RT Journal Article
T1 Use of Synthetic Genes for Cloning, Production and Functional Expression of the Bacteriocins Enterocin A and Bacteriocin E 50-52 by Pichia pastoris and Kluyveromyces lactis
A1 Jiménez Martínez, Juan J.
A1 Borrero Del Pino, Juan
A1 Gútiez Sainz-Pardo, Loreto
A1 Arbulu Ruiz, Sara
A1 Cintas Izarra, Luis Miguel
A1 Herranz Sorribes, Carmen
A1 Hernández Cruza, Pablo Elpidio
AB The use of synthetic genes may constitute a successful approach for the heterologous production and functional expression of bacterial antimicrobial peptides (bacteriocins) by recombinant yeasts. In this work, synthetic genes with adapted codon usage designed from the mature amino acid sequence of the bacteriocin enterocin A (EntA), produced by Enterococcus faecium T136, and the mature bacteriocin E 50-52 (BacE50-52), produced by E. faecium NRRL B-32746, were synthesized. The synthetic entA and bacE50-52 were cloned into the protein expression vectors pPICZαA and pKLAC2 for transformation of derived vectors into Pichia pastoris X-33 and Kluyveromyces lactis GG799, respectively. The recombinant vectors were linearized and transformed into competent cells selecting for P. pastoris X-33EAS (entA), P. pastoris X-33BE50-52S (bacE50-52), K. lactis GG799EAS (entA), and K. lactis GG799BE50-52S (bacE50-52). P. pastoris X-33EAS and K. lactis GG799EAS, but not P. pastoris X-33BE50-52S and K. lactis GG799BE50-52S, showed antimicrobial activity in their supernatants. However, purification of the supernatants of the producer yeasts permitted recovery of the bacteriocins EntA and BacE50-52. Both purified bacteriocins were active against Gram-positive bacteria such as Listeria monocytogenes but not against Gram-negative bacteria, including Campylobacter jejuni.
PB Springer
SN 1073-6085
YR 2014
FD 2014
LK https://hdl.handle.net/20.500.14352/116681
UL https://hdl.handle.net/20.500.14352/116681
LA eng
NO Jiménez, J. J., Borrero, J., Gútiez, L., Arbulu, S., Herranz, C., Cintas, L. M., & Hernández, P. E. (2014). Use of synthetic genes for cloning, production and functional expression of the bacteriocins enterocin A and bacteriocin E 50-52 by Pichia pastoris and Kluyveromyces lactis. Molecular Biotechnology, 56(6), 571-583. https://doi.org/10.1007/S12033-014-9731-7
NO Ministerio de Economía y Competitividad (España)
NO Ministerio de Ciencia e Innovación (España)
NO Banco de Santander Central Hispano
NO Comunidad de Madrid
NO Universidad Complutense de Madrid
DS Docta Complutense
RD 16 abr 2025