HPLC purification and re-evaluation of chemical identity of two circular bacteriocins, gassericin A and reutericin 6

Kensuke Arakawa, Y. Kawai, Y. Ito, K. Nakamura, T. Chujo, J. Nishimura, H. Kitazawa, T. Saito

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Aim: The study aimed for the complete purification and recharacterization of the highly hydrophobic circular bacteriocins, gassericin A and reutericin 6. Methods and Results: Gassericin A and reutericin 6 were purified to homogeneity using previously described method and reverse-phase HPLC with an octyl column and eluents of aqueous acetonitrile and 2-propanol. Mass analysis, N-terminal sequencing and bacteriocin assay of the HPLC-purified bacteriocins showed the two bacteriocins had identical seamless circular structures with the same m/z value (5651) of [M + H]+ and both had the same specific activity. d/l-amino acid composition analysis using two distinct methods with the chiral fluorescent derivatization reagents (+)-1-(9-fluorenyl)ethyl chloroformate and o-phthalaldehyde/N-acetyl-l-cystein revealed neither gassericin A nor reutericin 6 contained d-alanine residues contrary to our previous results. Conclusion: Purified gassericin A and reutericin 6 are chemically identical circular molecules containing no d-alanine residues. Significance and Impact of the Study: The HPLC conditions developed in this study will facilitate advanced purification and correct characterization of other highly hydrophobic bacteriocins.

Original languageEnglish
Pages (from-to)406-411
Number of pages6
JournalLetters in Applied Microbiology
Volume50
Issue number4
DOIs
Publication statusPublished - Apr 2010
Externally publishedYes

Fingerprint

Bacteriocins
High Pressure Liquid Chromatography
Alanine
o-Phthalaldehyde
2-Propanol
gassericin A
Amino Acids

Keywords

  • Bacteriocin
  • Circular peptide
  • Gassericin A
  • HPLC purification
  • Lactobacillus gasseri
  • Lactobacillus reuteri
  • Reutericin 6

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

Cite this

HPLC purification and re-evaluation of chemical identity of two circular bacteriocins, gassericin A and reutericin 6. / Arakawa, Kensuke; Kawai, Y.; Ito, Y.; Nakamura, K.; Chujo, T.; Nishimura, J.; Kitazawa, H.; Saito, T.

In: Letters in Applied Microbiology, Vol. 50, No. 4, 04.2010, p. 406-411.

Research output: Contribution to journalArticle

Arakawa, Kensuke ; Kawai, Y. ; Ito, Y. ; Nakamura, K. ; Chujo, T. ; Nishimura, J. ; Kitazawa, H. ; Saito, T. / HPLC purification and re-evaluation of chemical identity of two circular bacteriocins, gassericin A and reutericin 6. In: Letters in Applied Microbiology. 2010 ; Vol. 50, No. 4. pp. 406-411.
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