Identification of a Novel Chromate and Selenite Reductase FesR in Alishewanella sp. WH16-1.
Identification of a Novel Chromate and Selenite Reductase FesR in Alishewanella sp. WH16-1.
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DOI：
10.3389/fmicb.2022.834293
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发表时间：
2022
影响因子：
5.2
通讯作者：
Xia X
中科院分区：
文献类型：
--
作者：
Zhou Z;Zhu L;Dong Y;You L;Zheng S;Wang G;Xia X
铁氧还蛋白（AAY72_06850，命名为FesR）被鉴定为与Alishewanella sp. WH 16 -1中的铬酸盐[Cr（VI）]抗性相关。FesR及其类似蛋白质与其他还原酶家族在遗传学上是分离的。与已报道的Cr（VI）和Se（IV）还原酶不同，在FesR序列中发现了两个4Fe-4S簇和一个黄素腺嘌呤二核苷酸（FAD）结合结构域。体内实验表明，突变株ΔfesR与野生型和互补株相比，丧失了部分Cr（VI）和Se（IV）的还原能力。此外，在大肠杆菌中的过表达和体外酶试验表明，FesR参与Cr（VI）和Se（IV）的还原。用紫外-可见光谱（UV-VIS）和电子顺磁共振（EPR）检测了纯化后的FesR中的4Fe-4S簇合物。FesR还原Cr（VI）和Se（IV）的Km值分别为1682.0 ± 126.2和1164.0 ± 89.4 μmol/L，Vmax值分别为4.1 ± 0.1和9.4 ± 0.3 μmol min-1 mg-1。此外，定点突变和氧化还原电位分析表明，4Fe-4S团簇是FesR所必需的，FAD可以提高FesR作为细胞内电子转运体的酶效率。据我们所知，FesR是一种新的Cr（VI）和Se（IV）还原酶。
A ferredoxin protein (AAY72_06850, named FesR) was identified to associate with chromate [Cr(VI)] resistance in Alishewanella sp. WH16-1. FesR and its similar proteins were phylogenetically separated from other reductase families. Unlike the reported Cr(VI) and selenite [Se(IV)] reductases, two 4Fe-4S clusters and one flavin adenine dinucleotide (FAD) -binding domain were found in the FesR sequence. The experiment in vivo showed that the mutant strain ΔfesR had lost partial Cr(VI) and Se(IV) reduction capacities compared to the wild-type and complemented strains. Furthermore, overexpression in Escherichia coli and enzymatic tests in vitro showed FesR were involved in Cr(VI) and Se(IV) reduction. 4Fe-4S cluster in purified FesR was detected by ultraviolet-visible spectrum (UV-VIS) and Electron Paramagnetic Resonance (EPR). The Km values of FesR for Cr(VI) and Se(IV) reduction were 1682.0 ± 126.2 and 1164.0 ± 89.4 μmol/L, and the Vmax values for Cr(VI) and Se(IV) reduction were 4.1 ± 0.1 and 9.4 ± 0.3 μmol min–1 mg–1, respectively. Additionally, site-directed mutagenesis and redox potential analyses showed that 4Fe-4S clusters were essential to FesR, and FAD could enhance the enzyme efficiencies of FesR as intracellular electron transporters. To the best of our knowledge, FesR is a novel Cr(VI) and Se(IV) reductase.
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影响因子：
4.4
作者：
Ackerley, DF;Gonzalez, CF;Matin, A
通讯作者：
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DOI：
10.1080/21691401.2016.1228660
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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