Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, USA.
Geobiology. 2010 Jan;8(1):56-68. doi: 10.1111/j.1472-4669.2009.00226.x. Epub 2009 Nov 25.
In an effort to improve the understanding of electron transfer mechanisms at the microbe-mineral interface, Shewanella oneidensis MR-1 mutants with in-frame deletions of outer-membrane cytochromes (OMCs), MtrC and OmcA, were characterized for the ability to reduce ferrihydrite (FH) using a suite of microscopic, spectroscopic, and biochemical techniques. Analysis of purified recombinant proteins demonstrated that both cytochromes undergo rapid electron exchange with FH in vitro with MtrC displaying faster transfer rates than OmcA. Immunomicroscopy with cytochrome-specific antibodies revealed that MtrC co-localizes with iron solids on the cell surface while OmcA exhibits a more diffuse distribution over the cell surface. After 3-day incubation of MR-1 with FH, pronounced reductive transformation mineral products were visible by electron microscopy. Upon further incubation, the predominant phases identified were ferrous phosphates including vivianite [Fe(3)(PO(4))(2)x8H(2)O] and a switzerite-like phase [Mn(3),Fe(3)(PO(4))(2)x7H(2)O] that were heavily colonized by MR-1 cells with surface-exposed outer-membrane cytochromes. In the absence of both MtrC and OmcA, the cells ability to reduce FH was significantly hindered and no mineral transformation products were detected. Collectively, these results highlight the importance of the outer-membrane cytochromes in the reductive transformation of FH and support a role for direct electron transfer from the OMCs at the cell surface to the mineral.
为了增进对微生物-矿物界面电子传递机制的理解,采用一系列微观、光谱和生化技术,对 Shewanella oneidensis MR-1 缺失外膜细胞色素(OMC)的突变体(MtrC 和 OmcA 基因的框内缺失)进行了特征描述,以评估其还原水铁矿(FH)的能力。对纯化的重组蛋白进行分析,结果表明两种细胞色素都能与 FH 在体外进行快速电子交换,其中 MtrC 的电子传递速率快于 OmcA。用细胞色素特异性抗体进行免疫显微镜分析表明,MtrC 与细胞表面的铁固体共定位,而 OmcA 在细胞表面的分布则更加弥散。在 MR-1 与 FH 孵育 3 天后,电子显微镜可见明显的还原转化矿物产物。进一步孵育后,鉴定出的主要相是亚铁磷酸盐,包括磷铁矿[Fe(3)(PO(4))(2)x8H(2)O]和类似于斯维泽矿的相[Mn(3),Fe(3)(PO(4))(2)x7H(2)O],这些相被大量定植在 MR-1 细胞表面暴露的外膜细胞色素上。当 MtrC 和 OmcA 都缺失时,细胞还原 FH 的能力受到显著阻碍,并且未检测到矿物转化产物。总的来说,这些结果强调了外膜细胞色素在 FH 还原转化中的重要性,并支持 OMC 直接从细胞表面向矿物传递电子的作用。
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