全局调控因子ccpA调控短短芽胞杆菌X23伊短菌素生物合成的研究

doi:10.16409/j.cnki.2095-039x.2025.02.036

中国生物防治学报 ›› 2025, Vol. 41 ›› Issue (4): 826-835.DOI: 10.16409/j.cnki.2095-039x.2025.02.036

• 研究论文 • 上一篇

全局调控因子ccpA调控短短芽胞杆菌X23伊短菌素生物合成的研究

陈子玥¹, 付竞¹, 张亮¹, 蓝若伊¹, 刘天波², 刘清术³, 王运生¹, 陈武¹

1. 湖南农业大学植物保护学院, 长沙 410128;
2. 湖南省烟草科学研究所, 长沙 410004;
3. 湖南省微生物研究院, 长沙 410009

收稿日期:2025-01-12 发布日期:2025-08-15
通讯作者: 陈武
作者简介:陈子玥，女，硕士研究生，E-mail： chenziyue0644@163.com；通信作者，陈武，博士，副教授，E-mail： chenwuwarrior@163.com。
基金资助:
国家自然科学基金（32472641）；国家烟草专卖局科技项目（110220001019）

Regulation of Global Regulator ccpA on Biosynthesis of Ibremycin in Brevibacillus brevis X23

CHEN Ziyue¹, FU Jing¹, ZHANG Liang¹, LAN Ruoyi¹, LIU Tianbo², LIU Qingshu³, WANG Yunsheng¹, CHEN Wu¹

1. College of Plant Protection, Hunan Agricultural University, Changsha 410128, China;
2. Hunan Tobacco Science Research Institute, Changsha 410004, China;
3. Hunan Provincial Microbiology Research Institute, Changsha 410009, China

Received:2025-01-12 Published:2025-08-15

摘要/Abstract

摘要： 分解代谢物控制蛋白A（CcpA）参与协调碳代谢，是革兰氏阳性菌的关键调节因子之一。已有研究揭示了CcpA在碳代谢中的重要作用，但其在调控伊短菌素生物合成中的作用鲜见报道。伊短菌素的合成需要消耗大量的能量和前体物质，而碳源作为提供能量和碳骨架的关键，其代谢受到CcpA的调控。本研究利用Red/ET同源重组法敲除野生型短短芽胞杆菌Brevibacillus brevis X23中的ccpA基因，获得突变体菌株X23Δ ccpA。经平皿对峙培养、qRT-PCR和HPLC-MS定量分析，发现X23ΔccpA的抑菌圈直径较X23-WT显著增大，伊短菌素生物合成基因簇中edeP表达量显著上调、伊短菌素的峰面积提高了44.87%，表明CcpA是伊短菌素生物合成的负转录调控因子。本研究为伊短菌素的代谢工程改造提供了候选转录调控因子元件。

关键词: 伊短菌素, 分解代谢物控制蛋白A, 短短芽胞杆菌X23, 碳代谢

Abstract: The catabolite control protein A (CcpA) is involved in the coordination of carbon metabolism and is one of the key regulators of Gram-positive bacteria. Although studies have revealed the important role of CcpA in carbon metabolism, its role in regulating edeine biosynthesis is rarely reported. The synthesis of edeine requires a large amount of energy and precursor substances, and carbon source is the key to providing energy and carbon skeleton, and its metabolism is regulated by CcpA. In this study, the ccpA in wild-type Brevibacillus brevis X23 (X23-WT) strain was knocked out by Red/ET homologous recombination method, and the mutant strain X23ΔccpA was obtained. By pair-culturing assay, qRT-PCR and HPLC-MS quantitative analysis, it was found that the inhibition zone diameter of X23ΔccpA was significantly bigger than that of X23-WT, the expression of edeP in edeine biosynthetic gene cluster was significantly up-regulated, and the peak area of edeine was increased by 44.87%, indicating that CcpA is a negative transcriptional regulator of edeine biosynthesis. This study provides candidate transcriptional regulatory factor elements for the metabolic engineering of edeine.

Key words: edeine, metabolite control protein A, Brevibacillus brevis X23, carbon metabolism

中图分类号:

S476

陈子玥, 付竞, 张亮, 蓝若伊, 刘天波, 刘清术, 王运生, 陈武. 全局调控因子ccpA调控短短芽胞杆菌X23伊短菌素生物合成的研究[J]. 中国生物防治学报, 2025, 41(4): 826-835.

