Metagenome-Derived Clones Encoding Two Novel Lactonase Family Proteins Involved in Biofilm Inhibition in Pseudomonas aeruginosa

Here we report the isolation and characterization of three metagenome-derived clones that interfere with bacterial quorum sensing and degrade N-(3-oxooctanoyl)-L-homoserine lactone (3-oxo-C₈-HSL). By using a traI-lacZ gene fusion, the metagenome-derived clones were identified from a soil DNA library...

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Bibliographic Details
Published in:Applied and Environmental Microbiology 2009-01, Vol.75 (1), p.224-233
Main Authors: Schipper, C, Hornung, C, Bijtenhoorn, P, Quitschau, M, Grond, S, Streit, W.R
Format: Article
Language:English
Subjects:
4-Butyrolactone - analogs & derivatives
4-Butyrolactone - metabolism
Amino acids
Bacterial proteins
Biofilms
Biofilms - growth & development
Carboxylic Ester Hydrolases - genetics
Carboxylic Ester Hydrolases - metabolism
Cloning
Cloning, Molecular
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Enzymology and Protein Engineering
Gene Library
Mass spectrometry
Microbiology
Molecular Sequence Data
Open Reading Frames
Pseudomonas aeruginosa
Pseudomonas aeruginosa - enzymology
Pseudomonas aeruginosa - genetics
Sequence Analysis, DNA
Soil Microbiology
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Summary:Here we report the isolation and characterization of three metagenome-derived clones that interfere with bacterial quorum sensing and degrade N-(3-oxooctanoyl)-L-homoserine lactone (3-oxo-C₈-HSL). By using a traI-lacZ gene fusion, the metagenome-derived clones were identified from a soil DNA library and analyzed. The open reading frames linked to the 3-oxo-C₈-HSL-degrading activities were designated bpiB01, bpiB04, and bpiB07. While the BpiB07 protein was similar to a known lactonase, no significant similarities were observed for the BpiB01 and BpiB04 proteins or the deduced amino acid sequences. High-performance liquid chromatography-mass spectrometry analyses confirmed that the identified genes encode novel lactone-hydrolyzing enzymes. The original metagenome-derived clones were expressed in Pseudomonas aeruginosa and employed in motility and biofilm assays. All clones were able to reproducibly inhibit motility in P. aeruginosa. Furthermore, these genes clearly inhibited biofilm formation in P. aeruginosa when expressed in P. aeruginosa PAO1. Thus, this is the first study in which metagenome-derived proteins have been expressed in P. aeruginosa to successfully inhibit biofilm formation.
ISSN:0099-2240
1098-5336
1098-6596
DOI:10.1128/AEM.01389-08