The Escherichia coli azoR gene promoter: A new sensing element for microbial biodetection of trace explosives

YiratHenshke¹, Benjamin Shemer¹, Shimshon Belkin

Department of Plant and Environmental Sciences, Institute of Life Sciences, The Hebrew University of Jerusalem, Israel.

Abstract

DNT (2,4-dinitrotoluene), a volatile impurity in military grade TNT-based explosives, is a potential tracer for the detection of buried landmines and other explosive devices. We have previously described DNT bioreporter strains, based on a fusion between the Escherichia coli yqjF gene promoter to either a bacterial luxCDABE luminescence gene cassette or to the green fluorescent protein gene gfp. We now present the azoR gene promoter, demonstrated to be strongly induced by DNT, as an alternative sensing element for this compound. An E. coli bioreporter strain harboring a plasmid-borne fusion of the azoR promoter region to Photorhabdus luminescens luxCDABE displayed stronger bioluminescent responses to DNT than the yqjF-based bioreporter. The azoR promoter was induced not directly by DNT but by its degradation products; the most potent inducer was 2,4,5-trihydroxytoluene (THT). This reaction was positively regulated by YhaJ, a member of the LysR family of transcriptional regulators, and was significantly amplified in a yhaK mutant. We also show that DNT degradation in E. coli involves glutathione, which plays a vital role in azoR gene promoter activation.

Keywords: azoR, Azoreductase, Dinitrotoluene, Explosives, Bioreporter, Landmines.