Transfer of nisin gene cluster from Lactococcus lactis ATCC 11454 into the chromosome of Bacillus subtilis 168

Abstract

Nisin is an antimicrobial peptide produced by certain strains of Lactococcus lactis. It is a gene-encoded peptide that contains unusual amino acid residues. These novel residues are introduced by posttranslational modification machinery and confer unique chemical and physical properties that are not attainable by regular amino acid residues. To study the modification mechanisms and to create structural analogs with superior properties, it would be advantageous to insert the nisin genes into a bacterial strain that is amenable to genetic manipulation. In this study, we report the cloning and integration of the complete and intact nisin gene cluster into the Bacillus subtilis 168 chromosome. Furthermore, we demonstrate that the nisin genes are transcriptionally active. These results should greatly facilitate the studies of the genes and proteins involved in nisin expression, as well as provide a standard system for the manipulation and expression of genes involved in other members of the lantibiotic family of antimicrobial peptides.

Biosynthesis, immunity, regulation, mode of action and engineering of the model lantibiotic nisin

“…Alternatively, heterologous expression may raise the yield of nisin variants and/or facilitate engineering. In this respect it is interesting to note that the nisin gene cluster has been successfully inserted in the chromosome of Bacillus subtilis 168 [98]. Generally, it is not recommended to use the same compound for both food conservation and for antibiotic treatment.…”

Expression of the Lantibiotic Mersacidin in Bacillus amyloliquefaciens FZB42

“…Successful heterologous production has previously been shown for several lantibiotics, e. g. subtilin and nisin by B. subtilis 168 [8], [9], lacticin 3147 by Enterococcus faecalis [10], or epicidin 280 by Staphylococcus carnosus [11]. Very recently, production of active lichenicidin has even been achieved in Escherichia coli [12].…”

Simple and rapid direct cloning and heterologous expression of natural product biosynthetic gene cluster in Bacillus subtilis via Red/ET recombineering

“…Third, B. subtilis has sophisticated and simple genetic operation approaches and tools, particularly for the integration of large foreign DNA fragments into the chromosome by natural transformation, with a capacity of more than 3 Mb 17 18 . Several secondary metabolite biosynthetic pathways have been successfully expressed in B. subtilis , for example, bacitracin biosynthetic gene cluster from B. licheniformis 19 , nisin biosynthetic gene cluster from Lactococcus lactis 20 , polymyxin biosynthetic gene cluster from Paenibacillus polymyxa 21 , enniatin biosynthetic gene cluster from Fusarium .sp 22 , amicoumacin biosynthetic gene cluster from B. subtilis 14 , and 6 -deoxyerythronolide B biosynthetic gene cluster from Saccharopolyspora erythraea 23 .…”