| Author/Editor | Morisset, Dany; Frere, Jacques | |
| Title | Heterologous expression of bacteriocins using the mesentericin Y105 dedicated transport system by Leuconostoc mesenteroides | |
| Type | članek | |
| Source | Biochimie | |
| Vol. and No. | Letnik 84 | |
| Publication year | 2002 | |
| Volume | str. 569-76 | |
| Language | eng | |
| Abstract | Mesentericin Y105 (MesY105) is a class IIa anti-Listeria bacteriocin, produced by Leuconostoc (Ln.) mesenteroides Y105 and with potential food grads application. This bacterium produces a second bacteriocin, mesentericin B 105 (MesB105), that does not belong to the same class. To study secretion of bacteriocins by the use of the MesY 105 dedicated transport system (DTS), plasmids were constructed for heterologous expression by Ln. mesenteroides. pFBYC04 (Microbiology 144 (1998) 2845) harbours two divergent operons required for McsY105 secretion, i.e. the mesY1 operon, encoding pre-MesY105 and immunity, respectively, and the mesCDE operon for secretion. A pFBYC04 derivative, pDMJF01 was constructed by divergent PCR to remove the mesY gene. Ln. mesenleroides DSM20484(pDMJF01) was unable to produce MesY105. The mesY1 operon and mesB, mesH and mesF genes, encoding pre-MesB105, MesB105 immunity and a putative protein with unknown function, respectively, were cloned independently into a compatible pDMJF01 plasmid to produce, respectively, pDMJF:YI and pDMJF:BHF. DSM20484 transformed independently with these plasmids was unable to secrete any bacteriocin. MesY105 and MesB105 secretion was observed (br DSM20484(pDMJF01) harbouring both pDMJF:YI and pDMJF:BHF. This indicates that the MesY105 DTS permits the transport of MesB105. MesY105 secretion machinery was used to secrete pediocin PA-1 (PcdPA-1) by DSM20484 by an in-frame gene fusion strategy where the gene portions corresponding to the MesY105 leader peptide and the mature PedPA-1 were ligated. Thus, MesY105 secretion machinery appears to be a useful tool for secretion of class II bacteriocins by Leuconostoc. | |
| Descriptors | LEUCONOSTOC BACTERIOCINS PLASMIDS CLONING, MOLECULAR POLYMERASE CHAIN REACTION |