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KMID : 1094720150200050901
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Biotechnology and Bioprocess Engineering
2015 Volume.20 No. 5 p.901 ~ p.907
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One-step purification and immobilization of his-tagged protein via Ni2+-functionalized Fe3O4@polydopamine magnetic nanoparticles
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Yang Jianbing
Ni Kefeng
Wei Dongzhi
Ren Yuhong
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Abstract
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Ni2+-functionalized Fe3O4@polydopamine magnetic nanoparticles (Ni2+-PD-MNPs) were designed and synthesized by in situ coating of magnetic nanoparticles with polydopamine, followed by conjugation of Ni2+ to the polydopamine film. The Ni2+-PD-MNPs were used to purify His-tagged red fluorescent protein (His-RFP) via affinity interaction between Ni2+ and the His-tag. The results showed that the Ni2+-PD-MNPs had extraordinary selectivity for His-RFP purification. In addition, a Histagged transaminase (ω-transaminase BJ110) was selectively immobilized onto the Ni2+-PD-MNPs without purification, and the immobilized enzyme showed improved specific activity, as well as enhanced stability and reusability.
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KEYWORD
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His-tagged protein, immobilization, magnetic nanoparticles, polydopamine, purification
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