RESUMEN
We report the complete genome sequence of Pseudomonas putida strain TS312, in the class of Gammaproteobacteria The strain, isolated from a paper mill, harbors the hdtS gene, encoding N-acyl-homoserine lactone synthase. Deciphering the genome contributes to revealing the mechanisms of quorum sensing and associated biofilm formation in engineered systems.
RESUMEN
N-Acylhomoserine lactone (AHL)-degrading enzyme is identified from the various environments and applied for quorum-sensing inhibition. In this study, we isolated two AHL-degrading strains, Sphingopyxis sp. EG6 and FD7, from the industrial cooling water samples. When the eight Sphingopyxis type strains were checked for the AHL-degrading activity, two strains, Sphingopyxis alaskensis DSM 13593 and Sphingopyxis bauzanensis DSM 22271, showed high AHL-degrading activity. The complete genome sequences of EG6 and FD7 revealed the presence of gene homolog of qsdS, which encodes AHL-lactonase in Sphingomonas ursincola. The qsdS gene is seated between putative gene homologs involved in 3-isopropylmalate dehydratase large (leuC2) and small (leuD) subunits in the genome of EG6, FD7, DSM 13593, and DSM 22271, but completely disappeared between leuC2 and leuD in the genome sequences of Sphingopyxis type strains without AHL-degrading activity. Purified His-tagged QsdS showed high AHL-degrading activity and catalyzed AHL ring opening by hydrolyzing lactones. In addition, heterologous expression of qsdS in Pseudomonas aeruginosa resulted in reduction of biofilm formation. These results suggested that the AHL-degrading activity in Sphingopyxis is useful as an effective agent for biofilm inhibition.
Asunto(s)
Hidrolasas de Éster Carboxílico , Lactonas/metabolismo , Sphingomonadaceae/enzimología , Sphingomonadaceae/genética , Secuencia de Aminoácidos , Secuencia de Bases , Biopelículas/crecimiento & desarrollo , Hidrolasas de Éster Carboxílico/análisis , Hidrolasas de Éster Carboxílico/genética , Hidrolasas de Éster Carboxílico/metabolismo , Mapeo Cromosómico , Clonación Molecular , Genoma Bacteriano , Filogenia , Pseudomonas aeruginosa/enzimología , Pseudomonas aeruginosa/genética , Percepción de Quorum/genética , Análisis de Secuencia de ADN , Sphingomonadaceae/fisiología , Sphingomonas/enzimología , Sphingomonas/genéticaRESUMEN
Biofilm formation by bacteria is one of the main causes of fouling in industrial cooling water systems. In many gram-negative bacteria, biofilm formation is regulated by N-acyl-homoserine lactone (AHL)-mediated quorum sensing. In this study, we isolated three AHL-degrading bacteria from cooling water systems and identified them as Sphingomonas ursincola. The draft genome sequence of S. ursincola A1 revealed the presence of an AHL-degrading gene homolog, designated qsdS. The qsdS region was also amplified by PCR from the genomes of the other two S. ursincola strains, SF1 and SF8. Escherichia coli DH5α harboring a QsdS-expressing plasmid showed high degradative activity against AHLs with short and 3-oxo-substituted acyl chains. High-performance liquid chromatography analysis revealed that QsdS is an AHL lactonase, an enzyme that catalyzes AHL ring opening. Furthermore, heterologous expression of QsdS in Pseudomonas aeruginosa PAO1 resulted in degradation of endogenous AHLs and interfered with the quorum-sensing-regulated phenotype.