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假单胞菌AD39菌株三嗪水解酶基因trzN的克隆、表达和酶鉴定
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作者 赵雅芳 赵化冰 +1 位作者 孙素菊 蔡宝立 《南开大学学报(自然科学版)》 CAS CSCD 北大核心 2012年第6期1-6,共6页
来自Pseudomonassp.AD39菌株的三嗪水解酶TrzN,能催化除草剂阿特拉津的水解脱氯反应,将阿特拉津转化成羟基阿特拉津.AD39菌株的TrzN以组氨酸标签蛋白的形式在大肠杆菌中表达,重组蛋白用镍柱亲和层析法纯化.纯化的TrzN蛋白能以阿特拉津... 来自Pseudomonassp.AD39菌株的三嗪水解酶TrzN,能催化除草剂阿特拉津的水解脱氯反应,将阿特拉津转化成羟基阿特拉津.AD39菌株的TrzN以组氨酸标签蛋白的形式在大肠杆菌中表达,重组蛋白用镍柱亲和层析法纯化.纯化的TrzN蛋白能以阿特拉津、阿特拉通、杀草净、扑草净、莠灭净和西玛津等6种三嗪类除草剂为催化底物,其中对莠灭净的催化活力最高.酶学实验表明,催化反应的最适pH为8.0,Co2+、Fe2+、Ca2+、Mg2+和Mn2+等金属离子对TrzN有激活作用,Zn2+、Cu2+和Ni2+等金属离子对TrzN的酶活力有抑制作用.酶动力学实验显示,TrzN蛋白的Km为41.47μmol.L-1,Vmax为2.48μmol.min-1mg-1. 展开更多
关键词 假单胞菌AD39 trzn表达 三嗪水解酶 酶学性质 酶动力学
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阿特拉津降解菌Arthrobacter sp. AG1降解基因研究 被引量:6
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作者 代先祝 蒋建东 +2 位作者 顾立峰 潘荣清 李顺鹏 《生物工程学报》 CAS CSCD 北大核心 2007年第5期789-793,共5页
菌株Arthrobacter sp. AG1能以4000mg/L的阿特拉津(AT)为唯一碳源、氮源和能源生长。通过设计特异引物从AG1中扩增出阿特拉津氯水解酶基因trzN的全序列,该基因与已报道的trzN基因序列相似性为99%。AG1菌株中含有两个大于100kb的质粒,Sou... 菌株Arthrobacter sp. AG1能以4000mg/L的阿特拉津(AT)为唯一碳源、氮源和能源生长。通过设计特异引物从AG1中扩增出阿特拉津氯水解酶基因trzN的全序列,该基因与已报道的trzN基因序列相似性为99%。AG1菌株中含有两个大于100kb的质粒,Southern杂交结果显示trzN和atzB基因均位于其中较大的一个质粒pAG1上。将AG1菌株在LB液体培养基中转接三代后,发现34%的细菌细胞丢失了降解活性,但却未发现丢失质粒,PCR扩增结果表明突变子丢失了trzN基因,但atzB和atzC基因未丢失,说明降解活性的缺失是trzN基因片段从质粒上丢失的结果,表明trzN基因在环境中存在水平转移现象,暗示菌株AG1中的阿特拉津降解基因是基因的水平转移重组的结果。 展开更多
关键词 ARTHROBACTER sp. AG1 trzn 基因水平转移 阿特拉津 降解
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Isolation and characterization of atrazine-degrading Arthrobacter sp. AD26 and use of this strain in bioremediation of contaminated soil 被引量:18
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作者 Li Qingyan Li Ying +1 位作者 Zhu Xikun Cai Baoli 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2008年第10期1226-1230,共5页
A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an ... A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewater sample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as an Arthrobacter sp. PCR assays indicated that AD26 contained atrazine-degrading genes trzN and atzBC. The trzN gene of AD26 only differs from the trzN of Arthrobacter aurescens TC1 by one base (A→T at 907) and one amino acid (Met→Leu at 303). The specific activity of trzN of AD26 in crude cell extract was 0.28 U/mg, which was 1.2 times that of TC1. This strain has shown faster growth and atrazine-degradation rates in atrazine-containing minimal media than two well characterized atrazine-degrading bacteria, Pseudomonas sp. ADP and Arthrobacter aurescens TC1. After incubating for 48 h at 30°C, the OD600 of AD26 reached 2.6 compared with 1.33 of ADP. AD26 was capable of degrading 500 mg/L of atrazine in minimal medium at 95% in 72 h, while the degradative rates by TC1 and ADP were only 90% and 86%, respectively. A bioremediation trial of contaminated soil has indicated that AD26 can degrade as high as 98% of atrazine contained in soil (300 mg/kg) after incubating for 20 d at 26°C, nominating this strain as a good candidate for use in bioremediation programs. 展开更多
关键词 ATRAZINE BIODEGRADATION Arthrobacter sp. AD26 trzn gene BIOREMEDIATION
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