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.
- Fuchs, P.C., Barry, A.L., and Brown, S.D. (1998) In vitro antimicrobial activity of MSI-78, a magainin analog. Antimicrob. Agents Chemother. 42, 1213–1216.
- Hancock, R. E. W. (1997) Peptide antibiotics. Lancet 349, 418–422. CrossRef
- Zasloff, M. (1987) Magainins, a class of antimicrobial peptides from Xenopus skin: isolation, characterization of two active forms, and partial cDNA sequence of a precursor. Proc. Natl. Acad. Sci. USA 84, 5449–5453. CrossRef
- Bowman, H. G., and Hultmark, D. (1987) Cell-free immunity in insects. Annu. Rev. Microbiol. 41, 103–126. CrossRef
- Ganz, T., Selsted, M. E., and Lehrer, R. I. (1990) Defensins. Eur. J. Haematol. 44, 1–8. CrossRef
- Habermann, E. (1972) Bee and wasp venoms. Science 177, 314–322. CrossRef
- Takahashi, H., Komano, H., Kawagushi, N., Obinata, M., and Nawri, S. (1984) Activation of secretion of specific proteins from the fat body following injury to the body wall of Sarcophaga peregrina larvae. Insect. Biochem. 14, 713–717. CrossRef
- Yarus, S., Rosen, J. M., Cole, A. M., Diamond, G. (1996) Production of active bovine tracheal antimicrobial peptide in milk of transgenic mice. Proc. Natl. Acad. Sci. USA 93, 14,118–14,121. CrossRef
- Anderson, D., Engsrom, A., Josephson, S., Hansson, L., and Steiner, H. (1991) Biologically active and amidated cecropin produced in a baculovirus expression system from a fusion construct containing the antibody-binding part of protein A. Biochem. J. 280, 219–224.
- Hellers, M., Gunne, H., Steiner, H. (1991) Expression and post-translational processing of preprocecropin A using a baculovirus vector. Eur. J. Biochem. 199, 435–439. CrossRef
- Reichart, J. M., Petit, I., Legrain, M., et al. (1992) Expression and secretion in yeast of active insect defensin, an inducible antibacterial peptide from the fleshlfy Phormia terranovae. Invertebr. Rerod. Dev. 21, 15–24.
- Hara, S., Yamakawa, M. (1996) Production in Escherichia coli of moricin, a novel type antibacterial peptide from the silkworm Bobvx mori. Biochem. Biophys. Res. Commun. 220, 664–669. CrossRef
- Haught, C., Davis, G. D., Subramanian, R., Jackson, K. W., Harrison, R. G. (1998) Recombinant production and purification of novel antisense antimicrobial peptide in Escherichia coli. Biotechnol. Bioeng. 57, 55–61. CrossRef
- Lee, J. H., Minn, I., Park, C. B., and Kim, S. C. (1998) Acidic peptide-mediated expression of the antimicrobial peptide buforin II as tandem repeats in Escherichia coli. Protein Expr. Purif. 12, 53–60. CrossRef
- Zhang, L., Falla, T., Wu, M., et al. (1998) Determinants of recombinant production of antimicrobial cationic peptides and creation of peptide variants in bacteria. Biochem. Biophys. Res. Commun. 247, 674–680. CrossRef
- Datar, R. V., Cartwright, T., and Rosen, C. G. (1993) Process economics of animal cell and bacterial fermentations: A case study analysis of tissue plasminogen activator. Biotechnology 11, 349–357. CrossRef
- Maloy, W. L., and Kari, U. P. (1997) Structure-activity studies on magainins and other host defense peptides. Biopolymers 37, 105–122. CrossRef
- Friedrich, C., Scott, M. G., Karunaratne, N., Yan, H., and Hancock, R. E. W. (1999) Salt-resistant alpha-helical cationic antimicrobial peptides. Antimicrob. Agents. Chemother. 43, 1542–1548.
- Tso, J. Y., Hermodson, M. A., Zalkin, H. (1982) Glutamine phosphoribosylpyro- phosphate amidotransferase from cloned Escherichia coli purF. NH2-terminal amino acid sequence, identification of the glutamine site, and trace metal analysis. J. Biol. Chem. 257, 3532–3536.
- Borstein, P. and Balian, G. (1977) Cleavage at Asn-Gly bonds with hydroxylamine. Methods Enzymol. 67, 132–145. CrossRef
- A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli
Volume 18, Issue 3 , pp 193-198
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- Humana Press
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- Antimicrobial peptide
- magainin derivative
- fusion protein
- hydroxylamine treatment
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