Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University , Miami, Florida 33199, United States.
Environ Sci Technol. 2014 Jan 21;48(2):1141-7. doi: 10.1021/es4038319. Epub 2014 Jan 3.
The toxic metalloid arsenic is widely distributed in food, water, and soil. While inorganic arsenic enters the environment primarily from geochemical sources, methylarsenicals either result from microbial biotransformation of inorganic arsenic or are introduced anthropogenically. Methylarsenicals such as monosodium methylarsonic acid (MSMA) have been extensively utilized as herbicides, and aromatic arsenicals such as roxarsone (Rox) are used as growth promoters for poultry and swine. Organoarsenicals are degraded to inorganic arsenic. The toxicological effects of arsenicals depend on their oxidation state, chemical composition, and bioavailability. Here we report that the active forms are the trivalent arsenic-containing species. We constructed a whole-cell biosensor utilizing a modified ArsR repressor that is highly selective toward trivalent methyl and aromatic arsenicals, with essentially no response to inorganic arsenic. The biosensor was adapted for in vitro detection of organoarsenicals using fluorescence anisotropy of ArsR-DNA interactions. It detects bacterial biomethylation of inorganic arsenite both in vivo and in vitro with detection limits of 10(-7) M and linearity to 10(-6) M for phenylarsenite and 5 × 10(-6) M for methylarsenite. The biosensor detects reduced forms of MSMA and roxarsone and offers a practical, low cost method for detecting activate forms and breakdown products of organoarsenical herbicides and growth promoters.
有毒的类金属砷广泛存在于食物、水和土壤中。虽然无机砷主要从地球化学来源进入环境,但甲基砷化合物要么是由无机砷的微生物生物转化产生的,要么是人为引入的。甲基砷化合物如单甲基砷酸(MSMA)被广泛用作除草剂,而芳香砷化合物如洛克沙胂(Rox)则被用作家禽和猪的生长促进剂。有机砷会被降解为无机砷。砷化物的毒理学效应取决于其氧化态、化学组成和生物利用度。在这里,我们报告说,活性形式是含三价砷的物种。我们构建了一种全细胞生物传感器,利用一种经过改良的 ArsR 阻遏物,该阻遏物对三价甲基和芳香砷化合物具有高度选择性,对无机砷几乎没有反应。该生物传感器经过改造,可通过 ArsR-DNA 相互作用的荧光各向异性来检测有机砷化合物。它可以在体内和体外检测无机亚砷酸盐的细菌生物甲基化,对苯砷酸盐的检测限为 10(-7) M,线性度为 10(-6) M,对甲基砷酸盐的检测限为 5×10(-6) M。该生物传感器可检测 MSMA 和洛克沙胂的还原形式,并为检测有机砷除草剂和生长促进剂的活性形式和分解产物提供了一种实用且低成本的方法。