y, Machida M (2017b) Influence of extracellular polysaccharides and calcium ion on colony formation of unicellular Microcystis aeruginosa. Environ Eng Sci 34:149–157CrossRefGoogle Scholar
  29. Sheng GP, Yu HQ, Yu Z (2005) Extraction of extracellular polymeric substances from the photosynthetic bacterium Rhodopseudomonas acidophila. Appl Microbiol Biotechnol 67:125–130CrossRefGoogle Scholar
  30. Sheng GP, Yu HQ, Li XY (2010) Extracellular polymeric substances (EPS) of microbial aggregates in biological wastewater treatment systems: a review. Biotechnol Adv 28:882–894CrossRefGoogle Scholar
  31. Shi JQ, Wu ZX, Song LR (2013) Physiological and molecular responses to calcium supplementation in Microcystis aeruginosa (Cyanobacteria). N Z J Mar Freshw Res 47:51–61CrossRefGoogle Scholar
  32. Toyoshima C (2008) Structural aspects of ion pumping by Ca2+ -ATPase of sarcoplasmic reticulum. Arch Biochem Biophys 476:3–11CrossRefGoogle Scholar
  33. Walsby AE, Hayes PK, Boje R (1995) The gas vesicles, buoyancy and vertical distribution of cyanobacteria in the Baltic Sea. Eur J Phycol 30:87–94CrossRefGoogle Scholar
  34. Wang YW, Zhao J, Li JH, Li SS, Zhang LH, Wu M (2011) Effects of calcium levels on colonial aggregation and buoyancy of Microcystis aeruginosa. Curr Microbiol 62:679–683CrossRefGoogle Scholar
  35. Wang LL, Wang LF, Ren XM, Ye XD, Li WW, Yuan SJ, Sun M, Sheng GP, Yu HQ, Wang XK (2012) pH dependence of structure and surface properties of microbial EPS. Environ Sci Technol 46:737–744CrossRefGoogle Scholar
  36. Xiao M, Willis A, Burford MA, Li M (2017) Review: a meta-analysis comparing cell-division and cell-adhesion in Microcystis colony formation. Harmful Algae 67:85–91CrossRefGoogle Scholar
  37. Xiao M, Li M, Reynolds CS (2018) Colony formation in the cyanobacterium Microcystis. Biol Rev 93:1399–1420CrossRefGoogle Scholar
  38. Xu H, Yu G, Jiang H (2013) Investigation on extracellular polymeric substances from mucilaginous cyanobacterial blooms in eutrophic freshwater lakes. Chemosphere 93:75–81CrossRefGoogle Scholar
  39. Yamamoto Y, Nakahara H (2009) Seasonal variations in the morphology of bloom-forming cyanobacteria in a eutrophic pond. Limnology 10:185–193CrossRefGoogle Scholar
  40. Yang Z, Kong F (2013) Abiotic factors in colony formation: effects of nutrition and light on extracellular polysaccharide production and cell aggregates of Microcystis aeruginosa. Chin J Oceanol Limnol 31:796–802CrossRefGoogle Scholar
  41. Yang Z, Kong F, Shi X, Zhang M, Xing P, Cao H (2008) Changes in the morphology and polysaccharide content of Microcystis aeruginosa (Cyanobacteria) during flagellate grazing. J Phycol 44:716–720CrossRefGoogle Scholar
  42. Zhang K, Lin TF, Zhang T, Li C, Gao N (2013) Characterization of typical taste and odor compounds formed by Microcystis aeruginosa. J Environ Sci 25:1539–1548CrossRefGoogle Scholar
  43. Zhao L, Lu L, Li M, Xu Z, Zhu W (2011) Effects of Ca and Mg levels on colony formation and EPS content of cultured M. aeruginosa. Procedia Environ Sci 10:1452–1458CrossRefGoogle Scholar
  44. Zhu W, Dai X, Li M (2014a) Relationship between extracellular polysaccharide (EPS) content and colony size of Microcystis is colonial morphology dependent. Biochem Syst Ecol 55:346–350CrossRefGoogle Scholar
  45. Zhu W, Li M, Luo Y, Dai X, Guo L, Xiao M, Huang J, Tan X (2014b) Vertical distribution of Microcystis colony size in Lake Taihu: its role in algal blooms. J Gt Lakes Res 40:949–955CrossRefGoogle Scholar
  46. Zou H, Pan G, Chen H, Yuan X (2006) Removal of cyanobacterial blooms in Taihu Lake using local soils II. Effective removal of Microcystis aeruginosa using local soils and sediments modified by chitosan. Environ Pollut 141:201–205CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Ken Omori
    • 1
  • Tania Datta
    • 2
  • Yoshimasa Amano
    • 1
    • 3
    Email author
  • Motoi Machida
    • 1
    • 3
  1. 1.Graduate School of EngineeringChiba UniversityChibaJapan
  2. 2.Center for the Management, Utilization and Protection of Water ResourcesTennessee Technological UniversityCookevilleUSA
  3. 3.Safety and Health OrganizationChiba UniversityChibaJapan

Personalised recommendations