Biopolymers and cell. Volume 20. № 3. 182-192.

P. V. Gilchuk

Evaluation of renaturation methods for industrial obtaining of recombinant proteins from Escherichia coli inclusion bodies in biologically active form

Summary

One of the widely used strategies for the production of recombinant proteins in Escherichia coli cells is their accumulation as insoluble cytoplasmatic aggregates-inclusion bodies-during the expression. The advantage of such synthesis is determined by the high leves of accumulation and purity of the target protein as well as by the possibility to carry out successful renaturation and to obtain active product from insoluble aggregates under conditions in vitro. The main problem hindering industrialization of this process is caused by the absence of universal renaturation schemes, as refolding of the recombinant proteins is an empiric process and needs the development of unique protocols for individual proteins. Study on main aggregation modes and folding processes in vivo allowed developing effective approaches for the refolding of many recombinant proteins. The evaluation of some contemporary methods of protein refolding is given in the review. The main strategies, that can be used for the industrial obtaining of proteins from bacterial inclusion bodies, are discussed. Recent achievements in the area of refolding recombinant proteins that contain disulfide bonds (oxidative refolding) are briefly described. The reasons of protein aggregation in the process of in vitro renaturation as well as the approaches used for increasing yield of correctly refolded protein are analyzed.