| 来源于Pseudomonas sp.1 7对硝基酚降解基因簇中的对苯二醌还原酶基因的克隆和功能鉴定 |
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| 引用本文： | 张双宇,孙雯,许丽,田健,初晓宇,伍宁丰.来源于Pseudomonas sp.1 7对硝基酚降解基因簇中的对苯二醌还原酶基因的克隆和功能鉴定[J].中国农业科技导报,2012,14(6):70-75. |
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| 作者姓名： | 张双宇 孙雯 许丽 田健 初晓宇 伍宁丰 |
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| 作者单位： | (中国农业科学院生物技术研究所, 北京 100081) |
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| 基金项目： | 国家自然科学基金项目(31170036)资助 |
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| 摘 要： | Pseudomonas sp. 1 7 可以利用对硝基酚（PNP）作为唯一的碳源，氮源和能源生长。为了鉴定菌株1 7代谢PNP的相关基因，本研究构建了菌株1 7基因组的Fosmid文库，并通过文库的筛选克隆得到一段长为12.3 kb的基因簇序列。序列分析显示，该基因簇中共有7个基因（pdcEDGFCBA）与PNP的代谢相关。其中PdcB与来源于Pseudomonas sp. WBC 3中的对苯二醌还原酶（PnpB）有98%的相似性。将pdcB基因在E. coli BL21中过量表达，并通过Ni NTA亲和层析纯化重组蛋白PdcB。体外的酶活测定表明，PdcB为一个FAD和NADH依赖型的对苯二醌还原酶，能够催化对苯二醌降解并生成对苯二酚。
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| 关 键 词： | 假单胞菌 对硝基酚 生物修复 对苯二醌还原酶 |
Gene Cloning and Functional Identification of p-benzoquinone Reductase from Pseudomonas sp.1-7: A Novel Member of p-Nitrophenol Degradation Gene Cluster |
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| ZHANG Shuang-yu,SUN Wen,XU Li,TIAN Jian,CHU Xiao-yu,WU Ning-feng.Gene Cloning and Functional Identification of p-benzoquinone Reductase from Pseudomonas sp.1-7: A Novel Member of p-Nitrophenol Degradation Gene Cluster[J].Journal of Agricultural Science and Technology,2012,14(6):70-75. |
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| Authors: | ZHANG Shuang-yu SUN Wen XU Li TIAN Jian CHU Xiao-yu WU Ning-feng |
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| Institution: | (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
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| Abstract: | Pseudomonas sp. 1 7 utilizes p nitrophenol (PNP) as a sole source of carbon, nitrogen, and energy. In order to identify the genes involved in this utilization, a gene cluster was identified in a 12.3 kb DNA fragment of a genome fosmid library. Sequence analysis revealed that there were 7 genes (pdcEDGFCBA) related in PNP degradation. PdcB shares 98% identity to the p benzoquinone reductase (PnpB) from Pseudomonas sp. WBC 3. The pdcB gene was then over expressed in E.coli BL21, and the recombinant protein PdcB was further purified with Ni NTA affinity chromatography. Purified PdcB was shown a FAD and NADPH dependent p benzoquinone reductase that catalyzed the reduction of p benzoquinone to hydroquinone by in vitro activity assays. |
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| Keywords: | Pseudomonas p nitrophenol bioremediation p benzoquinone reductase |
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