A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli |
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Authors: | Hwang S W Lee J H Park H B Pyo S H So J E Lee H S Hong S S Kim J H |
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Affiliation: | (1) Department of Chemical Engineering, Kongju National University, 182 Shinkwan-Donk, Kongju 314-701, Chungnam, South Korea |
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Abstract: | 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. |
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Keywords: | Antimicrobial peptide magainin derivative fusion protein purification hydroxylamine treatment chromatography |
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