文献类型：期刊文献

英文题名：Novel Application of Magnetic Protein: Convenient One-Step Purification and Immobilization of Proteins

作者：Jiang, Min[1];Zhang, Lujia[2,3];Wang, Fengqing[1];Zhang, Jie[4];Liu, Guosong[1];Gao, Bei[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China;[2]East China Normal Univ, Sch Chem & Mol Engn, Shanghai 200062, Peoples R China;[3]NYU Shanghai, NYU ECNU Ctr Computat Chem, Shanghai 200062, Peoples R China;[4]East China Normal Univ, Sch Phys & Mat Sci, Shanghai 200062, Peoples R China

年份：2017

卷号：7

外文期刊名：SCIENTIFIC REPORTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000413084800026)】；

基金：This work was funded by Natural Science Foundation of Shanghai (No. 16ZR1449500), the National High Technology Research and Development Program of China (No. 2013AA102109), and the National Natural Foundation of China (No. 31571786).

语种：英文

摘要：Recently, a magnetic protein was discovered, and a multimeric magnetosensing complex was validated, which may form the basis of magnetoreception. In this study, the magnetic protein was firstly used in biotechnology application, and a novel convenient one-step purification and immobilization method was established. A universal vector and three linker patterns were developed for fusion expression of magnetic protein and target protein. The magnetic protein was absorbed by iron beads, followed by target protein aggregation, purification, and immobilization. GFP, employed as a reporter protein, was successfully purified from cell lysate. Subsequently, three enzymes (lipase, a-L-arabinofuranosidase, pullulanase) with different molecular sizes testified the versatility of this magnetic-based approach. The specific activities of the purified enzymes were distinctly higher than those of the traditionally purified enzymes using affinity chromatography. The lipase immobilized on iron beads presented improved thermostability and enhanced pH tolerance compared to the free enzyme. The immobilized lipase could be easily recovered and reused for maximum utilization. After 20 cycles of reutilization, the magnetically immobilized lipase retained 71% of its initial activity. This investigation may help introduce magnetic protein into biotechnology applications, and the one-step purification and immobilization method may serve to illustrate an economically viable process for industry.