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. 2011 Dec;1(4):239-245.
doi: 10.1007/s13205-011-0029-1. Epub 2011 Oct 14.

Identification of N-acyl-l-homoserine lactones produced by non-pigmented Chromobacterium aquaticum CC-SEYA-1(T) and pigmented Chromobacterium subtsugae PRAA4-1(T)

P D Rekha, Chiu-Chung Young, A B Arun

Identification of N-acyl-l-homoserine lactones produced by non-pigmented Chromobacterium aquaticum CC-SEYA-1(T) and pigmented Chromobacterium subtsugae PRAA4-1(T)

P D Rekha et al. 3 Biotech. 2011 Dec.

Abstract

Many members of the genus Chromobacterium produce violacein, a characteristic purple pigment which is induced by small diffusible N-acyl homoserine lactones (AHL) quorum-sensing molecules. In this study, the production of AHL of the non-pigmented C. aquaticum CC-SEYA-1(T) and the pigmented C. subtsugae PRAA4-1(T) were determined by using a CV026 biosensor assay. The profile of AHL was identified from the extracts of stationary phase cultures using gas chromatography-mass spectroscopy (GC-MS) and thin layer chromatography (TLC). CV026 biosensor assay revealed that both the non-pigmented C. aquaticum CC-SEYA-1(T) and the pigmented C. subtsugae PRAA4-1(T) produced AHL molecules, which were identified, respectively, as N-octanoyl homoserine lactone (OHL) [also known as C-8 homoserine lactone (C8-HSL)] and N-hexanoyl homoserine lactone (HHL) [also known as C-6 homoserine lactone (C6-HSL)]. The pigment produced by C. subtsugae PRAA4-1(T) was similar to that of Chromobacterium violaceum ATCC12472(T) but no characteristic visible spectral peaks of the pigment were observed in the extracts of C. aquaticum CC-SEYA-1(T). In addition, C. aquaticum CC-SEYA-1(T) and C. subtsugae PRAA4-1(T) showed hemolytic activities.

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Figures

Fig. 1
Fig. 1
Biosensor plate bioassay showing the induction of violacein in C. violaceum CV026 biosensor strain by stationary phase cell-free supernatants of aC. aquaticum CC-SEYA-1T and bC. subtsugae PRAA4-1T
Fig. 2
Fig. 2
GC–MS chromatogram in SIM mode at m/z 143 of standard AHL compounds with their retention times. Synthetic AHLs used are, N-butanoyl (BHL), N-hexanoyl (HHL), N-octanoyl (OHL), N-decanoyl (DHL), N-dodecanoyl (dDHL), and N-tetradecanoyl (tDHL) homoserine lactones
Fig. 3
Fig. 3
GC–MS chromatogram in SIM mode at m/z 143 of AHL compounds from cell-free supernatants extracted in organic solvents of aC. aquaticum CC-SEYA-1T and bC. subtsugae PRAA4-1T with their retention times
Fig. 4
Fig. 4
Zones of clearance around colonies of aC. violaceum ATCC 12472, bC. aquaticum CC-SEYA-1T, and cC. subtsugae PRAA4-1T on sheep blood agar indicating hemolysis
See this image and copyright information in PMC

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