Cadmium-specific formation of metal sulfide ‘Q-particles’ by Klebsiella pneumoniae

Abstract

Summary: Klebsiella pneumoniae overcomes cadmium toxicity through the ‘biotransformation’ of cadmium ions into photoactive, nanometre-sized CdS particles deposited on the cell surface. The kinetics of particle formation during batch culture growth was monitored by electron microscopy (EM), energy-dispersive X-ray analysis and electronic absorption spectroscopy (EAS). During the deceleration phase of bacterial growth, the presence of CdS particles on the outer cell wall of K. pneumoniae (≥ 5 nm in diameter) was detected by EM. The size of these electron-dense particles continued to increase throughout the stationary phase of growth, with some of the particles reaching a diameter >200 nm. The formation of the extracellular CdS particles contributed to around 3-4% of the total cell biomass. EAS undertaken on these extracellular ‘bio-CdS’ particles suggested that the large ‘superparticles’ observed by EM, e.g. 200 nm, were aggregates of smaller particles termed ‘Q-particles’, ~ 4 nm in diameter. Metal sulfide particles were not formed in batch cultures of K. pneumoniae grown in the presence of lead, zinc, mercury, copper or silver ions. Growth in the presence of lead ions resulted in the formation of extracellular electron-dense particles containing lead but not sulfide or phosphate. Intracellular phosphorus-containing electron-opaque particles were formed during growth in the presence of copper and mercury. Intracellular electron-dense particles were formed in the presence of zinc ions but these did not contain phosphorus. From these results it was thought that metal sulfide formation in K. pneumoniae showed some cadmium-specificity. When cadmium and zinc ions were both added to the growth medium, metal sulfide particles were formed that were predominantly composed of cadmium, e.g. 48.6% cadmium and 0.04% zinc. Similarly, when cadmium and lead ions were both present during growth only CdS particles formed. In both cases analysis of the cells by EAS confirmed the presence of CdS only. These observations suggest that the mechanism of CdS formation is unlikely to occur simply through a cadmium-induced release of hydrogen sulfide by the cells into the external environment. If hydrogen sulfide production was the mechanism of sulfide formation then metal sulfide particles containing lead and zinc ions in addition to cadmium ions should have been produced.