title: Chemical synthesis of (S)-4,5-dihydroxy-2,3-pentanedione, a bacterial signal molecule precursor, and validation of its activity in Salmonella typhimurium.
abstract: We describe an original, short, and convenient chemical synthesis of enantiopure (S)-4,5-dihydroxy-2,3-pentanedione (DPD), starting from commercial methyl (S)-(-)-2,2-dimethyl-1,3-dioxolane-4-carboxylate. DPD is the precursor of autoinducer (AI)-2, the proposed signal for bacterial interspecies communication. AI-2 is synthesized by many bacterial species in three enzymatic steps. The last step, a LuxS-catalyzed reaction, leads to the formation of DPD, which spontaneously cyclizes into AI-2. AI-2-like activity of the synthesized molecule was ascertained by the Vibrio harveyi bioassay. To further validate the biological activity of synthetic DPD and to explore its potential in studying DPD (AI-2)-mediated signaling, a Salmonella typhimurium luxS mutant was constructed. Expression of the AI-2 regulated lsr operon can be rescued in this luxS mutant by addition of synthetic DPD or genetic complementation. Biofilm formation by S. typhimurium has been reported to be defective in a luxS mutant, and this was confirmed in this study to test DPD for chemical complementation. However, biofilm formation of the luxS mutant cannot be restored by addition of DPD. In contrast, introduction of luxS under control of its own promoter complemented biofilm formation. Further results demonstrated that biofilm formation of the luxS mutant cannot be restored with luxS under control of the strong nptII promoter. This indicates that altering the intrinsic promoter activity of luxS affects Salmonella biofilm formation. Conclusively, we synthesized biologically active DPD. Using this chemical compound in combination with genetic approaches opens new avenues in studying AI-2-mediated signaling.
authors.affiliation: Centre of Microbial and Plant Genetics, Leuven, Belgium.
authors.forename: Sigrid C J
authors.initials: SC
authors.lastname: De Keersmaecker
authors.forename: Csaba
authors.initials: C
authors.lastname: Varszegi
authors.forename: Nadja
authors.initials: N
authors.lastname: van Boxel
authors.forename: Lothar W
authors.initials: LW
authors.lastname: Habel
authors.forename: Kristine
authors.initials: K
authors.lastname: Metzger
authors.forename: Ruth
authors.initials: R
authors.lastname: Daniels
authors.forename: Kathleen
authors.initials: K
authors.lastname: Marchal
authors.forename: Dirk
authors.initials: D
authors.lastname: De Vos
authors.forename: Jos
authors.initials: J
authors.lastname: Vanderleyden
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms / Q000187:drug effects; D013437:Carbon-Sulfur Lyases; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006714:Homoserine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D007783:Lactones; D009154:Mutation; D010420:Pentanes / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013237:Stereoisomerism
chemical_list: C493138:4,5-dihydroxy-2,3-pentanedione; D001426:Bacterial Proteins; D007783:Lactones; C091088:N-octanoylhomoserine lactone; D010420:Pentanes; D006714:Homoserine; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria