Department of Bioengineering, School of Minerals Processing and Bioengineering, Central South University, 410083, Changsha, Hunan Province, China.
Folia Microbiol (Praha). 2013 Nov;58(6):551-60. doi: 10.1007/s12223-013-0244-8. Epub 2013 Apr 7.
Metal-related genes (afe_0654, afe_0671, afe_0674, afe_1143, afe_1144, and afe_2126) were cloned to identify whether those genes existed in Acidithiobacillus ferrooxidans strain DC (A. ferrooxidans DC). The deduced amino acid sequences of those genes were analyzed by bioinformatics. The tolerance levels of A. ferrooxidans DC to Mn(2+), Zn(2+), and Cd(2+) were determined, which were 0.52, 0.42, and 0.16 mol/L for ferrous iron-grown cells and 0.38, 0.18, and 0.08 mol/L for sulfur-grown cells, respectively. Real-time quantitative PCR was employed to analyze the transcriptional levels of the metal-related genes when ferrous iron- and sulfur-grown cells of A. ferrooxidans DC, respectively, exposed to Mn(2+), Zn(2+), and Cd(2+). The metal-related genes were up-regulated when A. ferrooxidans DC exposed to Mn(2+). When A. ferrooxidans DC exposed to Zn(2+), the metal-related genes were up-regulated in sulfur-grown cells; afe_0654 and afe_0674 were down-regulated, and the others were up-regulated in ferrous iron-grown cells. Afe_2126 was down-regulated, and the others were up-regulated when A. ferrooxidans DC exposed to Cd(2+). According to experimental results and bioinformatics analysis, the proteins encoded by afe_0654 and afe_0674 may relate with Mn(2+) and Cd(2+) efflux. It needed further study whether they relate with Zn(2+) transport. Proteins encoded by afe_0671, afe_1143, and afe_1144 may relate with the efflux of Mn(2+), Zn(2+), and Cd(2+). The protein encoded by afe_2126 may relate with Mn(2+) and Zn(2+) efflux and Cd(2+) uptake.
金属相关基因(afe_0654、afe_0671、afe_0674、afe_1143、afe_1144 和 afe_2126)被克隆以鉴定这些基因是否存在于嗜酸氧化亚铁硫杆菌菌株 DC(A. ferrooxidans DC)中。通过生物信息学分析这些基因的推导氨基酸序列。确定了 A. ferrooxidans DC 对 Mn(2+)、Zn(2+)和 Cd(2+)的耐受水平,亚铁铁生长细胞分别为 0.52、0.42 和 0.16 mol/L,硫生长细胞分别为 0.38、0.18 和 0.08 mol/L。实时定量 PCR 用于分析 A. ferrooxidans DC 的亚铁铁和硫生长细胞分别暴露于 Mn(2+)、Zn(2+)和 Cd(2+)时金属相关基因的转录水平。当 A. ferrooxidans DC 暴露于 Mn(2+)时,金属相关基因上调。当 A. ferrooxidans DC 暴露于 Zn(2+)时,硫生长细胞中金属相关基因上调;亚铁铁生长细胞中 afe_0654 和 afe_0674 下调,其余基因上调。当 A. ferrooxidans DC 暴露于 Cd(2+)时,afe_2126 下调,其余基因上调。根据实验结果和生物信息学分析,afe_0654 和 afe_0674 编码的蛋白质可能与 Mn(2+)和 Cd(2+)外排有关。需要进一步研究它们是否与 Zn(2+)运输有关。afe_0671、afe_1143 和 afe_1144 编码的蛋白质可能与 Mn(2+)、Zn(2+)和 Cd(2+)的外排有关。afe_2126 编码的蛋白质可能与 Mn(2+)和 Zn(2+)外排以及 Cd(2+)摄取有关。