Lanthionine introduction into nukacin ISK-1 prepeptide by co-expression with modification enzyme NukM in Escherichia coli
| Autor: | Yoshitaka Harada, Takeshi Zendo, Kenji Sonomoto, Kouki Shioya, Jiro Nakayama, Yuji Aso, Jun Ichi Nagao |
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| Rok vydání: | 2005 |
| Předmět: |
Signal peptide
Molecular Sequence Data Biophysics Peptide Sulfides Protein Engineering Biochemistry chemistry.chemical_compound Bacteriocins Escherichia coli Amino Acid Sequence Protein precursor Molecular Biology Peptide sequence Hydro-Lyases Lanthionine Alanine chemistry.chemical_classification Escherichia coli Proteins Gene Expression Regulation Bacterial Cell Biology Lantibiotics Recombinant Proteins Amino acid Molecular Weight chemistry Peptides |
| Zdroj: | Biochemical and Biophysical Research Communications. 336:507-513 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2005.08.125 |
| Popis: | 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 72Da 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. |
| Databáze: | OpenAIRE |
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