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

Research output: Contribution to journalArticle

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

Fingerprint

Lactobacillus gasseri
Bacteriocins
bacteriocins
peptides
Peptides
Native Polyacrylamide Gel Electrophoresis
Factor Xa
polyacrylamide gel electrophoresis
gassericin A
Cytoplasm
Proteins
Peptide Hydrolases
proteinases
cytoplasm
proteins
Clone Cells
Molecular Weight
Sodium
sodium
clones

Keywords

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

ASJC Scopus subject areas

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

Cite this

Heterologous expression of gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39. / Kawai, Yasushi; Arakawa, Kensuke; Itoh, Ayako; Saitoh, Boku; Ishii, Yasuyuki; Nishimura, Junko; Kitazawa, Haruki; Itoh, Takatoshi; Saito, Tadao.

In: Animal Science Journal, Vol. 74, No. 1, 02.2003, p. 45-51.

Research output: Contribution to journalArticle

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, vol. 74, no. 1, pp. 45-51. https://doi.org/10.1046/j.1344-3941.2003.00085.x
Kawai, Yasushi ; Arakawa, Kensuke ; Itoh, Ayako ; Saitoh, Boku ; Ishii, Yasuyuki ; Nishimura, Junko ; Kitazawa, Haruki ; Itoh, Takatoshi ; Saito, Tadao. / Heterologous expression of gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39. In: Animal Science Journal. 2003 ; Vol. 74, No. 1. pp. 45-51.
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