摘　要：伊短菌素（edeine）是短短芽孢杆菌（Brevibacillus brevis）X23产生的线性非核糖体抗菌肽，具有抑菌谱广、抑菌活性强的特点。通过转录组测序，发现双组分调控因子yvrG/yvrH与伊短菌素生物合成基因簇（ede BGC）的表达模式相反，推测YvrG/YvrH可能参与ede BGC的转录调控。本研究利用Red/ET同源重组技术构建yvrG/yvrH的X23缺失菌株、回补菌株、过表达菌株，通过表型分析及实时荧光定量PCR（RT-qPCR）技术研究YvrG/YvrH对伊短菌素生物合成的影响。结果显示，ΔyvrG/yvrH的抑菌活性显著降低，且ede BGC的表达水平下调，伊短菌素的合成受到了抑制，产量降低了46.75%。综上所述，YvrG/YvrH正调控伊短菌素的生物合成。本研究为伊短菌素代谢工程改造提供了基因调控元件。
关键词：短短芽孢杆菌；伊短菌素；双组分调控因子；抑菌活性；生物合成
中图分类号：S476+.8                         文献标志码：ADOI：10.3969/j.issn.1007-7146.2024.05.008

Abstract: 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.
Key words: Brevibacillus brevis; edeine; two-component regulatory factors; bacteriostatic activity; biosynthesis
（Acta Laser Biology Sinica, 2024, 33(5): 461-469）