A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli
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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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- A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli
Volume 18, Issue 3 , pp 193-198
- Cover Date
- Print ISSN
- Online ISSN
- Humana Press
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- Antimicrobial peptide
- magainin derivative
- fusion protein
- hydroxylamine treatment
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