Hwang S W, Lee J H, Park H B, Pyo S H, So J E, Lee H S, Hong S S, Kim J H
Department of Chemical Engineering, Kongju National University, 182 Shinkwan-Donk, Kongju 314-701, Chungnam, South Korea.
Mol Biotechnol. 2001 Jul;18(3):193-8. doi: 10.1385/MB:18:3:193.
A magainin derivative, designated MSI-344, was produced in Escherichia coli as fusion protein, by utilizing a truncated amidophsphoribosyltransferase of E. coli as a fusion partner. Bacterial cells transformed with the gene encoding the fusion protein were grown to a high cell density and induced with isopropyl-1-thio-b-D-galatoside (IPTG) to initiate product expression. The fusion protein was accumulated into cytoplasmic inclusion body and recombinant MSI-344 was released from the fusion partner by hydroxylamine treatment. Following cleavage of the fusion protein with hydroxylamine, the released MSI-344 was purified to homogeneity by cationic exchange chromatography. The final purity was at least 95% by reversed-phase high performance liquid chromatography (RP-HPLC). Purified recombinant MSI-344 was found to be indistinguishable from the synthetic peptide determined by amino acid sequences and antimicrobial activity assay.
