Dataset of "Single-step Purification and Characterization of Pseudomonas aeruginosa Azurin"

Azurin is a small periplasmic blue copper protein. We have created a novel approach for expressing and purifying of azurin in E. coli with high yields and optimal metalation ratio. The azurin sequence was N-terminally fused with a GST tag and protein was purified by single-step affinity chromatography on a GST-trap column. The N-terminal tag was cleaved off by HRV 3C protease and sufficient metalation was endured by incubation with copper sulphate. UV-VIS absorption, mass spectroscopy, and circular dichroism analysis all validated the effective production of azurin, appropriate protein folding and the development of an active site with an associated cofactor. MD simulations verified that incorporation of the N-terminal GPLGS segment does not affect the wild-type azurin structure.