Heterologous expression of human paraoxonases in Pseudomonas aeruginosa inhibits biofilm formation and decreases antibiotic resistance
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Quorum sensing (QS) regulates virulence and biofilm formation in Pseudomonas aeruginosa and other medically relevant bacteria. Human paraoxonases (hPONs) are a family of closely related enzymes with multiple functions, including inactivation of the QS signal molecule in P. aeruginosa. However, there is no direct evidence to show the functions of hPONs on biofilm formation and antibiotic resistance in P. aeruginosa. In the present study, hPONs (hPON1, hPON2, and hPON3) genes were respectively cloned into the pMEKm12 shuttle vector and transformed into P. aeruginosa strain PAO1. Expression of the three recombinant proteins was confirmed by Western blotting, and growth of the recombinant strains was not affected by the hPONs gene expression. Biofilm formation and antibiotics resistance of the hPONs recombinant strains were analyzed. Our results showed that biofilm formation was significantly inhibited in all of the three hPONs recombinant strains. Interestingly, this inhibition can be reverted by addition of the corresponding hPONs polyclonal antibodies in the culture media, further indicating that the inhibition of biofilm formation was due to hPONs protein expression. In addition, we also demonstrated that hPONs expression decreased resistance of P. aeruginosa to gentamicin and ceftazidima, two antibiotics clinically used for the treatment of P. aeruginosa infection.
KeywordsPseudomonas aeruginosa Biofilm Antibiotic resistance Human paraoxonases Quorum sensing
This research was supported by a grant from the National Natural Science Foundation of China (#30470997). We thank Prof. Joseph Zabner, University of Iowa College of Medicine, for providing us with the adenovirus plasmids; Prof. Jun Zhu, Nanjing Agricultural University, for the Agrobacterium bioassay strain. We also thank the Animal Experimental Centre of Wuhan Institute of Virology for excellent technical assistance in the preparation of the hPONs polyclonal antibodies.
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