Yoon Youngdae, Kim Sunghoon, Chae Yooeun, Kim Shin Woong, Kang Yerin, An Gyeonghyeon, Jeong Seung-Woo, An Youn-Joo
Department of Environmental Health Science, Konkuk University, Seoul, 05029, South Korea.
Department of Environmental Engineering, Kunsan National University, Kunsan, 54150, South Korea.
Appl Microbiol Biotechnol. 2016 Apr;100(8):3713-22. doi: 10.1007/s00253-016-7338-6. Epub 2016 Feb 6.
Whole-cell bioreporters (WCBs) have attracted increasing attention during the last few decades because they allow fast determination of bioavailable heavy metals in contaminated sites. Various WCBs to monitor specific heavy metals such as arsenic and cadmium in diverse environmental systems are available. However, currently, no study on simultaneous analysis of arsenic and cadmium has been reported, even though soils are contaminated by diverse heavy metals and metalloids. We demonstrated herein the development of dual-sensing WCBs to simultaneously quantify bioavailable arsenic and cadmium in contaminated sites by employing the promoter regions of the ars and znt operons as separate metal-sensing domains, and egfp and mcherry as reporter genes. The dual-sensing WCBs were generated by inserting two sets of genes into E. coli DH5α. The capability of WCBs was successfully proved to simultaneously quantify bioavailable arsenic and cadmium in amended Landwirtschaftliche Untersuchungs und Forschungsanstalt (LUFA) soils, and then, it was applied to contaminated field soils collected from a smelter area in Korea. As a result, it was noticed that the bioavailable portion of cadmium was higher than that of arsenic while the absolute amount of bioavailable arsenic and cadmium level was opposite. Since both cadmium and arsenic were assessed from the same E. coli cells, the data obtained by using dual-sensing WCBs would be more efficient and convenient than that from comparative WCB assay. In spite of advantageous aspects, to our knowledge, this is the first report on a dual-sensing WCB for rapid and concurrent quantification of bioavailable arsenic and cadmium in contaminated soils.
在过去几十年中,全细胞生物传感器(WCBs)受到了越来越多的关注,因为它们能够快速测定污染场地中生物可利用的重金属。目前已有各种用于监测不同环境系统中特定重金属(如砷和镉)的WCBs。然而,尽管土壤受到多种重金属和类金属的污染,但目前尚未有关于同时分析砷和镉的研究报道。在本文中,我们展示了双感应WCBs的开发,通过利用ars和znt操纵子的启动子区域作为单独的金属感应域,以及egfp和mcherry作为报告基因,来同时定量污染场地中生物可利用的砷和镉。通过将两组基因插入大肠杆菌DH5α中来构建双感应WCBs。成功证明了WCBs能够同时定量改良的德国农业研究与试验站(LUFA)土壤中生物可利用的砷和镉,然后将其应用于从韩国一个冶炼厂区域采集的污染场地土壤。结果发现,镉的生物可利用部分高于砷,而生物可利用砷和镉的绝对含量则相反。由于镉和砷都是从同一大肠杆菌细胞中评估的,因此使用双感应WCBs获得的数据将比比较WCB分析的数据更高效、更方便。尽管有这些优势,但据我们所知,这是第一份关于用于快速同时定量污染土壤中生物可利用砷和镉的双感应WCB的报告。