Edeine, synthesized by Brevibacillus brevis X23, is a linear peptide antibiotic not encoded by ribosomal RNA, renowned for its extensive range of antibacterial effectiveness. Transcriptome sequencing revealed an inverse expression pattern between the two-component regulatory factors yvrG/yvrH and the edeine biosynthesis gene cluster (ede BGC), suggesting the involvement of YvrH/YvrG in the transcriptional regulation of ede BGC. This study utilized Red/ET homologous recombination technology to construct X23 mutant strains that yvrG/yvrH lacks, complemented strains, and overexpressing strains, and investigated the impact of YvrG/YvrH on edeine biosynthesis through phenotype analysis and quantitative real-time PCR (RT-qPCR) techniques. The results revealed that knocking out yvrG/yvrH significantly reduced the antibacterial activity of the strains, the expression of ede BGC decreased, the synthesis of edeine was inhibited, and the yield of edeine was reduced by 46.75%. In summary, YvrG/YvrH act as positive regulatory factors in the edeine biosynthesis. This study provides gene regulatory elements for the metabolic engineering of edeine.