Multiple mechanisms collectively mediate tungsten homeostasis and resistance in Citrobacter sp. Lzp2

Tungsten (W) is an emerging contaminant, and current knowledge on W resistance profiles of microorganisms remains scarce and fragmentary. This study aimed to explore the physiological responses of bacteria under W stress and to resolve genes and metabolic pathways involved in W resistance using a transcriptome expression profiling assay. The results showed that the bacterium Citrobacter sp. Lzp2, screened from W-contaminated soil, could tolerate hundreds of mM W(VI) with a 50% inhibiting concentration of ∼110 mM. To cope with W stress, Citrobacter sp. Lzp2 secreted large amounts of proteins through the type VI secretory system (T6SS) to chelate W oxoanions via carboxylic groups in extracellular polymeric substances (EPS), and could transport cytosolic W outside via the multidrug efflux pumps (mdtABC and acrD). Intracellular W is probably bound by chaperone proteins and metal-binding pterin (tungstopterin) through the sulfur relay system. We propose that tetrathionate respiration is a new metabolic pathway for cellular W detoxification likely producing thio-tungstate. We conclude that multiple mechanisms collectively mediate W homeostasis and resistance in Citrobacter sp. Lzp2. Our results have important implications not only for understanding the intricate regulatory network of W homeostasis in microbes but also for bio-recovery and bioremediation of W in contaminated environments.