Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
Institute of Environment Remediation and Human Health, and College of Ecology and Environment, Southwest Forestry University, Kunming, 650224, China.
Environ Microbiol. 2021 Dec;23(12):7550-7562. doi: 10.1111/1462-2920.15817. Epub 2021 Oct 21.
Toxic organoarsenicals enter the environment from biogenic and anthropogenic activities such as microbial methylation of inorganic arsenic and pentavalent herbicides such as monosodium methylarsenate (MSMA or MAs(V)). Trivalent MAs(III) is considerably more toxic than arsenite or arsenate. Microbes have evolved mechanisms to detoxify organoarsenicals. We previously identified ArsV, a flavin-linked monooxygenase and demonstrated that it confers resistance to methylarsenite by oxidation to methylarsenate. The arsV gene is usually in an arsenic resistance (ars) operon controlled by an ArsR repressor and adjacent to a methylarsenite efflux gene, either arsK or a gene for a putative transporter. Here we show that Paracoccus sp. SY oxidizes methylarsenite. It has an ars operon with three genes, arsR, arsV and a transport gene termed arsW. Heterologous expression of arsV in Escherichia coli conferred resistance to MAs(III), while arsW did not. Co-expression of arsV and arsW increased resistance compared with either alone. The cells oxidized methylarsenite and accumulated less methylarsenate. Everted membrane vesicles from E. coli cells expressing arsW-accumulated methylarsenate. We propose that ArsV is a monooxygenase that oxidizes methylarsenite to methylarsenate, which is extruded by ArsW, one of only a few known pentavalent organoarsenical efflux permeases, a novel pathway of organoarsenical resistance.
有毒的有机胂化合物通过生物和人为活动进入环境,例如无机砷的微生物甲基化和五价除草剂,如单甲基砷酸钠(MSMA 或 MAs(V))。三价 MAs(III) 的毒性比亚砷酸盐或砷酸盐大得多。微生物已经进化出了解毒有机胂化合物的机制。我们之前鉴定出 ArsV,这是一种黄素连接的单加氧酶,并证明它通过氧化为甲基砷酸盐来赋予对甲基砷酸盐的抗性。arsV 基因通常位于砷抗性(ars)操纵子中,受 ArsR 抑制剂控制,并且与甲基砷酸盐外排基因相邻,该基因是 arsK 或假定转运蛋白的基因。在这里,我们表明 Paracoccus sp. SY 氧化甲基砷酸盐。它有一个 Ars 操纵子,其中包含三个基因,arsR、arsV 和一个称为 arsW 的转运基因。arsV 在大肠杆菌中的异源表达赋予了对 MAs(III)的抗性,而 arsW 则没有。arsV 和 arsW 的共表达比单独表达时增加了抗性。细胞氧化甲基砷酸盐并积累较少的甲基砷酸盐。表达 arsW 的大肠杆菌细胞的外翻膜囊泡积累了甲基砷酸盐。我们提出 ArsV 是一种单加氧酶,它将甲基砷酸盐氧化为甲基砷酸盐,而 ArsW 将其排出,ArsW 是少数已知的五价有机胂外排转运蛋白之一,这是一种有机胂抗性的新途径。