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Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family

Gan, Han Ming, Dailey, Lucas K., Halliday, Nigel, Williams, Paul, Hudson, Andre O. and Savka, Michael A. 2016, Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family, PeerJ, vol. 2016, no. 8, pp. 1-20, doi: 10.7717/peerj.2332.

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Title Genome sequencing-assisted identification and the first functional validation of N-acyl-homoserine-lactone synthases from the Sphingomonadaceae family
Author(s) Gan, Han MingORCID iD for Gan, Han Ming orcid.org/0000-0001-7987-738X
Dailey, Lucas K.
Halliday, Nigel
Williams, Paul
Hudson, Andre O.
Savka, Michael A.
Journal name PeerJ
Volume number 2016
Issue number 8
Article ID e2332
Start page 1
End page 20
Total pages 20
Publisher PeerJ
Place of publication London, Eng.
Publication date 2016
ISSN 2167-8359
Keyword(s) acyl-homoserine lactones
luxIR
quorum-sensing
two-dimensional thin-layer chromatography
Novosphingobium
phylogenetic
sphingomonadaceae
N-acyl-homoserine lactone synthases
whole genome sequencing
Summary 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 (NovI Nsub1 ), NJ75_2498 (NovI Nsub2 ), and NJ75_4146 (NovI Nsub3 ). Inducible heterologous expression of each luxI homologs followed by LC-ESI MS/MS and twodimensional 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.
Language eng
DOI 10.7717/peerj.2332
Field of Research 060408 Genomics
060503 Microbial Genetics
HERDC Research category C1.1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2016, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30101941

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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.