| 根癌土壤杆菌SCUEC1菌株尼古丁降解代谢中agnE基因的克隆与功能鉴定 |
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| 引用本文： | 皮婷,姚家成,黄粤,夏珍珍,梅枫,何冬兰,程国军,刘涛,李晓华.根癌土壤杆菌SCUEC1菌株尼古丁降解代谢中agnE基因的克隆与功能鉴定[J].微生物学报,2018,58(5):907-914. |
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| 作者姓名： | 皮婷 姚家成 黄粤 夏珍珍 梅枫 何冬兰 程国军 刘涛 李晓华 |
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| 作者单位： | 中南民族大学生命科学学院微生物资源与利用湖北省工程技术研究中心 |
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| 基金项目： | 国家自然科学基金（31070087，30570046）；湖北省自然科学基金（2011CDA079，2008CDB076）；中央科研基本业务费（CZW16005，YCZW15104） |
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| 摘 要： | 【目的】本研究对尼古丁降解菌根癌土壤杆菌SCUEC1菌株中agnE基因进行克隆表达,并对agnE基因表达蛋白进行功能鉴定。【方法】通过PCR扩增获得agn 全长基因(1029 bp),构建重组质粒pET28a-agnE,转化大肠杆菌BL21(DE3)菌株进行异源表达,采用SDS-PAGE检测重组蛋白。以2,5-二羟基吡啶作为反应底物,在检测波长320 nm下测定反应液吸光度值,进一步研究温度、pH值和金属离子对AgnE蛋白酶活性的影响。【结果】克隆得到基因agnE,构建了p ET28a-agnE重组质粒并进行了表达,AgnE蛋白分子量为42.0 kDa,表达蛋白以包涵体形式存在于细胞中。酶促反应0、5、10、20 min时反应液吸光度分别为0.6170、0.2273、0.0907、0.0667。在pH 8.0、温度20°C下,AgnE蛋白具有较高的2,5-二羟基吡啶双加氧酶活性,且Fe2+对酶活性具有明显的促进作用。【结论】明确了Agn E蛋白具有2,5-二羟基吡啶双加氧酶活性。
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| 关 键 词： | 根癌土壤杆菌SCUEC1菌株 尼古丁降解代谢 agnE基因功能 2 5-二羟基吡啶 |
| 收稿时间： | 2017/10/22 0:00:00 |
| 修稿时间： | 2017/12/26 0:00:00 |
Cloning and charaterization of the agnE gene for nicotine degradation in Agrobacterium tumefaciens strain SCUEC1 |
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| Ting Pi,Jiacheng Yao,Yue Huang,Zhenzhen Xi,Feng Mei,Donglan He,Guojun Cheng,Tao Liu and Xiaohua Li.Cloning and charaterization of the agnE gene for nicotine degradation in Agrobacterium tumefaciens strain SCUEC1[J].Acta Microbiologica Sinica,2018,58(5):907-914. |
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| Authors: | Ting Pi Jiacheng Yao Yue Huang Zhenzhen Xi Feng Mei Donglan He Guojun Cheng Tao Liu and Xiaohua Li |
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| Institution: | Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China,Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China and Hubei Provincial Engineering and Technology Research Center for Resources and Utilization of Microbiology, College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, Hubei Province, China |
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| Abstract: | |
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| Keywords: | Agrobacterium tumefaciens SCUEC1 strain nicotine degrading metabolism the function of the agnE gene 2 5-dihydroxy pyridine |
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