Applied Microbiology and Biotechnology

, Volume 102, Issue 12, pp 5343–5353 | Cite as

The attachment potential and N-acyl-homoserine lactone-based quorum sensing in aerobic granular sludge and algal-bacterial granular sludge

  • Bing Zhang
  • Piet N. L. Lens
  • Wenxin Shi
  • Ruijun Zhang
  • Zhiqiang Zhang
  • Yuan Guo
  • Xian Bao
  • Fuyi Cui
Environmental biotechnology


Bacteria and algae often coexist in the aerobic granular sludge (AGS) system in a photo-bioreactor, forming algal-bacterial granular sludge. In this study, the physicochemical characteristics and microbial attachment potential of the AGS and algal-bacterial granular sludge were comparatively analyzed. Results clearly showed that the larger and denser algal-bacterial granular sludge had stronger attachment potential compared to the AGS (as the control). A bioassay with Agrobacterium tumefaciens KYC55 indicated that N-acyl-homoserine lactones (AHLs) existed in both sludge types, but further investigations revealed that the relative AHL content of the algal-bacterial granular sludge obviously increased and slightly decreased during phases II and III, respectively, but was consistently higher than the AGS. Based on the EPS measurements and 3D-excitation-emission matrix (3D-EEM) fluorescence spectra analysis, the enhancement of AHL-based QS favored the hydrophobic protein production of algal-bacterial granular sludge, contributing to a good development of the granular sludge. In addition, it was also found that inhibition of AHLs resulted in the reduction of the protein content and attachment potential in algal-bacterial granular sludge, which was unfavorable to the structural stability of the granules. High-throughput sequencing analysis showed that the microbial community of AGS was different from the algal-bacterial granular sludge; specifically, algal-bacterial granulation facilitated the abundance of AHLs and EPS producers, such as the genera Acinetobacter, Chryseobacterium, and Flavobacterium.


Algal-bacterial granules Quorum sensing (QS) N-acyl-homoserine lactone (AHL) Attachment potential Extracellular polymeric substances (EPS) 



The authors would like to show our sincerely gratitude for Prof. Hui Wang in Nanjing Agricultural University for providing Agrobacterium tumefaciens KYC55 (pJ372) (pJ384) (pJ410). This work was financed by the National Natural Science Foundation of China (51778172), State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology (2016DX11), and the Nanqi Ren Studio, Academy of Environment & Ecology, Harbin Institute of Technology.

Compliance with ethical standards

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

253_2018_9002_MOESM1_ESM.pdf (415 kb)
ESM 1 (PDF 414 kb)


