Heterologous expression of gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39

Yasushi Kawai, Kensuke Arakawa, Ayako Itoh, Boku Saitoh, Yasuyuki Ishii, Junko Nishimura, Haruki Kitazawa, Takatoshi Itoh, Tadao Saito

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14 Citations (Scopus)

Abstract

Gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39, has a cyclic structure linking N- and C-terminal amino acids. Gassericin A was expressed in Escherichia coli JM109 as a biotinylated fusion protein on the basis of the DNA sequence of mature bacteriocin. A positive clone accumulated the bacteriocin, with no activity, as a soluble fusion protein in the cytoplasm. After release of an N-terminal tag with factor Xa protease, gassericin A was converted into an active peptide having N- and C-termini. The total amount of purified bacteriocins (expressed and native) was 480 μg/L and 370 μg/L, respectively. However, the specific activity of expressed gassericin A was 15 AU/mg lower than that of native bacteriocin (2600 AU/mg). Although the actual Mr (molecular weight) of the expressed bacteriocin should be 5666, the peptide showed the same mobility (Mr 3800) in sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as native cyclic gassericin A, suggesting that the expressed peptide retains compact folding of the molecule similar to that of native gassericin A.

Original languageEnglish
Pages (from-to)45-51
Number of pages7
JournalAnimal Science Journal
Volume74
Issue number1
DOIs
Publication statusPublished - Feb 2003
Externally publishedYes

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Keywords

  • Bacteriocin
  • Cyclic
  • Expression
  • Gassericin A
  • Lactobacillus gasseri

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • veterinary(all)

Cite this

Kawai, Y., Arakawa, K., Itoh, A., Saitoh, B., Ishii, Y., Nishimura, J., Kitazawa, H., Itoh, T., & Saito, T. (2003). Heterologous expression of gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39. Animal Science Journal, 74(1), 45-51. https://doi.org/10.1046/j.1344-3941.2003.00085.x