Enhancing the Purification and Stability of Superoxide Dismutase by Fusion with Thermoresponsive Self‐Assembly of Elastin Like Polypeptide

Abstract

Elastin‐like polypeptide (ELP) is a thermo‐sensitive biosynthetic polymer composed of a Val‐Pro‐Gly‐Xaa‐Gly repeating unit possessing a sharp reversible phase transition property at specific temperatures. Here, ELP was fused to human superoxide dismutase 1 (hSOD1) modified with His tag to produce recombinant hSOD1‐Linker‐ELP‐His (hSODLEH) which was expressed in E. coli and purified via inverse thermal cycling (ITC) and Ni‐NTA resin. The results showed that ELP tag did not affect the soluble expression of SOD1. The purification by ITC was superior to Ni‐NTA resin due to its convenient purification process, improved recovery rate and purification fold, thus indicating industrial suitability for large scale protein purification in a cost‐ and time‐efficient manner. Moreover, one round of ITC was sufficient for the recovery of highly purified recombinant SOD. The results further showed that ELP improved the stability of the SOD, and did not affect its secondary structure nor its ability to bind divalent metal ions. Overall, ELP‐protein fusion system could be a promising tool for large scale enzyme purification.