Tauriainen S, Karp M, Chang W, Virta M
Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland.
Appl Environ Microbiol. 1997 Nov;63(11):4456-61. doi: 10.1128/aem.63.11.4456-4461.1997.
Luminescent bacterial strains for the measurement of bioavailable arsenite and antimony were constructed. The expression of firefly luciferase was controlled by the regulatory unit of the ars operon of Staphylococcus aureus plasmid pI258 in recombinant plasmid pTOO21, with S. aureus RN4220, Bacillus subtilis BR151, and Escherichia coli MC1061 as host strains. Strain RN4220(pTOO21) was found to be the most sensitive for metal detection responding to arsenite, antimonite, and cadmium, the lowest detectable concentrations being 100, 33, and 330 nM, respectively. Strains BR151(pTOO21) and MC1061(pTOO21) responded to arsenite, arsenate, antimonite, and cadmium, the lowest detectable concentrations being 3.3 and 330 microM and 330 and 330 nM with BR151(pTOO21), respectively, and 3.3, 33, 3.3, and 33 microM with MC1061(pTOO21), respectively. In the absence of the mentioned ions, the expression of luciferase was repressed and only a small amount of background light was emitted. Other ions did not notably interfere with the measurement in any of the strains tested. Freeze-drying of the cells did not decrease the sensitivity of the detection of arsenite; however, the induction coefficients were somewhat lower.
构建了用于测定生物可利用亚砷酸盐和锑的发光细菌菌株。在重组质粒pTOO21中,萤火虫荧光素酶的表达由金黄色葡萄球菌质粒pI258的ars操纵子调控单元控制,以金黄色葡萄球菌RN4220、枯草芽孢杆菌BR151和大肠杆菌MC1061作为宿主菌株。发现菌株RN4220(pTOO21)对亚砷酸盐、亚锑酸盐和镉的金属检测最为敏感,最低可检测浓度分别为100 nM、33 nM和330 nM。菌株BR151(pTOO21)和MC1061(pTOO21)对亚砷酸盐、砷酸盐、亚锑酸盐和镉有反应,BR151(pTOO21)的最低可检测浓度分别为3.3 μM和330 nM以及330 nM和330 nM,而MC1061(pTOO21)的最低可检测浓度分别为3.3 μM、33 μM、3.3 μM和33 μM。在不存在上述离子的情况下,荧光素酶的表达受到抑制,仅发出少量背景光。其他离子在任何测试菌株中均未对测量产生显著干扰。细胞冻干并未降低亚砷酸盐检测的灵敏度;然而,诱导系数略低。