焦化废水活性污泥PAH双加氧酶基因多样性分析.pdf

中国环境科学 2017,37(1) ：367~372 China Environmental Science 焦化废水活性污泥 PAH 双加氧酶基因多样性分析 蒙小俊 1,2,李海波 2,盛宇星 2,曹宏斌 2* (1.安康学院旅游与资源环境学院,陕西 安康 725000 ；2. 中国科学院过 程工程研究所绿色过程与工程重点实验 ,北京 100190) 摘要：为分析焦化废水活性污泥中降解 PAH 双加氧酶的多样性,利用 16Sr DNA-PCR-DGGE 方法, 以实际焦化废水好氧单元活性污泥总 DNA 为模板,通过引物对污泥中的双加氧酶基因进行了克隆表达和多样性分析.结果表明, 以活性污泥总 DNA 为模板,利用引物 RHD-GN-610F 和 RHD-GN-916R 扩增后有明显的产物,产物大小约为 300bp;DGGE 指纹图谱显示 PCR 产物有 8 条分离条带,丰度分别为 2.99% 、8.16%、20.75% 、28.50% 、8.62%、7.26%、10.62%和 13.10%;条带经切胶回收 PCR 扩增后出现明显的扩增产物,TA 克隆后成功测 试出 4 条序列,长度分别为 305bp,298bp,334bp 和 294bp,表明焦化废水活性污泥中存在不同降解 PAH 的 RHD 酶.这些结果为焦化废水中 PAHs 的风险评估及其潜在生物降解提供理论基础. 关键词：焦化废水；PAHs ；活性污泥；双加氧酶基因 中图分类号：X703 文献标识码：A 文章编号：1000-6923(2017)01-0367-06 1,2 2 Diversity of PAH dioxygenase genes of activated sludge from coking wastewater. MENG Xiao-jun , LI Hai-bo , SHENG Yu-xing2, CAO Hong-bin2*(1.School of Tourism and Environment, Ankang University, Ankang 725000, China ； 2.Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China). China Environmental Science, 2017,37(1)：367~372 Abstract ：In order to analyze the biodiversity of PAH dioxygenase in activated sludge from a full-scale coking wastewater, primers were used for dioxygenase gene cloning expression and diversity analysis by 16Sr DNA-PCR-DGGE method using total DNA of aerobic activated sludge as the template. The results showed that significant amplification product was found using primers RHD-GN-610F and RHD-GN-916R, and the product size was 300bp. PCR product was conducted by DGGE analysis and eight separate bands were found, the abunda