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Characterization of Thiomonas delicata arsenite oxidase expressed in Escherichia coli

机译:大肠杆菌中表达的Thiomonas delicata亚砷酸氧化酶的表征

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摘要

Microbial arsenite oxidation is an essential biogeochemical process whereby more toxic arsenite is oxidized to the less toxic arsenate. Thiomonas strains represent an important arsenite oxidizer found ubiquitous in acid mine drainage. In the present study, the arsenite oxidase gene (aioBA) was cloned from Thiomonas delicata DSM 16361, expressed heterologously in E. coli and purified to homogeneity. The purified recombinant Aio consisted of two subunits with the respective molecular weights of 91 and 21 kDa according to SDS-PAGE. Aio catalysis was optimum at pH 5.5 and 50–55 °C. Aio exhibited stability under acidic conditions (pH 2.5–6). The V max and K m values of the enzyme were found to be 4 µmol min−1 mg−1 and 14.2 µM, respectively. SDS and Triton X-100 were found to inhibit the enzyme activity. The homology model of Aio showed correlation with the acidophilic adaptation of the enzyme. This is the first characterization studies of Aio from a species belonging to the Thiomonas genus. The arsenite oxidase was found to be among the acid-tolerant Aio reported to date and has the potential to be used for biosensor and bioremediation applications in acidic environments.Electronic supplementary materialThe online version of this article (doi:10.1007/s13205-017-0740-7) contains supplementary material, which is available to authorized users.
机译:微生物亚砷酸盐的氧化是必不可少的生物地球化学过程,其中毒性更高的亚砷酸盐被氧化为毒性较小的砷酸盐。硫门氏菌菌株代表一种重要的砷氧化物,在酸性矿山排水中普遍存在。在本研究中,亚砷酸氧化酶基因(aioBA)是从Thiomonas delicata DSM 16361中克隆的,在大肠杆菌中异源表达并纯化至同质。纯化的重组Aio由两个亚基组成,根据SDS-PAGE分别具有91和21 kDa的分子量。在pH 5.5和50-55°C下,Aio催化效果最佳。 Aio在酸性条件下(pH 2.5-6)表现出稳定性。该酶的V max和K m值分别为4 µmol min -1 mg -1 和14.2 µM。发现SDS和Triton X-100抑制酶活性。 Aio的同源性模型显示出与该酶的嗜酸适应性相关。这是来自硫门氏菌属物种的Aio的首次表征研究。砷氧化酶是迄今为止报道的耐酸Aio之一,具有潜在的酸性环境中生物传感器和生物修复应用的潜力。电子补充材料本文的在线版本(doi:10.1007 / s13205-017-0740 -7)包含补充材料,授权用户可以使用。

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