A phospholipase B from Pseudomonas aeruginosa with activity towards endogenous phospholipids affects biofilm assembly

Biochim Biophys Acta Mol Cell Biol Lipids. 2022 Apr;1867(4):159101. doi: 10.1016/j.bbalip.2021.159101. Epub 2022 Jan 19.


Pseudomonas aeruginosa is a severe threat to immunocompromised patients due to its numerous virulence factors and biofilm-mediated multidrug resistance. It produces and secretes various toxins with hydrolytic activities including phospholipases. However, the function of intracellular phospholipases for bacterial virulence has still not been established. Here, we demonstrate that the hypothetical gene pa2927 of P. aeruginosa encodes a novel phospholipase B named PaPlaB. At reaction equilibrium, PaPlaB purified from detergent-solubilized membranes of E. coli released fatty acids (FAs) from sn-1 and sn-2 positions of phospholipids at the molar ratio of 51:49. PaPlaB in vitro hydrolyzed P. aeruginosa phospholipids reconstituted in detergent micelles and phospholipids reconstituted in vesicles. Cellular localization studies indicate that PaPlaB is a cell-bound PLA of P. aeruginosa and that it is peripherally bound to both membranes in E. coli, yet the active form was predominantly associated with the cytoplasmic membrane of E. coli. Decreasing the concentration of purified and detergent-stabilized PaPlaB leads to increased enzymatic activity, and at the same time triggers oligomer dissociation. We showed that the free FA profile, biofilm amount and architecture of the wild type and ΔplaB differ. However, it remains to be established how the PLB activity of PaPlaB is regulated by homooligomerisation and how it relates to the phenotype of the P. aeruginosa ΔplaB. This novel putative virulence factor contributes to our understanding of phospholipid degrading enzymes and might provide a target for new therapeutics against P. aeruginosa biofilms.

Keywords: Membrane protein; Oligomerization; Pathogen; Virulence factor; α/β-hydrolase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biofilms
  • Detergents / pharmacology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Lysophospholipase* / metabolism
  • Phospholipases / genetics
  • Phospholipases / metabolism
  • Phospholipids / metabolism
  • Pseudomonas aeruginosa*
  • Virulence Factors / genetics
  • Virulence Factors / metabolism


  • Detergents
  • Phospholipids
  • Virulence Factors
  • Phospholipases
  • Lysophospholipase