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Sequencing and expression analysis of sakacin genes in Lactobacillus curvatus strains

Abstract

In this study, we focused our investigation on two strains of Lactobacillus curvatus, L442 and LTH1174, which are able to produce bacteriocins. L. curvatus LTH1174 is widely studied for its capability to produce curvacin A, while L. curvatus L442 was isolated from traditional Greek fermented sausages and was shown to possess a strong inhibitory activity toward Listeria monocytogenes. By polymerase chain reaction, we were able to target in both strains the genes for the production of sakacin P and sakacin Q, sppA and sppQ, respectively, both encoded chromosomally. While sppA was found to be conserved when compared with other sakacin P genes, sppQ showed a deletion of about 15 nucleotides when aligned with sequences obtained from Lactobacillus sakei. This difference did not affect the activity of sakacin Q as determined by testing sensitive strains. Expression analysis highlighted that sakacin P was expressed in L. curvatus L442 but not in L. curvatus LTH1174. Curing experiments were performed on L. curvatus LTH1174 to study the effect of the megaplasmid, present in this strain. In the plasmid-cured strain, expression of the sppA gene was detected. sppQ was expressed in both plasmid-cured and wild-type L. curvatus LTH1174, although expression was higher in the plasmid-cured strain.

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Acknowledgments

This research has been funded by the E. C. within the framework of the specific research and technological development program “Confirming the International Role of the Community Research,” (contract no. ICA4-CT-2002-10037). We are grateful to Prof. Walter P. Hammes, University of Hohneheim, Stuttgart, Germany, and to Prof. John Metaxopoulos, Agricultural University of Athens, Greece, for the strains L. curvatus LTH1174 and 442, respectively.

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Correspondence to Luca Cocolin.

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Cocolin, L., Rantsiou, K. Sequencing and expression analysis of sakacin genes in Lactobacillus curvatus strains. Appl Microbiol Biotechnol 76, 1403–1411 (2007). https://doi.org/10.1007/s00253-007-1120-8

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Keywords

  • Bacteriocins
  • Expression analysis
  • Gene sequencing
  • Bioprotection