No abstract

Previous work documented the general antibacterial mechanism of iron containing clays that involved hydroxyl radical (•OH) production from soluble Fe, and attack of cell membrane and intracellular proteins. Here we explore the role of clay structural Fe(II) in •OH production at near neutral pH and identify a lipid involved in the antibacterial process. Structural Fe(III) in nontronite NAu-2 was reduced (rNAu-2) and E. coli, a model bacterium, was exposed to rNAu-2 in oxic suspension. The antibacterial activity of rNAu-2 was dependent on pH and Fe(II) concentration, where E. coli were completely killed at pH 6, but survived at pH 7 and 8. In the presence of a •OH scavenger or in anaerobic atmosphere, E. coli survived better, suggesting that cell death may be caused by •OH generated from oxidation of structural Fe(II) in rNAu-2. In-situ imaging revealed damage of a membrane lipid, cardiolipin, in the polar region of E. coli cells, where reactive oxygen species and redox-active labile Fe were enriched. Our results advance the previous antibacterial model by demonstrating that the structural Fe(II) is the primary source of •OH, which damages cardiolipin, triggers the influx of soluble Fe into the cell, and ultimately leads to cell death.

Previous studies have documented the antibacterial activity of certain iron-containing clays. However, the repulsion between negatively charged bacteria and the clay surface makes this process inefficient. The objective of this study is to improve the bactericidal efficiency of clays by reversing their surface charge from negative to positive. To achieve this objective, positively charged chitosan, a nontoxic and biodegradable polymer, was intercalated into nontronite NAu-2. Chitosan-intercalated NAu-2 (C-NAu-2) was chemically reduced to obtain reduced C-NAu-2 (rC-NAu-2). Relative to reduced nontronite (rNAu-2), the antibacterial activity of rC-NAu-2 is higher and more persistent over a pH range of 6–8. The close spatial association between positively charged rC-NAu-2 and negatively charged bacteria increases the chances of cell membrane attack by extracellular ROS, the influx of soluble Fe2+ into the bacterial cell, and the yield of intracellular ROS. All these factors contribute to the enhanced antibacterial activity of rC-NAu-2. In contrast to rNAu-2 treated E. coli cells, where membrane damage and intracellular ROS/Fe accumulation are restricted to the polar regions, the close bacteria–clay association in rC-NAu-2 results in nonselective membrane damage and more uniform intracellular ROS/Fe distribution across whole bacterial cells. These results advance the antibacterial model by highlighting the importance of bacteria–clay interactions to the antibacterial activity of Fe-bearing clays.

Reduction of U(VI) to U(IV) drastically reduces its solubility and has been proposed as a method for remediation of uranium contamination. However, much is still unknown about the kinetics, mechanisms, and products of U(VI) bioreduction in complex systems. In this study, U(VI) bioreduction experiments were conducted with Shewanella putrefaciens strain CN32 in the presence of clay minerals and two organic ligands: citrate and EDTA. In reactors with U and Fe(III)−clay minerals, the rate of U(VI) bioreduction was enhanced due to the presence of ligands, likely because soluble Fe 3+ − and Fe 2+ −ligand complexes served as electron shuttles. In the presence of citrate, bioreduced U(IV) formed a soluble U(IV)−citrate complex in experiments with either Fe-rich or Fe-poor clay mineral. In the presence of EDTA, U(IV) occurred as a soluble U(IV)−EDTA complex in Fe-poor montmorillonite experiments. However, U(IV) remained associated with the solid phase in Fe-rich nontronite experiments through the formation of a ternary U(IV)−EDTA−surface complex, as suggested by the EXAFS analysis. Our study indicates that organic ligands and Fe(III)-bearing clays can significantly affect the microbial reduction of U(VI) and the stability of the resulting U(IV) phase.

Reduced nontronite has been demonstrated to be antibacterial through the production of hydroxyl radical (•OH) from the oxidation of structural Fe­(II). Herein, we investigated the antibacterial activity of more common smectite–illite (S–I) clays toward Escherichia coli cells, including montmorillonite SWy-3, illite IMt-2, 50–50 S–I rectorite RAr-1, 30–70 S–I ISCz-1, and nontronite NAu-2. Under an oxic condition, reduced clays (with a prefix r before mineral names) produced reactive oxygen species (ROS), and the antibacterial activity followed the order of rRAr-1 > rSWy-3 ≥ rNAu-2 ≫ rIMt-2 ≥ rISCz-1. The strongest antibacterial activity of rRAr-1 was contributed by a combination of •OH and Fe­(IV) generated from structural Fe­(II)/adsorbed Fe2+ and soluble Fe2+, respectively. Higher levels of lipid and protein oxidation, intracellular ROS accumulation, and membrane disruption were consistent with this antibacterial mechanism of rRAr-1. The antibacterial activity of other S–I clays depended on layer expandability, which determined the reactivity of structural Fe­(II) and the production of •OH, with the expandable smectite being the most antibacterial and nonexpandable illite the least. Our results provide new insights into the antibacterial mechanisms of clay minerals.