Chromosomally located gene fusions constructed in Acinetobacter sp. ADP1 for the detection of salicylate.
Acinetobacter sp. ADP1 is a common soil-associated bacterium with high natural competency, allowing it to efficiently integrate foreign DNA fragments into its chromosome. This property was exploited to engineer salicylate-inducible luxCDABE and green fluorescent protein (GFP) variants of Acinetobacter sp. ADP1. Specifically, Acinetobacter sp. ADPWH_lux displayed the higher sensitivity when comparing the two variants (minimum detection c. 0.5-1 microM salicylate) and a faster turnover of the lux marker gene, making it suitable for whole-cell luminescence assays of salicylate concentration. In contrast, the longer maturation and turnover times of the GFP protein make the Acinetobacter sp. ADPWH_gfp variant more suited to applications involving whole-cell imaging of the presence of salicylate. The sensitivity of the luxCDABE variant was demonstrated by assaying salicylate production in naphthalene-degrading cultures. Assays using ADPWH_lux specifically mapped the kinetics of salicylate production from naphthalene and were similar to that observed by high-performance liquid chromatography (HPLC) data. However, ADPWH_lux exhibited the higher sensitivity, when compared with HPLC, for detecting salicylate production during the first 24 h of naphthalene metabolism. These data demonstrate that the engineered Acinetobacter variants have significant potential for salicylate detection strategies in laboratory and field studies, especially in scenarios where genetic stability of the construct is required for in situ monitoring.
Huang
Wei E
WE
Molecular Microbial Ecology, CEH-Oxford, Mansfield Road, Oxford OX1 3SR, UK.
Wang
Hui
H
Zheng
Hongjun
H
Huang
Linfeng
L
Singer
Andrew C
AC
Thompson
Ian
I
Whiteley
Andrew S
AS
D000150:Acinetobacter / Q000235:genetics* / Q000254:growth & development; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D001673:Biodegradation, Environmental; D015374:Biosensing Techniques; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004785:Environmental Pollutants / Q000032:analysis*; D005798:Genes, Bacterial / Q000235:genetics*; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D012459:Salicylates / Q000032:analysis*
D001426:Bacterial Proteins; D004785:Environmental Pollutants; D012459:Salicylates; D049452:Green Fluorescent Proteins