Applied Microbiology and Biotechnology

, Volume 103, Issue 21–22, pp 9169–9180 | Cite as

Role of extracellular polymeric substances in biofilm formation by Pseudomonas stutzeri strain XL-2

  • Xue Song Ding
  • Bin ZhaoEmail author
  • Qiang An
  • Meng Tian
  • Jin Song Guo
Environmental biotechnology


Pseudomonas stutzeri strain XL-2 exhibited significant performance on biofilm formation. Extracellular polymeric substances (EPS) secreted by strain XL-2 were characterized by colorimetry and Fourier transform infrared (FT-IR) spectroscopy. The biofilm growth showed a strong positive correlation (rP=0.96, P<0.01) to extracellular protein content, but no correlation to exopolysaccharide content. Hydrolyzing the biofilm with proteinase K caused a significant decrease in biofilm growth (t=3.7, P<0.05), whereas the changes in biofilm growth were not significant when the biofilm was hydrolyzed by α-amylase and β-amylase, implying that proteins rather than polysaccharides played the dominant role in biofilm formation. More specifically, confocal laser scanning microscopy (CLSM) revealed that the extracellular proteins were tightly bound to the cells, resulting in the cells with EPS presenting more biofilm promotion protein secondary structures, such as three-turn helices, β-sheet, and α-helices, than cells without EPS. Both bio-assays and quantitative analysis demonstrated that strain XL-2 produced signal molecules of N-acylhomoserine lactones (AHLs) during biofilm formation process. The concentrations of C6-HLS and C6-oxo-HLS were both significantly positively correlated with protein contents (P<0.05). Dosing exogenous C6-HLS and C6-oxo-HLS also resulted in the increase in protein content. Therefore, it was speculated that C6-HLS and C6-oxo-HLS released by strain XL-2 could up-regulate the secretion of proteins in EPS, and thus promote the formation of biofilm.


Biofilm formation Extracellular polymeric substances (EPS) Extracellular proteins N-Acylhomoserine lactones (AHLs) Pseudomonas stutzeri 



This work was supported by the National Natural Science Foundation of China (grant no. 51208534) and Technical Innovation and Application Demonstration Project of CQ CSTC (grant no. cstc2018jscx-msybX0308).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of the Three Gorges Reservoir Region’s Eco-Environment, Ministry of EducationChongqing UniversityChongqingPeople’s Republic of China

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