Heterologous expression of enterocin A, a bacteriocin from Enterococcus faecium, fused to a cellulose-binding domain in Escherichia coli results in a functional protein with inhibitory activity against Listeria
Autor(en): | Klocke, M Mundt, K Idler, F Jung, S Backhausen, JE |
Stichwörter: | Biotechnology & Applied Microbiology; CHEESE; FAECALIS; GENETIC-CHARACTERIZATION; L50B; LACTOCOCCUS-LACTIS; MILK; MONOCYTOGENES; PRODUCT | Erscheinungsdatum: | 2005 | Herausgeber: | SPRINGER | Journal: | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | Volumen: | 67 | Ausgabe: | 4 | Startseite: | 532 | Seitenende: | 538 | Zusammenfassung: | The genes for the bacteriocins enterocin A and B were isolated from Enterococcus faecium ATB 197a. Using the pET37b(+) vector, the enterocin genes were fused to an Escherichia coli specific export signal sequence, a cellulose-binding domain (CBDcenA) and a S-tag under the control of a T7lac promotor. The constructs were subsequently cloned into E. coli host cells. The expression of the recombinant enterocins had different effects on both the host cells and other Gram-positive bacteria. The expression of entA in Esc. coli led to the synthesis and secretion of functional active enterocin A fusion proteins, which were active against some Gram-positive indicator bacteria, but did not influence the viability of the host cells. In contrast, the expression of enterocin B fusion proteins led to a reduced viability of the host cells, indicating a misfolding of the protein or interference with the cellular metabolism of Esc. coli. Indicator strains of Gram-positive bacteria were not inhibited by purified enterocin B fusion proteins. However, recombinant enterocin B displayed inhibitory activity after the proteolytic cleavage of the fused peptides. |
ISSN: | 01757598 | DOI: | 10.1007/s00253-004-1838-5 |
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