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

Biotechnol Appl Biochem. 2007 Aug;47(Pt 4):185-9. doi: 10.1042/BA20060214.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels / chemistry
  • Chromatography, Affinity / methods*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Humans
  • Metals / chemistry
  • Profilins / chemistry*
  • Profilins / isolation & purification*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification

Substances

  • Affinity Labels
  • Metals
  • Profilins
  • Recombinant Proteins