RT Journal Article SR Electronic(1) A1 Laue, Bridget E. A1 Jiang, Yan A1 Chhabra, Siri Ram A1 Jacob, Sinead A1 Stewart, Gordon S. A. B. A1 Hardman, Andrea A1 Downie, J. Allan A1 O’Gara, Fergal A1 Williams, Paul YR 2000 T1 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 synthaseThe GenBank accession number for the sequence determined in this work is AF286536. JF Microbiology, VO 146 IS 10 SP 2469 OP 2480 DO https://doi.org/10.1099/00221287-146-10-2469 PB Microbiology Society, SN 1465-2080, AB 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., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-146-10-2469