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Applied and Environmental Microbiology, April 2009, p. 2382-2392, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.01476-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development of Bacteriocinogenic Strains of Saccharomyces cerevisiae Heterologously Expressing and Secreting the Leaderless Enterocin L50 Peptides L50A and L50B from Enterococcus faecium L50{triangledown}

Antonio Basanta, Carmen Herranz, Jorge Gutiérrez, Raquel Criado, Pablo E. Hernández, and Luis M. Cintas*

Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain

Received 1 July 2008/ Accepted 2 February 2009

A segregationally stable expression and secretion vector for Saccharomyces cerevisiae, named pYABD01, was constructed by cloning the yeast gene region encoding the mating pheromone {alpha}-factor 1 secretion signal (MF{alpha}1s) into the S. cerevisiae high-copy-number expression vector pYES2. The structural genes of the two leaderless peptides of enterocin L50 (EntL50A and EntL50B) from Enterococcus faecium L50 were cloned, separately (entL50A or entL50B) and together (entL50AB), into pYABD01 under the control of the galactose-inducible promoter PGAL1. The generation of recombinant S. cerevisiae strains heterologously expressing and secreting biologically active EntL50A and EntL50B demonstrates the suitability of the MF{alpha}1s-containing vector pYABD01 to direct processing and secretion of these antimicrobial peptides through the S. cerevisiae Sec system.

* Corresponding author. Mailing address: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain. Phone: 34-913943751. Fax: 34-913943743. E-mail: lcintas{at}vet.ucm.es

{triangledown} Published ahead of print on 13 February 2009.

Applied and Environmental Microbiology, April 2009, p. 2382-2392, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.01476-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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