期刊
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
卷 40, 期 9, 页码 977-993出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10295-013-1302-6
关键词
Bacteriocins; Sakacin A; Sakacin A-derived chimeras; Lactic acid bacteria (LAB); Pichia pastoris; Kluyveromyces lactis; Heterologous bacteriocin production
资金
- Ministerio de Economia y Competitividad (MINECO) [AGL2012-34829]
- Ministerio de Ciencia e Innovacion (MICINN) [AGL2009-08348]
- BSCH-UCM [GR35-10A]
- Comunidad de Madrid (CAM) [S2009/AGR-1489]
- Ministerio de Educacion y Ciencia (MEC), Spain
Mature sakacin A (SakA, encoded by sapA) and its cognate immunity protein (SakI, encoded by sapiA), and two SakA-derived chimeras mimicking the N-terminal end of mature enterocin P (EntP/SakA) and mature enterocin A (EntA/SakA) together with SakI, were fused to different signal peptides (SP) and cloned into the protein expression vectors pNZ8048 and pMG36c for evaluation of their production and functional expression by different lactic acid bacteria. The amount, antimicrobial activity, and specific antimicrobial activity of SakA and its chimeras produced by Lactococcus lactis subsp. cremoris NZ9000 depended on the SP and the expression vector. Only L. lactis NZ9000 (pNUPS), producing EntP/SakA, showed higher bacteriocin production and antimicrobial activity than the natural SakA-producer Lactobacillus sakei Lb706. The lower antimicrobial activity of the SakA-producer L. lactis NZ9000 (pNUS) and that of the EntA/SakA-producer L. lactis NZ9000 (pNUAS) could be ascribed to secretion of truncated bacteriocins. On the other hand, of the Lb. sakei Lb706 cultures transformed with the pMG36c-derived vectors only Lb. sakei Lb706 (pGUS) overproducing SakA showed a higher antimicrobial activity than Lb. sakei Lb706. Finally, cloning of SakA and EntP/SakA into pPICZ alpha A and pKLAC2 permitted the production of SakA and EntP/SakA by recombinant Pichia pastoris X-33 and Kluyveromyces lactis GG799 derivatives although their antimicrobial activity was lower than expected from their production.
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