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具有高砷挥发能力的菌株分离及甲基转移酶基因鉴定

Isolation of a strain and identification of methyltransferase genes conferring the high arsenic volatilizing ability.

作者信息

Singh Diksha, Sharma Nitish, Agarwal Sheetal, Khan Sadaf Aiman, Jain Veena, Singh Sukhveer, Roy Somendu, Yadav Kusum, Singh Sudhir Pratap, Srivastava Vikas

机构信息

Systems Toxicology Group, FEST Division, CSIR - Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India.

Department of Biochemistry, University of Lucknow, Lucknow, Uttar Pradesh, India.

出版信息

Appl Environ Microbiol. 2025 Jun 18;91(6):e0246724. doi: 10.1128/aem.02467-24. Epub 2025 May 30.

Abstract

Microbial arsenic (As) volatilization plays a significant role in the global As biogeochemical cycle and presents a promising approach for bioremediation. In this study, an autotrophic strain of was isolated from sewage water containing 75 ppb of arsenic, exhibiting a high arsenic metabolism and volatilization rate. This strain tolerated arsenic concentrations ranging from 10 to 500 ppm. Volatilization tests were conducted at an environmental median of 200 ppb (0.2 ppm) As. The strain eliminated 50% of the total arsenic within 48 h, and 0.1 ppm (101 ppb) of arsenic was reported to be trapped in the headspace of the culture tube, confirming its volatilization. After genome isolation, sequencing, and assembly, the existence of the As metabolizing operon (RABC), as well as other As-resistance conferring genes (M, I, A, B, and A), was identified using different annotation methodologies. The genes conferring resistance against different heavy metals and multiple drugs used as antimicrobials were also identified in the genome assembly of the isolate. Based on the gene expression measured using real-time polymerase chain reaction, complete gene sequences of four M genes were mined, cloned, and expressed in BL21(DE3) cells. The cells harboring recombinant plasmid of each construct could volatilize As up to 86 ppb after 48 h incubation in arsenic-enriched media. The isolated strain of and its heterologously expressed methyltransferase genes could be a potent tool for permanently removing arsenic from water samples by volatilization.IMPORTANCEArsenic contamination in water, soil, and air poses significant health and environmental risks, as inorganic arsenic compounds are highly toxic and carcinogenic. Microorganisms capable of transforming arsenic into volatile forms play a pivotal role in the biogeochemical cycling of this metalloid, reducing its bioavailability and toxicity in contaminated environments. In this work, a strain of sp. was isolated from the sewage water and tested for its ability to survive in minimal arsenic media. The strain was found to be highly resistant to arsenic and volatilized more than 50% of the arsenic from the growth media. The putative methyltransferase genes from the isolated strain, when heterologously expressed in , conferred an ability to volatilize arsenic in the recombinant host, too. Therefore, the isolated microorganism offers a natural, eco-friendly alternative to conventional chemical methods, making it an important tool for addressing arsenic biosafety issues in the environment.

摘要

微生物砷(As)挥发在全球砷生物地球化学循环中起着重要作用,是一种很有前景的生物修复方法。在本研究中,从含75 ppb砷的污水中分离出一株自养菌株,该菌株表现出高砷代谢和挥发率。该菌株耐受10至500 ppm的砷浓度。挥发试验在环境中值200 ppb(0.2 ppm)的砷浓度下进行。该菌株在48小时内去除了总砷的50%,据报道培养管顶空中捕获了0.1 ppm(101 ppb)的砷,证实了其挥发作用。在进行基因组分离、测序和组装后,使用不同的注释方法鉴定了砷代谢操纵子(RABC)以及其他赋予砷抗性的基因(M、I、A、B和A)的存在。在该分离株的基因组组装中还鉴定出了赋予对不同重金属和多种用作抗菌剂的药物抗性的基因。基于使用实时聚合酶链反应测量的基因表达,挖掘、克隆了四个M基因的完整基因序列,并在BL21(DE3)细胞中进行了表达。在富含砷的培养基中培养48小时后,携带每个构建体重组质粒的细胞能够将砷挥发至86 ppb。分离出的菌株及其异源表达的甲基转移酶基因可能是通过挥发从水样中永久去除砷的有效工具。

重要性

水、土壤和空气中的砷污染带来重大的健康和环境风险,因为无机砷化合物具有高毒性和致癌性。能够将砷转化为挥发性形式的微生物在这种类金属的生物地球化学循环中起着关键作用,降低了其在受污染环境中的生物可利用性和毒性。在这项工作中,从污水中分离出一株[具体菌株名称]菌株,并测试了其在最低砷培养基中的存活能力。发现该菌株对砷具有高度抗性,并且从生长培养基中挥发了超过50%的砷。从分离菌株中获得的假定甲基转移酶基因在[具体宿主名称]中异源表达时,也赋予了重组宿主挥发砷的能力。因此,分离出的微生物为传统化学方法提供了一种天然、环保的替代方法,使其成为解决环境中砷生物安全问题的重要工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a45/12175517/6374453fb040/aem.02467-24.f001.jpg

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