High-level heterologous production and functional expression of the sec-dependent enterocin P from Enterococcus faecium P13 in Lactococcus lactis

Gutiérrez, José Félix Pérez; Larsen, Rasmus; Cintas, Luis M.; Kok, Jan; Hernández, Pablo E.

doi:10.1007/s00253-005-0233-1

Cited by 43 publications

(29 citation statements)

References 66 publications

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“…The choice of LAB and Saccharomyces cerevisiae as heterologous hosts presents a real opportunity for future applications. Within LAB, Lactococcus lactis may offer advantages due to the availability of tools for its genetic manipulation (121), and continuing academic research on this model organism will undoubtedly increase its utility as a heterologous host for the expression of many class IIa bacteriocins, as recently reported for enterocin P (75,86) and divercin V41 (Drider, unpublished results). Finally, most class IIa bacteriocins heterologously produced in LAB have been clearly reported elsewhere (158) and will not be discussed in this review.…”

Section: Purification and Heterologous Production Of Class Iia Bactermentioning

confidence: 99%

The Continuing Story of Class IIa Bacteriocins

Drider

Fimland

Héchard

et al. 2006

Microbiol Mol Biol Rev

646

543

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SUMMARY Many bacteria produce antimicrobial peptides, which are also referred to as peptide bacteriocins. The class IIa bacteriocins, often designated pediocin-like bacteriocins, constitute the most dominant group of antimicrobial peptides produced by lactic acid bacteria. The bacteriocins that belong to this class are structurally related and kill target cells by membrane permeabilization. Despite their structural similarity, class IIa bacteriocins display different target cell specificities. In the search for new antibiotic substances, the class IIa bacteriocins have been identified as promising new candidates and have thus received much attention. They kill some pathogenic bacteria (e.g., Listeria) with high efficiency, and they constitute a good model system for structure-function analyses of antimicrobial peptides in general. This review focuses on class IIa bacteriocins, especially on their structure, function, mode of action, biosynthesis, bacteriocin immunity, and current food applications. The genetics and biosynthesis of class IIa bacteriocins are well understood. The bacteriocins are ribosomally synthesized with an N-terminal leader sequence, which is cleaved off upon secretion. After externalization, the class IIa bacteriocins attach to potential target cells and, through electrostatic and hydrophobic interactions, subsequently permeabilize the cell membrane of sensitive cells. Recent observations suggest that a chiral interaction and possibly the presence of a mannose permease protein on the target cell surface are required for a bacteria to be sensitive to class IIa bacteriocins. There is also substantial evidence that the C-terminal half penetrates into the target cell membrane, and it plays an important role in determining the target cell specificity of these bacteriocins. Immunity proteins protect the bacteriocin producer from the bacteriocin it secretes. The three-dimensional structures of two class IIa immunity proteins have been determined, and it has been shown that the C-terminal halves of these cytosolic four-helix bundle proteins specify which class IIa bacteriocin they protect against.

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Section: Purification and Heterologous Production Of Class Iia Bactermentioning

confidence: 99%

The Continuing Story of Class IIa Bacteriocins

Drider

Fimland

Héchard

et al. 2006

Microbiol Mol Biol Rev

646

543

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“…The production of HirJM79 by all LAB strains was higher than that by E. hirae DCH5 (Table 3), whereas the production of HirJM79 in L. lactis IL1403 was larger than that in L. lactis NZ9000 transformed with the same vectors. Differences in the production of bacteriocins by lactococcal strains have been reported previously (23,28,37,38) and may reflect unknown metabolic differences between L. lactis subsp. lactis IL1403 and L. lactis subsp.…”

Section: Discussionmentioning

confidence: 97%

“…Indeed, bacteriocin producers are protected from their own bacteriocin by the concomitant expression of a cognate immunity protein. These proteins act either by affecting bacteriocin aggregation and pore formation or by disturbing the interaction between the bacteriocin and the membrane-located bacteriocin receptor (23). Coexpression of the structural and immunity genes also increased the heterologous production of enterocin P (EntP), a Sec-dependent bacteriocin produced by E. faecium P13 in lactococci (23).…”

Section: Discussionmentioning

confidence: 99%

“…All chromatographic columns and supporting gels were from Amersham Biosciences Europe GmbH (Cerdanyola, Spain). Purified fractions from the last RP-FPLC step were subjected to matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, as described previously (23).…”

Section: Methodsmentioning

confidence: 99%

“…However, since enterococci are involved in food spoilage, nosocomial infections, and the spread of antibiotic resistance (36,41), interest in the heterologous production and functional expression of enterocins in other microbial hosts is growing rapidly. Moreover, the production of enterocins in other hosts could lead to increased bacteriocin production, production of bacteriocins in safer hosts, obtention of multibacteriocinogenic strains with wider antimicrobial spectra, and the transfer of antimicrobial capabilities to LAB that are useful as starter, protective, and probiotic cultures in food (23).…”

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confidence: 99%

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Cloning and Heterologous Production of Hiracin JM79, a Sec-Dependent Bacteriocin Produced by Enterococcus hirae DCH5, in Lactic Acid Bacteria and Pichia pastoris

Sánchez

Borrero

Gómez‐Sala

et al. 2008

Appl Environ Microbiol

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Hiracin JM79 (HirJM79), a Sec-dependent bacteriocin produced by Enterococcus hirae DCH5, was cloned and produced in Lactococcus lactis, Lactobacillus sakei, Enterococcus faecium, Enterococcus faecalis, and Pichia pastoris. For heterologous production of HirJM79 in lactic acid bacteria (LAB), the HirJM79 structural gene (hirJM79), with or without the HirJM79 immunity gene (hiriJM79), was cloned into the plasmid pMG36c under the control of the constitutive promoter P 32 and into the plasmid pNZ8048 under the control of the inducible P NisA promoter. For the production of HirJM79 in P. pastoris, the gene encoding the mature HirJM79 protein was cloned into the pPICZ␣A expression vector. The recombinant plasmids permitted the production of biologically active HirJM79 in the supernatants of L. lactis IL1403, L. lactis NZ9000, L. sakei Lb790, E. faecalis JH2-2, and P. pastoris X-33, the coproduction of HirJM79 and nisin A in L. lactis DPC5598, and the coproduction of HirJM79 and enterocin P in E. faecium L50/14-2. All recombinant LAB produced larger quantities of HirJM79 than E. hirae DCH5, although the antimicrobial activities of most transformants were lower than that predicted from their production of HirJM79. The synthesis, processing, and secretion of HirJM79 proceed efficiently in recombinant LAB strains and P. pastoris.

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