| Autor: |
Huang, Shanqing, Wei, Tianbiao, Sha, Wanxing, Hu, Qingyuan, Zhang, Yingying, Wang, Jue, Jiang, Yufei, Chen, Hao |
| Předmět: |
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| Zdroj: |
Analyst; 10/7/2020, Vol. 145 Issue 19, p6227-6231, 5p |
| Abstrakt: |
To simplify the protein purification process, we developed a novel one-step purification method in which the recombinant protein can be cleaved directly from the Escherichia coli cell surface. This method involves fusion of the target protein to the C-terminus of a LOS tag comprising a surface anchor protein (Lpp-OmpA) and a sequence-specific nickel-assisted cleavage (SNAC)-tag. The LOS tag facilitates the anchoring of the target protein to the outer membrane of E. coli cells and its separation from the cell membrane through Ni2+ cleavage. Intact, biologically active protein with a purity of 95% and a yield of approximately 100 mg per liter of culture can be readily obtained through Ni2+ cleavage in resuspension solution followed by centrifugation. In this study, a practical and promising protein purification method has been established with minimal labor and cost, as no cell disruption and chromatographic separation are required downstream. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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