title: A novel metagenomic short-chain dehydrogenase/reductase attenuates Pseudomonas aeruginosa biofilm formation and virulence on Caenorhabditis elegans.
abstract: In Pseudomonas aeruginosa, the expression of a number of virulence factors, as well as biofilm formation, are controlled by quorum sensing (QS). N-Acylhomoserine lactones (AHLs) are an important class of signaling molecules involved in bacterial QS and in many pathogenic bacteria infection and host colonization are AHL-dependent. The AHL signaling molecules are subject to inactivation mainly by hydrolases (Enzyme Commission class number EC 3) (i.e. N-acyl-homoserine lactonases and N-acyl-homoserine-lactone acylases). Only little is known on quorum quenching mechanisms of oxidoreductases (EC 1). Here we report on the identification and structural characterization of the first NADP-dependent short-chain dehydrogenase/reductase (SDR) involved in inactivation of N-(3-oxo-dodecanoyl)-L-homoserine lactone (3-oxo-C(12)-HSL) and derived from a metagenome library. The corresponding gene was isolated from a soil metagenome and designated bpiB09. Heterologous expression and crystallographic studies established BpiB09 as an NADP-dependent reductase. Although AHLs are probably not the native substrate of this metagenome-derived enzyme, its expression in P. aeruginosa PAO1 resulted in significantly reduced pyocyanin production, decreased motility, poor biofilm formation and absent paralysis of Caenorhabditis elegans. Furthermore, a genome-wide transcriptome study suggested that the level of lasI and rhlI transcription together with 36 well known QS regulated genes was significantly (≥10-fold) affected in P. aeruginosa strains expressing the bpiB09 gene in pBBR1MCS-5. Thus AHL oxidoreductases could be considered as potent tools for the development of quorum quenching strategies.
authors: Abteilung für Mikrobiologie und Biotechnologie, Biozentrum Klein Flottbek, Universität Hamburg, Hamburg, Germany.
authors: Patrick
authors: P
authors: Bijtenhoorn
authors: Hubert
authors: H
authors: Mayerhofer
authors: Jochen
authors: J
authors: Müller-Dieckmann
authors: Christian
authors: C
authors: Utpatel
authors: Christina
authors: C
authors: Schipper
authors: Claudia
authors: C
authors: Hornung
authors: Matthias
authors: M
authors: Szesny
authors: Stephanie
authors: S
authors: Grond
authors: Andrea
authors: A
authors: Thürmer
authors: Elzbieta
authors: E
authors: Brzuszkiewicz
authors: Rolf
authors: R
authors: Daniel
authors: Katja
authors: K
authors: Dierking
authors: Hinrich
authors: H
authors: Schulenburg
authors: Wolfgang R
authors: WR
authors: Streit
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000502:physiology*; D018441:Biofilms / Q000187:drug effects*; D017173:Caenorhabditis elegans / Q000382:microbiology*; D020869:Gene Expression Profiling; D056186:Metagenomics; D009249:NADP; D010088:Oxidoreductases / Q000302:isolation & purification* / Q000494:pharmacology* / Q000502:physiology; D011552:Pseudomonas Infections / Q000276:immunology; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity; D011710:Pyocyanine / Q000378:metabolism; D053038:Quorum Sensing / Q000187:drug effects; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D009249:NADP; D011710:Pyocyanine; D010088:Oxidoreductases