Active, Soluble Recombinant Melittin Purified by Extracting Insoluble Lysate of Escherichia coli Without Denaturation

作者：Buhrman Jason S; Rayahin Jamie E; Cook Laura C; Federle Michael J; Gemeinhart Richard A^*

来源：Biotechnology Progress, 2013, 29(5): 1150-1157.

DOI：10.1002/btpr.1784

• recombinant protein
• fusion protein
• protein extraction
• surfactant
• melittin

Cell lytic peptides are a class of drugs that can be used to selectively kill invading organisms or diseased cells. Several of these peptides have been identified as potential therapeutics. Herein, we report a novel process for purifying recombinant melittin, a cell lytic peptide that inserts into the membranes of cells causing cell lysis, from Escherichia coli. The process involves surfactant and low pH to solubilize melittin fusion proteins from the insoluble fraction of bacterial lysates. We are able to significantly improve purity of the final product and confirm the activity of the peptide. The process yields recombinant melittin that is effective when used to treat U-87 MG glioma cells and inhibits growth of the gram-positive pathogenic bacterium Streptococcus pyogenes. We demonstrate a method of repeated extraction of the insoluble protein fraction with mild detergent at a low pH that is able to generate a yield of pure, soluble melittin of approximate to 0.5-1 mg/L of E. coli culture.

• 出版日期2013-9