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Stenotrophomonas maltophilia SeITE02 a New Bacterial Strain Suitable for Bioremediation of Selenite-Contaminated Environmental Matrices

机译:嗜麦芽窄食单胞菌SeITE02一种新型细菌菌株适用于亚硒酸盐污染的环境基质的生物修复。

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摘要

Biochemical and proteomic tools have been utilized for investigating the mechanism of action of a new Stenotrophomonas maltophilia strain (SeITE02), a gammaproteobacterium capable of resistance to high concentrations of selenite [SeO32−, Se(IV)], reducing it to nontoxic elemental selenium under aerobic conditions; this strain was previously isolated from a selenite-contaminated mining soil. Biochemical analysis demonstrated that (i) nitrite reductase does not seem to take part in the process of selenite reduction by the bacterial strain SeITE02, although its involvement in this process had been hypothesized in other cases; (ii) nitrite strongly interferes with selenite removal when the two oxyanions (NO2 and SeO32−) are simultaneously present, suggesting that the two reduction/detoxification pathways share a common enzymatic step, probably at the level of cellular transport; (iii) in vitro, selenite reduction does not take place in the membrane or periplasmic fractions but only in the cytoplasm, where maximum activity is exhibited at pH 6.0 in the presence of NADPH; and (iv) glutathione is involved in the selenite reduction mechanism, since inhibition of its synthesis leads to a considerable delay in the onset of reduction. As far as the proteomic findings are concerned, the evidence was reached that 0.2 mM selenite and 16 mM nitrite, when added to the culture medium, caused a significant modulation (ca. 10%, i.e., 96 and 85 protein zones, respectively) of the total proteins visualized in the respective two-dimensional maps. These spots were identified by mass spectrometry analysis and were found to belong to the following functional classes: nucleotide synthesis and metabolism, damaged-protein catabolism, protein and amino acid metabolism, and carbohydrate metabolism along with DNA-related proteins and proteins involved in cell division, oxidative stress, and cell wall synthesis.
机译:已经使用生化和蛋白质组学工具研究了新的嗜麦芽嗜单胞菌麦芽嗜热菌株(SeITE02)的作用机理,该菌株能够抵抗高浓度的亚硒酸盐[SeO3 2-,Se(IV)] ,在有氧条件下将其还原为无毒元素硒;该菌株以前是从亚硒酸盐污染的采矿土壤中分离出来的。生化分析表明:(i)亚硝酸还原酶似乎不参与细菌菌株SeITE02的亚硒酸盐还原过程,尽管在其他情况下也推测其参与了该过程; (ii)当同时存在两个氧阴离子(NO2 -和SeO3 2-)时,亚硝酸盐强烈干扰亚硒酸盐的去除,表明这两种还原/解毒途径具有相同的酶促步骤,可能在细胞运输水平; (iii)在体外,亚硒酸盐的还原不会发生在膜或周质部分,而只会发生在细胞质中,在存在NADPH的情况下,在pH值为6.0时具有最大的活性; (iv)谷胱甘肽参与亚硒酸盐的还原机理,因为对其合成的抑制导致还原开始的相当大的延迟。就蛋白质组学研究结果而言,已获得证据表明,将0.2 mM亚硒酸盐和16 mM亚硝酸盐添加到培养基中后,会显着调节蛋白质的表达(分别约为10%,即96和85个蛋白区)。在相应的二维图中可视化的总蛋白质。这些斑点通过质谱分析鉴定,并且被发现属于以下功能类别:核苷酸合成和代谢,受损的蛋白质分解代谢,蛋白质和氨基酸代谢,碳水化合物代谢以及与DNA相关的蛋白质和参与细胞分裂的蛋白质,氧化应激和细胞壁合成。

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