Effect of Amino Acid Substitutions on the Activity of Carnobacteriocin B2

  • Quadri L
  • Yan L
  • Stiles M
  • et al.
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Abstract

Carnobacteriocin B2, a 48-amino acid antimicrobial peptide containing a YGNGV motif that is produced by the lactic acid bacterium Carnobacterium piscicola LV17B, was overexpressed as fusion with maltose-bind- ing protein in Escherichia coli. This fusion protein was cleaved with Factor Xa to allow isolation of the mature bacteriocin that was identical in all respects to that obtained from C. piscicola. Similar methodology permit- ted production of the precursor precarnobacteriocin B2 (CbnB2P), which has an 18-amino acid leader, as well as six mutants of the mature peptide: CbnF3 (Tyr3 3 Phe), CbnS33 (Phe33 3 Ser), CbnI34 (Val34 3 Ile), CbnI37 (Val37 3 Ile), CbnG46 (Arg46 3 Gly), and Cbn28 (truncat- ed frameshift mutation: (carnobacteriocin B2 1–28) ⴙ ELTHL). Examination of these compounds for antimi- crobial activity showed that although CbnI34, CbnI37, and CbnG46 were fully active, CbnB2P, CbnF3, CbnS33, Cbn28, and all of the fusion proteins had greatly re- duced or no antimicrobial activity. Expression of the immunity protein that protects against the action of the parent carnobacteriocin B2 in a previously sensitive or- ganism also protects against the active mutants. Be- cause carnobacteriocin B2 also acts as an inducer of bacteriocin production in C. piscicola, the ability of the precursor CbnB2P and the mutants to exert this effect was examined. All were able to induce Bacⴚ cultures and reestablish the Bacⴙ phenotype except for the truncated Cbn28. The results demonstrate that very minor changes in the peptide sequence may drastically alter antimicrobial activity but that the induction of bacteri- ocin production is much more tolerant of structural modification, especially at the N terminus

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APA

Quadri, L. E. N., Yan, L. Z., Stiles, M. E., & Vederas, J. C. (1997). Effect of Amino Acid Substitutions on the Activity of Carnobacteriocin B2. Journal of Biological Chemistry, 272(6), 3384–3388. https://doi.org/10.1074/jbc.272.6.3384

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