Physiological and toxicological characterization of an engineered whole-cell biosensor

Abstract

Bioluminescence-based bacterial biosensors are often reported as reliable and efficient tools for risk assessment and environmental monitoring. However, there are few data comparing the metabolism of genetically engineered strains to the corresponding wild type. A pollutant-degrading bacterium capable of mineralising 2,4-dichlorophenoxyacetic acid (2,4-D), Burkholderia sp. strain RASC c2, was genetically engineered to produce light constitutively and tested for assessing the main causes of biodegradation constraint affected by growth rates, toxicity, bioavailability and metal speciation in complex environments. This research focuses on such aspects by characterizing two pollutant-degrading isolates, the wild type and the genetically engineered biosensor ( lux-marked). Degradation and growth rates of both isolates were assessed with different concentrations of 2,4-D as the sole carbon source. Kinetic rates were affected by initial concentration of substrate and isolates showed distinct growth rates at different 2,4-D concentrations. Toxic effects of zinc and copper were also comparatively assessed using a dehydrogenase assay and light output. The isolates were sensitive to both metals and at similar EC 50 values. Therefore, bioluminescence response of the lux-marked isolate accurately reflected the toxic response of the parental organism towards zinc and copper, making it an ideal test-organism for assessing toxicity in the context of pollutant mineralization.