Lanthionine introduction into nukacin ISK-1 prepeptide by co-expression with modification enzyme NukM in Escherichia coli

Jun Ichi Nagao, Yoshitaka Harada, Kouki Shioya, Yuji Aso, Takeshi Zendo, Jiro Nakayama, Kenji Sonomoto

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

We demonstrated lanthionine introduction into hexa-histidine-tagged (His-tagged) nukacin ISK-1 prepeptide NukA by modification enzyme NukM in Escherichia coli. Co-expression of nukA and nukM, purification of the resulting His-tagged prepeptide by affinity chromatography, and subsequent mass spectrometry analysis showed that the prepeptide was converted into a postulated peptide with decrease in mass of 72 Da which resulted from dehydration of four amino acids. Characterization of the resultant prepeptide indicated the presence of unusual amino acids, such as dehydrated amino acid, lanthionine or 3-methyllanthionine, in its C-terminal propeptide moiety. The modified prepeptide encompassing the leader peptide attached to the post-translationally modified propeptide moiety was readily obtained by one-step purification. Our findings will thus be a powerful tool for introducing unusual amino acids aimed at peptide engineering and also helpful to provide new insight for further understanding of lanthionine-forming enzymes for lantibiotics.

Original languageEnglish
Pages (from-to)507-513
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume336
Issue number2
DOIs
Publication statusPublished - Oct 21 2005

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Escherichia coli
Amino Acids
Enzymes
Histidine
Purification
Affinity chromatography
Bacteriocins
Peptides
Protein Sorting Signals
Dehydration
Affinity Chromatography
Mass spectrometry
Mass Spectrometry
nukacin ISK-1
lanthionine

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Lanthionine introduction into nukacin ISK-1 prepeptide by co-expression with modification enzyme NukM in Escherichia coli. / Nagao, Jun Ichi; Harada, Yoshitaka; Shioya, Kouki; Aso, Yuji; Zendo, Takeshi; Nakayama, Jiro; Sonomoto, Kenji.

In: Biochemical and Biophysical Research Communications, Vol. 336, No. 2, 21.10.2005, p. 507-513.

Research output: Contribution to journalArticle

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AU - Aso, Yuji

AU - Zendo, Takeshi

AU - Nakayama, Jiro

AU - Sonomoto, Kenji

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