Cloning and heterologous expression of a bacteriocin sakacin P from Lactobacillus sakei in Escherichia coli

Appl Microbiol Biotechnol. 2012 May;94(4):1061-8. doi: 10.1007/s00253-012-3872-z.

Abstract

Sakacin P, a bacteriocin from Lactobacillus sakei, shows strong activity against food-borne pathogens such as Listeria monocytogenes. In L. sakei, the structural gene (sppA) encoding sakacin P is controlled by a strict regulatory mechanism, and the quantity of secreted sakacin P is limited. In this study, the sppA gene was synthesized by splicing overlap extension PCR and cloned into Escherichia coli. After the induction with isopropyl-β-d-thiogalactopyranoside, the recombinant sakacin P was successfully expressed. The collected cells were sonicated, and the activity was detected by agar diffusion method. The results also showed that the low-temperature induction can improve the activity of sakacin P.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacteriocins / genetics*
  • Bacteriocins / pharmacology*
  • Cloning, Molecular*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Listeria monocytogenes / drug effects
  • Microbial Sensitivity Tests

Substances

  • Anti-Bacterial Agents
  • Bacteriocins
  • sakacin P protein, Lactobacillus