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Applied and Environmental Microbiology, January 2009, p. 224-233, Vol. 75, No. 1
0099-2240/09/$08.00+0     doi:10.1128/AEM.01389-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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

C. Schipper,¹ C. Hornung,¹ P. Bijtenhoorn,¹ M. Quitschau,² S. Grond,² and W. R. Streit^1*

Abteilung für Mikrobiologie und Biotechnologie, Biozentrum Klein Flottbek, University of Hamburg, Ohnhorststr. 18, D-22609 Hamburg, Germany,¹ Institut für Organische und Biomolekulare Chemie, University of Göttingen, Tammannstrasse 2, 37077 Göttingen, Germany²

Received 20 June 2008/ Accepted 30 October 2008

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.

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* Corresponding author. Mailing address: Biozentrum Klein Flottbek-Mikrobiologie, University of Hamburg, Ohnhorststr. 18, D-22609 Hamburg, Germany. Phone: 49-40-428-16-463. Fax: 49-40-428-16-459. E-mail: wolfgang.streit{at}uni-hamburg.de

Published ahead of print on 7 November 2008.

Supplemental material for this article may be found at http://aem.asm.org/.

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Applied and Environmental Microbiology, January 2009, p. 224-233, Vol. 75, No. 1
0099-2240/09/$08.00+0     doi:10.1128/AEM.01389-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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