1887

Abstract

Bovicin HJ50 is a new lantibiotic containing a disulfide bridge produced by HJ50; its encoding gene was reported in our previous publication. To identify other genes involved in bovicin HJ50 production, DNA fragments flanking were cloned and sequenced. The bovicin HJ50 biosynthesis gene locus was encoded by a 9.9 kb region of chromosomal DNA and consisted of at least nine genes in the following order: , -, -, -, - ORF1, ORF2, and . A thiol–disulfide oxidoreductase gene named was located downstream of . A knockout mutant of this gene retained antimicrobial activity and the molecular mass of bovicin HJ50 in the mutant was the same as that of bovicin HJ50 in HJ50, implying that is not involved in bovicin HJ50 production. Transcriptional analyses showed that and constituted an operon, and the transcription start site of the promoter was located at a G residue 45 bp upstream of the translation start codon for , while through were transcribed together and the transcription start site of the promoter was located at a C residue 35 bp upstream of . We also demonstrated successful heterologous expression of bovicin HJ50 in MG1363, which lacks thiol–disulfide oxidoreductase genes; this showed that thiol–disulfide oxidoreductase genes other than are not essential for bovicin HJ50 biosynthesis.

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2009-02-01
2024-03-29
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