Holmes Justin D, Richardson David J, Saed Shaheen, Evans-Gowing Richard, Russell David A, Sodeau John R
School of Chemical Sciences, University of East Anglia, Norwich NR4 7TJ, UK.
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK.
Microbiology (Reading). 1997 Aug;143 ( Pt 8):2521-2530. doi: 10.1099/00221287-143-8-2521.
Klebsiella pneumoniae overcomes cadmium toxicity through the 'biotrans-formation' 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 (> or = 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', approximately 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.
肺炎克雷伯菌通过将镉离子“生物转化”为沉积在细胞表面的光活性纳米级硫化镉颗粒来克服镉毒性。通过电子显微镜(EM)、能量色散X射线分析和电子吸收光谱(EAS)监测分批培养生长过程中颗粒形成的动力学。在细菌生长的减速期,通过EM检测到肺炎克雷伯菌外细胞壁上存在硫化镉颗粒(直径≥5nm)。在整个生长稳定期,这些电子致密颗粒的尺寸持续增加,一些颗粒直径超过200nm。细胞外硫化镉颗粒的形成约占细胞总生物量的3 - 4%。对这些细胞外“生物硫化镉”颗粒进行的EAS表明,EM观察到的大“超颗粒”,例如200nm的颗粒,是直径约4nm的较小颗粒“Q颗粒”的聚集体。在含有铅、锌、汞、铜或银离子的条件下培养的肺炎克雷伯菌分批培养物中未形成金属硫化物颗粒。在铅离子存在下生长导致形成含有铅但不含硫化物或磷酸盐的细胞外电子致密颗粒。在铜和汞存在下生长期间形成了细胞内含磷的电子不透明颗粒。在锌离子存在下形成了细胞内电子致密颗粒,但这些颗粒不含磷。从这些结果可以认为,肺炎克雷伯菌中金属硫化物的形成表现出一定的镉特异性。当将镉和锌离子都添加到生长培养基中时,形成的金属硫化物颗粒主要由镉组成,例如48.6%的镉和0.04%的锌。同样,当生长期间镉和铅离子都存在时,仅形成硫化镉颗粒。在这两种情况下,通过EAS对细胞进行分析均证实仅存在硫化镉。这些观察结果表明,硫化镉形成的机制不太可能仅仅是通过镉诱导细胞向外部环境释放硫化氢而发生。如果硫化氢的产生是硫化物形成的机制,那么除了镉离子之外还应该产生含有铅和锌离子的金属硫化物颗粒。