Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family
Background Members of the genus Novosphingobium have been isolated from a variety of environmental niches. Although genomics analyses have suggested the presence of genes associated with quorum sensing signal production e.g., the N-acyl-homoserine lactone (AHL) synthase (luxI) homologs in various No...
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PeerJ Inc.
2016-08-01
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| author | Han Ming Gan Lucas K. Dailey Nigel Halliday Paul Williams André O. Hudson Michael A. Savka |
| author_facet | Han Ming Gan Lucas K. Dailey Nigel Halliday Paul Williams André O. Hudson Michael A. Savka |
| author_sort | Han Ming Gan |
| collection | DOAJ |
| description | Background Members of the genus Novosphingobium have been isolated from a variety of environmental niches. Although genomics analyses have suggested the presence of genes associated with quorum sensing signal production e.g., the N-acyl-homoserine lactone (AHL) synthase (luxI) homologs in various Novosphingobium species, to date, no luxI homologs have been experimentally validated. Methods In this study, we report the draft genome of the N-(AHL)-producing bacterium Novosphingobium subterraneum DSM 12447 and validate the functions of predicted luxI homologs from the bacterium through inducible heterologous expression in Agrobacterium tumefaciens strain NTL4. We developed a two-dimensional thin layer chromatography bioassay and used LC-ESI MS/MS analyses to separate, detect and identify the AHL signals produced by the N. subterraneum DSM 12447 strain. Results Three predicted luxI homologs were annotated to the locus tags NJ75_2841 (NovINsub1), NJ75_2498 (NovINsub2), and NJ75_4146 (NovINsub3). Inducible heterologous expression of each luxI homologs followed by LC-ESI MS/MS and two-dimensional reverse phase thin layer chromatography bioassays followed by bioluminescent ccd camera imaging indicate that the three LuxI homologs are able to produce a variety of medium-length AHL compounds. New insights into the LuxI phylogeny was also gleemed as inferred by Bayesian inference. Discussion This study significantly adds to our current understanding of quorum sensing in the genus Novosphingobium and provide the framework for future characterization of the phylogenetically interesting LuxI homologs from members of the genus Novosphingobium and more generally the family Sphingomonadaceae. |
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| last_indexed | 2024-03-09T08:00:30Z |
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| spelling | doaj.art-5ce79b50424e4f4d8922dbf5566d889d2023-12-03T00:46:41ZengPeerJ Inc.PeerJ2167-83592016-08-014e233210.7717/peerj.2332Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae familyHan Ming Gan0Lucas K. Dailey1Nigel Halliday2Paul Williams3André O. Hudson4Michael A. Savka5School of Science, Monash University Malaysia, Bandar Sunway, Selangor, MalaysiaThomas H. Gosnell School of School of Life Sciences, Rochester Institute of Technology, Rochester, NY, USASchool of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UKSchool of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UKThomas H. Gosnell School of School of Life Sciences, Rochester Institute of Technology, Rochester, NY, USAThomas H. Gosnell School of School of Life Sciences, Rochester Institute of Technology, Rochester, NY, USABackground Members of the genus Novosphingobium have been isolated from a variety of environmental niches. Although genomics analyses have suggested the presence of genes associated with quorum sensing signal production e.g., the N-acyl-homoserine lactone (AHL) synthase (luxI) homologs in various Novosphingobium species, to date, no luxI homologs have been experimentally validated. Methods In this study, we report the draft genome of the N-(AHL)-producing bacterium Novosphingobium subterraneum DSM 12447 and validate the functions of predicted luxI homologs from the bacterium through inducible heterologous expression in Agrobacterium tumefaciens strain NTL4. We developed a two-dimensional thin layer chromatography bioassay and used LC-ESI MS/MS analyses to separate, detect and identify the AHL signals produced by the N. subterraneum DSM 12447 strain. Results Three predicted luxI homologs were annotated to the locus tags NJ75_2841 (NovINsub1), NJ75_2498 (NovINsub2), and NJ75_4146 (NovINsub3). Inducible heterologous expression of each luxI homologs followed by LC-ESI MS/MS and two-dimensional reverse phase thin layer chromatography bioassays followed by bioluminescent ccd camera imaging indicate that the three LuxI homologs are able to produce a variety of medium-length AHL compounds. New insights into the LuxI phylogeny was also gleemed as inferred by Bayesian inference. Discussion This study significantly adds to our current understanding of quorum sensing in the genus Novosphingobium and provide the framework for future characterization of the phylogenetically interesting LuxI homologs from members of the genus Novosphingobium and more generally the family Sphingomonadaceae.https://peerj.com/articles/2332.pdfAcyl-homoserine lactonesLuxIRQuorum-sensingTwo-dimensional thin-layer chromatographyNovosphingobiumPhylogenetic |
| spellingShingle | Han Ming Gan Lucas K. Dailey Nigel Halliday Paul Williams André O. Hudson Michael A. Savka Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family PeerJ Acyl-homoserine lactones LuxIR Quorum-sensing Two-dimensional thin-layer chromatography Novosphingobium Phylogenetic |
| title | Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family |
| title_full | Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family |
| title_fullStr | Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family |
| title_full_unstemmed | Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family |
| title_short | Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family |
| title_sort | genome sequencing assisted identification and the first functional validation of n acyl homoserine lactone synthases from the sphingomonadaceae family |
| topic | Acyl-homoserine lactones LuxIR Quorum-sensing Two-dimensional thin-layer chromatography Novosphingobium Phylogenetic |
| url | https://peerj.com/articles/2332.pdf |
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