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2014 | 1 | 1 |

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Co-expression and characterization of enterocin CRL35 and its mutant in Escherichia coli Rosetta


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Even though many sequences and structures of bacteriocins from lactic acid bacteria have been fully characterized so far, little information is currently available about bacteriocins heterologously produced by Escherichia coli. For this purpose, the structural gene of enterocin CRL35, munA, was PCR-amplified using specific primers and cloned downstream of PelB sequence in the pET22b (+) expression vector. E. coli Rosetta (DE3) pLysS was chosen as the host for production and enterocin was purified by an easy two-step protocol. The bacteriocin was correctly expressed with the expected intramolecular disulfide bond. Nevertheless, it was found that a variant of the enterocin, differing by 12 Da from the native polypeptide, was co-expressed by E. coli Rosetta in comparable amount. Indeed, the mutant bacteriocin contained two amino acid substitutions that were characterized by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) and HPLCelectrospray (ESI)-Q-TOF tandem mass spectrometry (MS/ MS) sequencing. This is the first report regarding the production of mutants of pediocin-like bacteriocins in the E. coli expression system.








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1 - 1 - 2014
24 - 4 - 2014
26 - 7 - 2013
6 - 12 - 2013


  • Instituto Superior de Investigaciones Biologicas (CONICET-UNT) e Instituto de Quimica Biologica ‘‘Dr. Bernabe Bloj’’, San Miguel de Tucuman, Argentina
  • Istituto di Scienze dell’Alimentazione – Consiglio Nazionale delle Ricerche (CNR), Via Roma, 64 – 83100 Avellino, Italy
  • Instituto Superior de Investigaciones Biologicas (CONICET-UNT) e Instituto de Quimica Biologica ‘‘Dr. Bernabe Bloj’’, San Miguel de Tucuman, Argentina
  • Instituto Superior de Investigaciones Biologicas (CONICET-UNT) e Instituto de Quimica Biologica ‘‘Dr. Bernabe Bloj’’, San Miguel de Tucuman, Argentina
  • Centro de Referencia para Lactobacilos (CERELA-CONICET) S.M. de Tucuman, Argentina
  • Instituto Superior de Investigaciones Biologicas (CONICET-UNT) e Instituto de Quimica Biologica ‘‘Dr. Bernabe Bloj’’, San Miguel de Tucuman, Argentina
  • Centro de Referencia para Lactobacilos (CERELA-CONICET) S.M. de Tucuman, Argentina
  • Chacabuco 461, S.M. de Tucuman (T4000ILI), Argentina, Fax: +54 0381 4248921, Tel: +54 0381 4248921


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