Whole genome sequencing enables the characterization of BurI, a LuxI homologue of Burkholderia cepacia strain GG4.
Quorum sensing is a mechanism for regulating proteobacterial gene expression in response to changes in cell population. In proteobacteria, N-acyl homoserine lactone (AHL) appears to be the most widely used signalling molecules in mediating, among others, the production of extracellular virulence factors for survival. In this work, the genome of B. cepacia strain GG4, a plasmid-free strain capable of AHL synthesis was explored. In silico analysis of the 6.6 Mb complete genome revealed the presence of a LuxI homologue which correspond to Type I quorum sensing. Here, we report the molecular cloning and characterization of this LuxI homologue, designated as BurI. This 609 bp gene was cloned and overexpressed in Escherichia coli BL21(DE3). The purified protein was approximately 25 kDa and is highly similar to several autoinducer proteins of the LuxI family among Burkholderia species. To verify the AHL synthesis activity of this protein, high resolution liquid chromatography-mass spectrometry analysis revealed the production of 3-oxo-hexanoylhomoserine lactone, N-octanoylhomoserine lactone and 3-hydroxy-octanoylhomoserine lactone from induced E. coli BL21 harboring the recombinant BurI. Our data show, for the first time, the cloning and characterization of the LuxI homologue from B. cepacia strain GG4 and confirmation of its AHL synthesis activity.
Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya , Kuala Lumpur , Malaysia.
Kah Yan
KY
How
Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya , Kuala Lumpur , Malaysia.
Kar Wai
KW
Hong
Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya , Kuala Lumpur , Malaysia.
Kok-Gan
KG
Chan
AHL synthase; Burkholderia cepacia; Liquid chromatograhy mass spectrometry (LC-MS); N-acylhomoserine lactone (AHL); Protein expression; Quorum sensing