| TRAIL蛋白的体外活性稳定性初步研究 |
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| 引用本文： | 张红,魏东芝,周文瑜,陶欣艺,沈亚领,周劲松,张国钧,孙菁,球谊.TRAIL蛋白的体外活性稳定性初步研究[J].工业微生物,2011,41(1):9-14. |
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| 作者姓名： | 张红 魏东芝 周文瑜 陶欣艺 沈亚领 周劲松 张国钧 孙菁 球谊 |
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| 作者单位： | 1. 华东理工大学生物反应器工程国家重点实验室,上海,200237
2. 上海恰尔生物技术有限公司,上海,201103 |
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| 基金项目： | 上海市重点学科建设项目资助 |
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| 摘 要： | 大肠杆菌发酵表达的TRAIL蛋白经过两步纯化可得到纯度95%以上的活性蛋白,得率为40%.天然TRAIL蛋白为同源三聚体形式,在三聚体中心隐蔽结合Zn<'2+>,Zn<'2+>可维持TRAIL蛋白天然结构的稳定性.研究发现在纯化后的重组可溶性TRAIL蛋白中添加Zn<'2+>,并且Zn<'2+>与TRAIL单体的摩尔比...
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| 关 键 词： | TRAIL 纯化 Zn2+ L-Arg L-Glu 蛋白稳定性 |
Preliminary studies on long term stable activity of TRAIL protein in vitro |
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| ZHANG Hong,WEI Dong-Zhi,WEN Yu-Zhou,XIN Yi-Tao,SHEN Ya-Ling,ZHOU Jin-Song,Zhang Guo-jun,SUN Jing,QIU Yi.Preliminary studies on long term stable activity of TRAIL protein in vitro[J].Industrial Microbiology,2011,41(1):9-14. |
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| Authors: | ZHANG Hong WEI Dong-Zhi WEN Yu-Zhou XIN Yi-Tao SHEN Ya-Ling ZHOU Jin-Song Zhang Guo-jun SUN Jing QIU Yi |
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| Institution: | 1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; 2. Shanghai TRAIL bio-technical Co., LTD., Shanghai 201103, China ) |
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| Abstract: | A simple and efficient process of two-step purification of soluble TRAIL protein was established. The purity of 95% and the total recovery of 40% were obtained. This process was suitable for industrialized manufacturing crystallographic. Studies showed that the soluble TRAIL had a homotrimeric structure, and an internal zinc atom was bound by the cysteine residues at position 230 of each subunit, which was crucial for trimer stability and biologic activity. TRAIL was not a stable protein, but a phenomenon was demonstrated that the addition Zn^2＋ of molar ratio to TRAIL monomer with 2 ： 1 in purified TRAIL solution and simultaneous addition of 25 mmol/L L-Arg and 50 mmol/L L-GIu to the purified TRAIL could dramatically stabilize its activity and reduce aggregation and precipitation. |
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| Keywords: | TRAIL Zn2+ L-Arg L-Glu |
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