Metchnikowin is a proline-rich peptide from Drosophila with antibacterial and antifungal activities. Its commercial application is limited due to the low immune-inducible expression in vivo. Here we present a method to produce high-yield metchnikowin in recombinant Escherichia coil. The genes coding for metchnikowin and the fusion partner Cherry tag were subcloned into the pET22b vector to construct a recombinant expression plasmid and transformed into E. coil BL21 (DE3). The fusion protein was successfully expressed and secreted with a yield of 300 mu g/ml after 18-h induction. Active metchnikowin was released by cleavage of the Asp-Pro bond between fused proteins by 72-h formic acid hydrolysis at 50 degrees C. After 24-h dialysis, metchnikowin was purified to electrophoretic homogeneity and showed significant antibacterial activities against both Bacillus subtilis and E. coil DH5 alpha. It is the first report on efficient production of metchnikowin in recombinant E. coli. (C) 2013 Elsevier Inc. All rights reserved.