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Heterologous expression of gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39
Author(s) -
KAWAI Yasushi,
ARAKAWA Kensuke,
ITOH Ayako,
SAITOH Boku,
ISHII Yasuyuki,
NISHIMURA Junko,
KITAZAWA Haruki,
ITOH Takatoshi,
SAITO Tadao
Publication year - 2003
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1046/j.1344-3941.2003.00085.x
Subject(s) - bacteriocin , lactobacillus gasseri , escherichia coli , peptide , biochemistry , protease , chemistry , gel electrophoresis , fusion protein , lactobacillus , biology , microbiology and biotechnology , antimicrobial , enzyme , fermentation , recombinant dna , gene
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.