Development of bacteriocinogenic strains of Saccharomyces cerevisiae heterologously expressing and secreting the leaderless enterocin L50 peptides L50A and L50B from Enterococcus faecium L50.
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 (MFalpha1(s)) 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 P(GAL1). The generation of recombinant S. cerevisiae strains heterologously expressing and secreting biologically active EntL50A and EntL50B demonstrates the suitability of the MFalpha1(s)-containing vector pYABD01 to direct processing and secretion of these antimicrobial peptides through the S. cerevisiae Sec system.
Basanta
Antonio
A
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Herranz
Carmen
C
Gutiérrez
Jorge
J
Criado
Raquel
R
Hernández
Pablo E
PE
Cintas
Luis M
LM
D000890:Anti-Infective Agents / Q000378:metabolism* / Q000494:pharmacology; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D016984:Enterococcus faecium / Q000235:genetics*; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D010957:Plasmids; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D000890:Anti-Infective Agents; D001430:Bacteriocins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C111937:enterocin L50A; C111938:enterocin L50B