title: Development of bioreporter assays for the detection of bioavailability of long-chain alkanes based on the marine bacterium Alcanivorax borkumensis strain SK2.
abstract: Long-chain alkanes are a major component of crude oil and therefore potentially good indicators of hydrocarbon spills. Here we present a set of new bacterial bioreporters and assays that allow to detect long-chain alkanes. These reporters are based on the regulatory protein AlkS and the alkB1 promoter from Alcanivorax borkumensis SK2, a widespread alkane degrader in marine habitats. Escherichia coli cells with the reporter construct reacted strongly to octane in short-term (6 h) aqueous suspension assays but very slightly only to tetradecane, in line with what is expected from its low water solubility. In contrast, long-term assays (up to 5 days) with A. borkumensis bioreporters showed strong induction with tetradecane and crude oil. Gel-immobilized A. borkumensis reporter cells were used to demonstrate tetradecane and crude oil bioavailability at a distance from a source. Alcanivorax borkumensis bioreporters induced fivefold more rapid and more strongly when allowed physical contact with the oil phase in standing flask assays, suggesting a major contribution of adhered cells to the overall reporter signal. Using the flask assays we further demonstrated the effect of oleophilic nutrients and biosurfactants on oil availability and degradation by A. borkumensis. The fluorescence signal from flask assays could easily be captured with a normal digital camera, making such tests feasible to be carried out on, e.g. marine oil responder vessels in case of oil accidents.
authors: Department of Fundamental Microbiology, University of Lausanne, 1015 Lausanne, Switzerland.
authors: Rekha
authors: R
authors: Kumari
authors: Robin
authors: R
authors: Tecon
authors: Siham
authors: S
authors: Beggah
authors: Rebecca
authors: R
authors: Rutler
authors: J Samuel
authors: JS
authors: Arey
authors: Jan Roelof
authors: JR
authors: van der Meer
mesh_terms: D053521:Alcanivoraceae / Q000235:genetics / Q000378:metabolism*; D000473:Alkanes / Q000378:metabolism*; D001682:Biological Availability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D009795:Octanes / Q000378:metabolism*; D010578:Petroleum / Q000378:metabolism; D010957:Plasmids
chemical_list: D000473:Alkanes; D009795:Octanes; D010578:Petroleum; C024713:n-tetradecane; C026728:octane