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Journal of Bacteriology, July 2004, p. 4276-4284, Vol. 186, No. 13
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.13.4276-4284.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Heterologous Expression and Purification of Active Divercin V41, a Class IIa Bacteriocin Encoded by a Synthetic Gene in Escherichia coli

Christelle Richard,1 Djamel Drider,1 Khalil Elmorjani,2 Didier Marion,2 and Hervé Prévost1*

Laboratoire de Microbiologie Alimentaire et Industrielle, ENITIAA, BP 82225, 44322 Nantes Cedex 3,1 Unité de Recherche sur les Protéines Végétales et leurs Interactions, INRA, BP 7162, 44316 Nantes Cedex 3, France2

Received 30 January 2004/ Accepted 22 March 2004

Divercin V41, a class IIa bacteriocin with strong antilisterial activity, is produced by Carnobacterium divergens V41. To express a recombinant version of divercin V41, we constructed a synthetic gene that encodes the mature divercin V41 peptide and then overexpressed the gene in pET-32b by using the T7 RNA polymerase promoter in the Escherichia coli Origami (DE3)(pLysS) strain. The DvnRV41 peptide was expressed as a translational fusion protein with thioredoxin and accumulated in the cell cytoplasm in a soluble anti-Listeria active form. The fusion protein was then purified and cleaved to obtain pure, soluble, folded DvnRV41 (462 µg per 20 ml of culture). This paper describes the first design of a synthetic bacteriocin gene and the first bacteriocin expressed in the E. coli cytoplasm.

* Corresponding author. Mailing address: Laboratoire de Microbiologie Alimentaire et Industrielle, ENITIAA, Rue de la Géraudière, BP 82225, 44322 Nantes Cedex 3, France. Phone: 33-2-51785524. Fax: 33-2-51785520. E-mail: prevost{at}enitiaa-nantes.fr.

Journal of Bacteriology, July 2004, p. 4276-4284, Vol. 186, No. 13
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.13.4276-4284.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rihakova, J., Petit, V. W., Demnerova, K., Prevost, H., Rebuffat, S., Drider, D. (2009). Insights into Structure-Activity Relationships in the C-Terminal Region of Divercin V41, a Class IIa Bacteriocin with High-Level Antilisterial Activity. Appl. Environ. Microbiol. 75: 1811-1819 [Abstract] [Full Text]  
  • Calvez, S., Rince, A., Auffray, Y., Prevost, H., Drider, D. (2007). Identification of new genes associated with intermediate resistance of Enterococcus faecalis to divercin V41, a pediocin-like bacteriocin. Microbiology 153: 1609-1618 [Abstract] [Full Text]  
  • Drider, D., Fimland, G., Hechard, Y., McMullen, L. M., Prevost, H. (2006). The Continuing Story of Class IIa Bacteriocins. Microbiol. Mol. Biol. Rev. 70: 564-582 [Abstract] [Full Text]  


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