One-step purification of histidine-tagged profilin with high purity and yield by using metal precipitation.

A simple one-step method for the purification of recombinant His-tagged profilin from the bacterial cell lysate is reported. Noting the greater ease with which hexahistidine-tagged proteins can be metalprecipitated compared with unwanted protein impurities, we investigated the effect of lysis-buffer additives and optimization of other conditions to recover selectively desired proteins in a one-step metal precipitation without using biopolymers. Purification of the His-tagged melon (Cucumis melo) profilin was used to demonstrate the utility of this method and up to 80% recovery with a purity of 98% was achieved. This method obtained a yield of the protein nearly comparable with that obtained using metal-affinity chromatography. This purification procedure can reduce the time and cost of the purification process, especially on a large scale.