  1. Adav SS, Duujong L, Lai JY (2010) Potential cause of aerobic granular sludge breakdown at high organic loading rates. Appl Microbiol Biotechnol 85(5):1601–1610CrossRefPubMedGoogle Scholar
  2. Amin SA, Hmelo LR, van Tol HM, Durham BP, Carlson LT, Heal KR, Morales RL, Berthiaume CT, Parker MS, Djunaedi B (2015) Interaction and signalling between a cosmopolitan phytoplankton and associated bacteria. Nature 522:98–101CrossRefPubMedGoogle Scholar
  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(s 1-2):248–254CrossRefPubMedGoogle Scholar
  4. Byunghyuk K, Ramanan R, Daehyun C, Heemock O, Heesik K (2014) Role of Rhizobium, a plant growth promoting bacterium, in enhancing algal biomass through mutualistic interaction. Biomass Bioenergy 69(3):95–105Google Scholar
  5. Chabalina LD, Pastor MR, Rico DP (2013) Characterization of soluble and bound EPS obtained from 2 submerged membrane bioreactors by 3D-EEM and HPSEC. Talanta 115:706–712CrossRefGoogle Scholar
  6. Cho DH, Ramanan R, Kim BH, Lee J, Kim S, Chan Y, Choi GG, Oh HM, Kim HS (2013) Novel approach for the development of axenic microalgal cultures from environmental samples. J Phycol 49(4):802–810CrossRefPubMedGoogle Scholar
  7. Cjs B, Subramanian TA, Green DH (2011) The toxic dinoflagellate gymnodinium catenatum (dinophyceae) requires marine bacteria for growth. J Phycol 47(5):1009–1022CrossRefGoogle Scholar
  8. Cooper MB, Smith AG (2015) Exploring mutualistic interactions between microalgae and bacteria in the omics age. Curr Opin Plant Biol 26:147–153CrossRefPubMedGoogle Scholar
  9. Dahalan FA, Abdullah N, Yuzir A, Olsson G, Salmiati, Hamdzah M, Din MFM, Ahmad SA, Khalil KA, Anuar AN, Noor ZZ, Ujang Z (2015) A proposed aerobic granules size development scheme for aerobic granulation process. Bioresour Technol 181:291–296CrossRefPubMedGoogle Scholar
  10. de Kreuk MK, Heijnen JJ, van Loosdrecht MC (2005) Simultaneous COD, nitrogen, and phosphate removal by aerobic granular sludge. Biotechnol Bioeng 90(6):761–769CrossRefPubMedGoogle Scholar
  11. de la Fuente-Núñez C, Reffuveille F, Fernández L, Hancock RE (2013) Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 16(5):580–589CrossRefPubMedGoogle Scholar
  12. Doncaster CP, Jackson A, Watson RA (2013) Manipulated into giving: when parasitism drives apparent or incidental altruism. Proc R Soc Lond B Biol 280(1758):20130108CrossRefGoogle Scholar
  13. Feng XC, Guo WQ, Yang SS, Zheng HS, Du JS, Wu QL, Ren NQ (2014) Possible causes of excess sludge reduction adding metabolic uncoupler, 3,3′,4′,5-tetrachlorosalicylanilide (TCS), in sequence batch reactors. Bioresour Technol 173:96–103CrossRefPubMedGoogle Scholar
  14. Geng H, Belas R (2010a) Expression of tropodithietic acid biosynthesis is controlled by a novel autoinducer. J Bacteriol 192(17):4377–4387CrossRefPubMedPubMedCentralGoogle Scholar
  15. Geng H, Belas R (2010b) Molecular mechanisms underlying roseobacter-phytoplankton symbioses. Curr Opin Biotechnol 21(3):332–338CrossRefPubMedGoogle Scholar
  16. Gerardi MH (2006) Wastewater Bacteria. John Wiley & Sons, Inc., New YorkCrossRefGoogle Scholar
  17. Guo F, Zhang SH, Yu X, Wei B (2011) Variations of both bacterial community and extracellular polymers: the inducements of increase of cell hydrophobicity from biofloc to aerobic granule sludge. Bioresour Technol 102(11):6421–6428CrossRefPubMedGoogle Scholar
  18. Hancock L, Goff L, Lane C (2010) Red algae lose key mitochondrial genes in response to becoming parasitic. Genome Biol Evol 2(1):897–910CrossRefPubMedPubMedCentralGoogle Scholar
  19. Helliwell KE, Wheeler GL, Leptos KC, Goldstein RE, Smith AG (2011) Insights into the evolution of vitamin B12 auxotrophy from sequenced algal genomes. Mol Biol Evol 28(10):2921–2933CrossRefPubMedGoogle Scholar
