Issue 19, 2020

A simplified protein purification method through nickel cleavage of the recombinant protein from the Escherichia coli cell surface

Abstract

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.

Graphical abstract: A simplified protein purification method through nickel cleavage of the recombinant protein from the Escherichia coli cell surface

Supplementary files

Article information

Article type
Communication
Submitted
27 मई 2020
Accepted
21 जुलाई 2020
First published
22 जुलाई 2020

Analyst, 2020,145, 6227-6231

A simplified protein purification method through nickel cleavage of the recombinant protein from the Escherichia coli cell surface

S. Huang, T. Wei, W. Sha, Q. Hu, Y. Zhang, J. Wang, Y. Jiang and H. Chen, Analyst, 2020, 145, 6227 DOI: 10.1039/D0AN01060J

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