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Appl Environ Microbiol

Title:		"Use of the yeast Pichia pastoris as an expression host for secretion of enterocin L50, a leaderless two-peptide (L50A and L50B) bacteriocin from Enterococcus faecium L50"
Author(s):		Basanta A; Gomez-Sala B; Sanchez J; Diep DB; Herranz C; Hernandez PE; Cintas LM;
Address:		"Departamento de Nutricion, Bromatologia y Tecnologia de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain"
Journal Title:		Appl Environ Microbiol
Year:		2010
Volume:		20100326
Issue:		10
Page Number:		3314 - 3324
DOI:		10.1128/AEM.02206-09
ISSN/ISBN:		1098-5336 (Electronic) 0099-2240 (Print) 0099-2240 (Linking)
Abstract:		"In this work, we report the expression and secretion of the leaderless two-peptide (EntL50A and EntL50B) bacteriocin enterocin L50 from Enterococcus faecium L50 by the methylotrophic yeast Pichia pastoris X-33. The bacteriocin structural genes entL50A and entL50B were fused to the Saccharomyces cerevisiae gene region encoding the mating pheromone alpha-factor 1 secretion signal (MFalpha1(s)) and cloned, separately and together (entL50AB), into the P. pastoris expression and secretion vector pPICZalphaA, which contains the methanol-inducible alcohol oxidase promoter (P(AOX1)) to express the fusion genes. After transfer into the yeast, the recombinant plasmids were integrated into the genome, resulting in three bacteriocinogenic yeast strains able to produce and secrete the individual bacteriocin peptides EntL50A and EntL50B separately and together. The secretion was efficiently directed by MFalpha1(s) through the Sec system, and the precursor peptides were found to be correctly processed to form mature and active bacteriocin peptides. The present work describes for the first time the heterologous expression and secretion of a two-peptide non-pediocin-like bacteriocin by a yeast"
Keywords:		Anti-Bacterial Agents/chemistry/metabolism/pharmacology Bacteriocins/chemistry/*genetics/isolation & purification/*metabolism Enterococcus faecium/*genetics Genetic Vectors/genetics Pediococcus/drug effects Pichia/*genetics/*metabolism Plasmids Recombinan;
Notes:		"MedlineBasanta, Antonio Gomez-Sala, Beatriz Sanchez, Jorge Diep, Dzung B Herranz, Carmen Hernandez, Pablo E Cintas, Luis M eng Research Support, Non-U.S. Gov't 2010/03/30 Appl Environ Microbiol. 2010 May; 76(10):3314-24. doi: 10.1128/AEM.02206-09. Epub 2010 Mar 26"