  20. Herbert D, Phipps PJ, Strange RE (1971) Chapter III chemical analysis of microbial cells. Elsevier LtdGoogle Scholar
  21. Huang W, Li B, Zhang C, Zhang Z, Lei Z, Lu B, Zhou B (2015) Effect of algae growth on aerobic granulation and nutrients removal from synthetic wastewater by using sequencing batch reactors. Bioresour Technol 179:187–192CrossRefPubMedGoogle Scholar
  22. Kanagasabhapathy M, Yamazaki G, Ishida A, Sasaki H, Nagata S (2009) Presence of quorum-sensing inhibitor-like compounds from bacteria isolated from the brown alga Colpomenia sinuosa. Lett Appl Microbiol 49(5):573–579CrossRefPubMedGoogle Scholar
  23. Kazamia E, Czesnick H, Nguyen TTV, Croft MT, Sherwood E, Sasso S, Hodson SJ, Warren MJ, Smith AG (2012) Mutualistic interactions between vitamin B12-dependent algae and heterotrophic bacteria exhibit regulation. Environ Microbiol 14(6):1466–1476CrossRefPubMedGoogle Scholar
  24. Kim BH, Kang Z, Ramanan R, Choi JE, Cho DH, Oh HM, Kim HS (2014) Nutrient removal and biofuel production in high rate algal pond using real municipal wastewater. J Microbiol Biotechnol 24(8):1123–1132CrossRefPubMedGoogle Scholar
  25. Küpper FC, Müller DG, Peters AF, Kloareg B, Potin P (2002) Oligoalginate recognition and oxidative burst play a key role in natural and induced resistance of sporophytes of Laminariales. J Chem Ecol 28(10):2057–2081CrossRefPubMedGoogle Scholar
  26. Li YC, Zhu JR (2014) Role of N-acyl homoserine lactone (AHL)-based quorum sensing (QS) in aerobic sludge granulation. Appl Microbiol Biotechnol 98(17):7623–7632CrossRefPubMedGoogle Scholar
  27. Li Y, Lv J, Chen Z, Hao W, Wang Y, Zhu J (2014) Performance and role of N-acyl-homoserine lactone (AHL)-based quorum sensing (QS) in aerobic granules. J Environ Sci 26(8):1615–1621CrossRefGoogle Scholar
  28. Lin L, Fan H, Liu Y, Liu C, Xu H (2017) Development of algae-bacteria granular consortia in photo-sequencing batch reactor. Bioresour Technol 232:64–71CrossRefGoogle Scholar
  29. Lv J, Wang Y, Chen Z, Li Y, Wen H, Zhu J (2014) The effect of quorum sensing and extracellular proteins on the microbial attachment of aerobic granular activated sludge. Bioresour Technol 152(1):53–58CrossRefPubMedGoogle Scholar
  30. Manefield M, Turner SL (2002) Quorum sensing in context: out of molecular biology and into microbial ecology. Microbiology 148:3762–3764CrossRefPubMedGoogle Scholar
  31. Mangwani N, Kumari S, Das S (2016) Effect of synthetic N-acylhomoserine lactones on cell-cell interactions in marine Pseudomonas and biofilm mediated degradation of polycyclic aromatic hydrocarbons. Chem Eng J 302(11):172–186CrossRefGoogle Scholar
  32. Mitsutani A, Yamasaki I, Kitaguchi H, Kato J, Ueno S, Ishida Y (2001) Analysis of algicidal proteins of a diatom-lytic marine bacterium Pseudoalteromonas sp. strain A25 by two-dimensional electrophoresis. Phycologia 40(3):286–291CrossRefGoogle Scholar
  33. Natrah FM, Defoirdt T, Sorgeloos P, Bossier P (2011) Disruption of bacterial cell-to-cell communication by marine organisms and its relevance to aquaculture. Mar Biotechnol 13(2):109–126CrossRefPubMedGoogle Scholar
  34. Niu C, Clemmer KM, Bonomo RA, Rather PN (2008) Isolation and characterization of an autoinducer synthase from Acinetobacter baumannii. J Bacteriol 190(9):3386–3392CrossRefPubMedPubMedCentralGoogle Scholar
  35. Ponnusamy K, Paul D, Kweon J (2009) Inhibition of quorum sensing mechanism and Aeromonas hydrophila biofilm formation by vanillin. Environ Eng Sci 26(8):1359–1363CrossRefGoogle Scholar
