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. 2022:2406:339-358.
doi: 10.1007/978-1-0716-1859-2_20.

Optimizing Chaperone Removal Strategy from Overexpressed Recombinant Proteins : GNE, a Case Study

Shweta Sharma  1 , Roop Singh Bora  2 , Kulvinder Singh Saini  3 , Ranjana Arya  4
Affiliations

Optimizing Chaperone Removal Strategy from Overexpressed Recombinant Proteins : GNE, a Case Study

Shweta Sharma et al. Methods Mol Biol. 2022.

Abstract

In the last two decades, numerous innovative advances, strategies and protocols have been developed and optimized to improve the quality and quantity of soluble recombinant protein production in E. coli. One of the major challenges being the coelution of chaperone proteins along with desired recombinant protein of interest. The removal of chaperones is important for protein yield, structural determination, optimal activity, and desired function of the recombinant protein. In this chapter, we outline various strategies for removal of chaperone contaminants from oligomeric proteins, with the ultimate objective of ameliorating the quality and proper folding of recombinant proteins. We have discussed in detail the purification and expression of full-length protein, GNE (UDP-N-acetylglucosamine 2-epimerase/ N-acetylmannosamine kinase), as a case study for chaperone removal.

Keywords: Chaperones; DnaK; GST Cleanser Protein; Protein solubility.

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