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Microbiology 146 (2000), 2469-2480
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Microbiology (2000), 146, 2469-2480.
© 2000 Society for General Microbiology

Pseudomonas: Biology and Diversity

The biocontrol strain Pseudomonas fluorescens F113 produces the Rhizobium small bacteriocin, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone, via HdtS, a putative novel N-acylhomoserine lactone synthase

Bridget E. Laue1, Yan Jiang1, Siri Ram Chhabra1, Sinead Jacob2, Gordon S. A. B. Stewartb,1, Andrea Hardman1, J. Allan Downie3, Fergal O’Gara2 and Paul Williams1

School of Pharmaceutical Sciences, University of Nottingham, Nottingham NG7 2RD, UK1
Biomerit Research Centre, Department of Microbiology, National University of Ireland, Cork, Ireland2
John Innes Centre, Colney, Norwich NR4 7UH, UK3

Author for correspondence: Paul Williams. Tel: +44 115 9515047. Fax: +44 115 8466296. e-mail: paul.williams{at}nottingham.ac.uk

Several different species of Pseudomonas produce N-acylhomoserine lactones (AHLs), quorum-sensing signal molecules which are involved in the cell-density-dependent control of secondary metabolite and virulence gene expression. When Pseudomonas fluorescens F113 was cross-streaked against AHL biosensors capable of sensitively detecting either short (C4–C8) or long (C10–C14) acyl chain AHLs, no activity was detectable. However, by extracting cell-free stationary-phase culture supernatants with dichloromethane followed by reverse-phase HPLC, three distinct fractions were obtained capable of activating the AHL biosensors. Three AHLs were subsequently characterized using high-resolution MS and chemical synthesis. These were (i) N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone (3OH,C14:1-HSL), a molecule previously known as the Rhizobium leguminosarum small bacteriocin as a consequence of its growth inhibitory properties, (ii) N-decanoylhomoserine lactone (C10-HSL) and (iii) N-hexanoylhomoserine lactone (C6-HSL). A gene (hdtS) capable of directing synthesis of all three P. fluorescens AHLs in Escherichia coli was cloned and sequenced. In vitro transcription/translation of hdtS yielded a protein of approximately 33 kDa capable of directing the synthesis of 3OH,C14:1-HSL, C10-HSL and C6-HSL in E. coli. HdtS does not belong to either of the known AHL synthase families (LuxI or LuxM) and is related to the lysophosphatidic acid acyltransferase family. HdtS may therefore constitute a member of a third protein family capable of AHL biosynthesis.

Keywords: Pseudomonas fluorescens, N-acylhomoserine lactone synthase, Rhizobium, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone, acyltransferase

Abbreviations: AHL, N-acylhomoserine lactone; DCM, dichloromethane; C6-HSL, N-hexanoylhomoserine lactone; C10-HSL, N-decanoylhomoserine lactone; 3OH,C14:1-HSL, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone; PDA, photodiode array

The GenBank accession number for the sequence determined in this work is AF286536.

b Deceased.




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