  36. Ramanan R, Kang Z, Kim BH, Cho DH, Jin L, Oh HM, Kim HS (2015) Phycosphere bacterial diversity in green algae reveals an apparent similarity across habitats. Algal Res 8:140–144CrossRefGoogle Scholar
  37. Ramanan R, Kim BH, Cho DH, Oh HM, Kim HS (2016) Algae-bacteria interactions: evolution, ecology and emerging applications. Biotechnol Adv 34(1):14–29CrossRefPubMedGoogle Scholar
  38. Ren TT, Yu HQ, Li XY (2010) The quorum-sensing effect of aerobic granules on bacterial adhesion, biofilm formation, and sludge granulation. Appl Microbiol Biotechnol 88(3):789–797CrossRefPubMedGoogle Scholar
  39. Shrout JD, Nerenberg R (2012) Monitoring bacterial twitter: does quorum sensing determine the behavior of water and wastewater treatment biofilms? Environ Sci Technol 46(4):1995–2005CrossRefPubMedGoogle Scholar
  40. Su Y, Mennerich A, Urban B (2012) Synergistic cooperation between wastewater-born algae and activated sludge for wastewater treatment: influence of algae and sludge inoculation ratios. Bioresour Technol 105:67–73CrossRefPubMedGoogle Scholar
  41. Tan CH, Kai SK, Chao X, Tay M, Yan Z, Williams R, Ng WJ, Rice SA, Kjelleberg S (2014) The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules. ISME J 8(6):1186–1197CrossRefPubMedPubMedCentralGoogle Scholar
  42. Teplitski M, Rajamani S (2011) Signal and nutrient exchange in the interactions between soil algae and Bacteria. Springer, BerlinCrossRefGoogle Scholar
  43. Tiron O, Bumbac C, Patroescu IV, Badescu VR, Postolache C (2015) Granular activated algae for wastewater treatment. Water Sci Technol 71(6):832–839CrossRefPubMedGoogle Scholar
  44. Unnithan VV, Unc A, Smith GB (2014) Mini-review: a priori considerations for bacteria–algae interactions in algal biofuel systems receiving municipal wastewaters. Algal Res 4(2):35–40CrossRefGoogle Scholar
  45. Walter WG (1971) Standard methods for the examination of water and wastewater. APHAGoogle Scholar
  46. Wan C, Lee DJ, Yang X, Wang Y, Wang X, Liu X (2015) Calcium precipitate induced aerobic granulation. Bioresour Technol 176:32–37CrossRefPubMedGoogle Scholar
  47. Wilén BM, Jin B, Lant P (2003) The influence of key chemical constituents in activated sludge on surface and flocculating properties. Water Res 37(9):2127–2139CrossRefPubMedGoogle Scholar
  48. Williams P, Winzer K, Chan WC, Cámara M (2007) Look who’s talking: communication and quorum sensing in the bacterial world. Philos Trans R Soc B 362(1483):1119–1134CrossRefGoogle Scholar
  49. Wu LJ, Li AJ, Hou BL, Li MX (2017) Exogenous addition of cellular extract N -acyl-homoserine-lactones accelerated the granulation of autotrophic nitrifying sludge. Int Biodeterior Biodegrad 118:119–125CrossRefGoogle Scholar
  50. Zhang B, Lens PNL, Shi WX, Zhang RJ, Zhang ZQ, Guo Y, Bao X, Cui FY (2018) Enhancement of aerobic granulation and nutrient removal by an algal-bacterial consortium in a lab-scale photobioreactor. Chem Eng J 334:2373–2382CrossRefGoogle Scholar
  51. Zhou D, Zhang C, Fu L, Xu L, Cui X, Li Q, Crittenden JC (2017) Responses of the microalga Chlorophyta sp. to bacterial quorum sensing molecules (N-acylhomoserine lactones): aromatic protein-induced self-aggregation. Environ Sci Technol 51(6):3490–3498CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Bing Zhang
    • 1
  • Piet N. L. Lens
    • 2
  • Wenxin Shi
    • 1
  • Ruijun Zhang
    • 1
  • Zhiqiang Zhang
    • 1
  • Yuan Guo
    • 1
  • Xian Bao
    • 1
  • Fuyi Cui
    • 1
    • 3
  1. 1.State Key Laboratory of Urban Water Resource and EnvironmentHarbin Institute of TechnologyHarbinChina
  2. 2.UNESCO-IHEDelftThe Netherlands
  3. 3.College of Urban Construction and Environmental EngineeringChongqing UniversityChongqingChina

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