The Pseudomonas putida NfnB nitroreductase confers resistance to roxarsone.

The Pseudomonas putida NfnB nitroreductase confers resistance to roxarsone.
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DOI:
10.1016/j.scitotenv.2020.141339
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发表时间:
2020-12-15
期刊:
The Science of the total environment
影响因子:
--
通讯作者:
Rosen BP
Rosen BP
中科院分区:
其他
文献类型:
--
作者:
Chen J;Rosen BP

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Roxarsone(3-硝基-4-羟基苯胂酸,Rox)已被用作家禽和猪的抗菌生长促进剂数十年。洛克沙胂以不变的形式在鸡粪中排泄,可通过微生物转化为各种含砷化合物,如3-氨基-4-羟基苯胂酸(HAPA(V)),污染环境并对健康构成潜在危害。为了科普砷的毒性,几乎每一种原核生物都有一个ars(抗砷)操纵子,其中一些赋予对洛克沙胂的抗性。恶臭假单胞菌KT 2440是一种能够代谢多种芳香族化合物的环境菌株,被用作芳香族化合物生物降解的模式生物。在这里,我们报告说,恶臭假单胞菌KT 2440(ΔΔars),其中两个ars操纵子已被删除,保留对剧毒的三价Rox(III),洛克沙胂的可能的活性形式的抗性。本研究利用构建的恶臭假单胞菌KT 2440(ΔΔars)基因组文库筛选大肠杆菌对Rox(III)的抗性。一个基因,命名为,PpnfnB,被确定为编码一个假定的6,7-二氢蝶啶还原酶。表达PpnfnB的细胞还原Rox(III)的硝基,并且纯化的NfnB催化Rox(III)的FMN-NADPH依赖性硝基还原成毒性较小的HAPA(III)。这确定了合成芳香族砷化合物分解的关键步骤。
Roxarsone (3-nitro-4-hydroxyphenylarsonic acid, Rox) has been used for decades as an antimicrobial growth promoter for poultry and swine. Roxarsone is excreted in chicken manure unchanged and can be microbially transformed into a variety of arsenic-containing compounds such as 3-amino-4-hydroxyphenylarsonic acid (HAPA(V)) that contaminate the environment and present a potential health hazard. To cope with arsenic toxicity, nearly every prokaryote has an ars (arsenic resistance) operon, some of which confer resistance to roxarsone. Pseudomonas putida KT2440 is a robust environmental isolate capable of metabolizing many aromatic compounds and is used as a model organism for biodegradation of aromatic compounds. Here we report that P. putida KT2440 (ΔΔars) in which the two ars operons had been deleted retains resistance to highly toxic trivalent Rox(III), the likely active form of roxarsone. In this study, a genomic library constructed from P. putida KT2440 (ΔΔars) was used to screen for resistance to Rox(III) in Escherichia coli. One gene, termed, PpnfnB, was identified that encodes a putative 6,7-dihydropteridine reductase. Cell expressing PpnfnB reduce the nitro group of Rox(III), and purified NfnB catalyzes FMN-NADPH-dependent nitroreduction of Rox(III) to less toxic HAPA(III). This identifies a key step in the breakdown of synthetic aromatic arsenicals.
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