Profiling of the reactive oxygen species-related ecotoxicity of CuO, ZnO, TiO2, silver and fullerene nanoparticles using a set of recombinant luminescent Escherichia coli strains: differentiating the impact of particles and solubilised metals
Profiling of the reactive oxygen species-related ecotoxicity of CuO, ZnO, TiO2, silver and fullerene nanoparticles using a set of recombinant luminescent Escherichia coli strains: differentiating the impact of particles and solubilised metals
复制标题
DOI：
10.1007/s00216-010-3962-7
复制
发表时间：
2010-09-01
影响因子：
4.3
通讯作者：
Kahru, A.
中科院分区：
文献类型：
--
作者：
Ivask, A.;Bondarenko, O.;Kahru, A.
我们提出了一种新的高通量发光细菌测试的组合，用于评估工程纳米颗粒（eNPs）的活性氧（ROS）生成潜力和溶解的金属离子在这一过程中的作用。本试验包括不同工程改造的重组大肠杆菌菌株：（1）一种新的传感器菌株，其生物发光由超氧阴离子诱导;（2）6个重组大肠杆菌菌株。大肠杆菌菌株（超氧化物歧化酶（sod）单突变体、双突变体和三突变体以及相应的野生型菌株），其用luxCDABE基因转化，所述基因通过降低毒性化合物的发光来响应毒性化合物;以及（3）其中生物发光被生物可利用的金属（Cu、Zn和Ag）特异性诱导的三种菌株。在nTiO（2）、nCuO、nZnO和nAg（分别为25、30、70和< 100 nm）NP和富勒烯上评估了该测试组在分析eNP的氧化电位中的适用性。作为尺寸或溶解度的对照，还分析了本体制剂（bTiO（2）、bCuO和bZnO）和可溶性盐（ZnSO 4、CuSO 4和AgNO 3）。细菌毒性试验表明，nCuO对三重SOD突变体的毒性是野生型的4倍，nAg对三重SOD突变体的毒性是野生型的15倍（2 h EC_（50）值分别为8.1和2.0mgCu_l（-1），46和3.1mgAg_l（-1））。超氧阴离子诱导菌株证实了nCuO和nAg形成ROS。金属传感器细菌表明，CuO NPs的ROS形成是由溶解的Cu离子引起的，但在nAg的情况下，颗粒也有影响。nZnO对sod三突变株的毒性显著高于野生型（2 h EC_（50）分别为4.5和54 mg Zn·l ~（-1））。富勒烯在3，882 mg l（-1）时抑制sod三突变体的生物发光，但即使在20，800 mg l（-1）时对野生型菌株也没有影响。纳米和bTiO（2）仅在高浓度（> 4，000 mg l（-1））下对细菌的存活率显示出一定的影响，尽管nTiO（2）（而不是bTiO（2））从100 mg l（-1）开始诱导超氧阴离子敏感细菌的生物发光。因此，我们的创新组合方法有望提供关于一般毒性、ROS产生潜力以及金属纳米颗粒情况下金属溶解的更一致和信息丰富的数据。
We propose a novel combination of high-throughput luminescent bacterial tests for the evaluation of the reactive oxygen species (ROS)-generating potential of engineered nanoparticles (eNPs) and the role of solubilised metal ions in this process. The set of tests consists of differently engineered recombinant Escherichia coli strains: (1) a new sensor strain, which bioluminescence is induced by superoxide anions; (2) six recombinant E. coli strains (superoxide dismutase (sod) single, double and triple mutants and a respective wild-type strain), transformed with luxCDABE genes responding to toxic compounds by decreasing their luminescence; and (3) three strains in which bioluminescence is specifically induced by bioavailable metals (Cu, Zn and Ag). The applicability of this battery of tests in profiling oxidative potential of eNPs was evaluated on nTiO(2), nCuO, nZnO and nAg (25, 30, 70 and < 100 nm, respectively) NPs and fullerenes. As controls for the size or solubility, the bulk formulations (bTiO(2), bCuO and bZnO) and soluble salts (ZnSO4, CuSO4 and AgNO3) were also analysed. Bacterial toxicity tests showed that nCuO was four-fold more toxic, and nAg was 15-fold more toxic to triple sod mutant than to wild type (2-h EC50 values were 8.1 and 2.0 mg Cu l(-1), respectively, and 46 and 3.1 mg Ag l(-1), respectively). Formation of ROS by nCuO and nAg was proved by superoxide anion-inducible strain. The metal sensor bacteria showed that the ROS formation by CuO NPs was caused by solubilised Cu ions, but in case of nAg, particles also had an effect. nZnO was remarkably more toxic to sod triple mutant than to wild type strain (2-h EC50 were 4.5 and 54 mg Zn l(-1), respectively). Fullerenes inhibited the bioluminescence of sod triple mutant at 3,882 mg l(-1) but had no effect on the wild-type strain even at 20,800 mg l(-1). Nano and bTiO(2) showed some effect on viability of bacteria only at high concentrations (> 4,000 mg l(-1)) although nTiO(2) (but not bTiO(2)) induced the bioluminescence of the superoxide anion sensing bacteria starting from 100 mg l(-1). Thus, our innovative combined approach is expected to provide more consistent and informative data concerning the general toxicity, ROS-production potential and also solubilisation of metals in the case of metallic NPs.