CHEN Ziyue, FU Jing, ZHANG Liang, LAN Ruoyi, LIU Tianbo, LIU Qingshu, WANG Yunsheng, CHEN Wu. Regulation of Global Regulator ccpA on Biosynthesis of Ibremycin in Brevibacillus brevis X23[J]. Chinese Journal of Biological Control, 2025, 41(4): 826-835.

参考文献

[1] Qiu S, Yang H, Zhao C, et al. The identification and antibacterial activity research of an endogenous Brevibacillus brevis[J]. Natural Product Research and Development, 2009, 21: 30-33.
[2] Hshida O, Takagi H, Kadowaki K, et al. Proposal for two new genera, Brevibacillus gen. nov. and Aneurinibacillus gen. nov.[J]. International Journal of Systematic Bacteriology, 1996, 46(4): 939-46.
[3] Kurylo-borowska Z. Isolation and properties of pure edeine, an antibiotic of the strain[J]. Biuletyn Instytutu Medycyny Morskiej Gdansku, 1959,10: 151-63.
[4] Jie D, Cuiyang Z, Qingshan L, et al. Characterization of a pathway-specific activator of edeine biosynthesis and improved edeine production by its overexpression in Brevibacillus brevis[J]. Frontiers in Plant Science, 2022, 13: 1022476.
[5] Yan Z, Guanze L, Feng Y, et al. Multilayered regulation of secondary metabolism in medicinal plants[J]. Molecular Horticulture, 2023, 3(1): 11.
[6] Chen W, Wang Y, Li D, et al. Draft genome sequence of Brevibacillus brevis strain X23, a biocontrol agent against bacterial wilt[J]. Journal of Bacteriology, 2012, 194(23): 6634-6635.
[7] Liu Q, Shen Q, Bian X, et al. Simple and rapid direct cloning and heterologous expression of natural product biosynthetic gene cluster in Bacillus subtilis via Red/ET recombineering[J]. Scientific Reports, 2016, 6: 34623.
[8] Liu Q, Zhang L, Wang Y, et al. Enhancement of edeine production in Brevibacillus brevis X23 via in situ promoter engineering[J]. Microbial Biotechnology, 2022, 15(2): 577-89.
[9] 邱丽婷, 张亮, 刘天波, 等. 双组分因子LiaRS调控短短芽胞杆菌X23伊短菌素生物合成的研究[J]. 中国生物防治学报, 2024, 40(5): 1036-1044.
[10] 于佳, 张亮, 刘天波, 等. 双组分系统YvrG/YvrH调控伊短菌素生物合成的研究[J]. 激光生物学报, 2024, 33(5): 461-469.
[11] El-banna N M. Effect of carbon source on the antimicrobial activity of the air flora[J]. World Journal of Microbiology and Biotechnology, 2005, 21: 1451-1454.
[12] Zhang Y, Xiao F, Zhang L, et al. A new mechanism of carbon metabolism and acetic acid balance regulated by ccpa[J]. Microorganisms, 2023, 11(9): 2303.
[13] Sadykov M R, Hartmann T, Mattes T A, et al. CcpA coordinates central metabolism and biofilm formation in Staphylococcus epidermidis[J]. Microbiology, 2011, 157(12): 3458-3468.
[14] Zheng L, Chen Z, Itzek A, et al. CcpA regulates biofilm formation and competence in Streptococcus gordonii[J]. Molecular Oral Microbiology, 2012, 27(2): 83-94.
[15] Fujita Y, Miwa Y, Galinier A, et al. Specific recognition of the Bacillus subtilis gnt cis-acting catabolite-responsive element by a protein complex formed between CcpA and seryl-phosphorylated HPr[J]. Molecular microbiology, 1995, 17(5): 953-960.
[16] Jones B E, Dossonnet V, Kuster E, et al. Binding of the catabolite repressor protein CcpA to its DNA target is regulated by phosphorylation of its corepressor HPr[J]. Journal of Biological Chemistry, 1997, 272(42): 26530-26535.
[17] Lorca G L, Chung Y J, Barabote R D, et al. Catabolite repression and activation in Bacillus subtilis: dependency on CcpA, HPr, and HprK[J]. Journal of bacteriology, 2005, 187(22): 7826-7839.
[18] Chauvaux S. CcpA and HPr (ser-P): mediators of catabolite repression in Bacillus subtilis[J]. Research in microbiology, 1996, 147(6-7): 518-522.
[19] Grundy F J, Turinsky A J, Henkin T M. Catabolite regulation of Bacillus subtilis acetate and acetoin utilization genes by ccpA[J]. Journal of Bacteriology, 1994, 176(15): 4527-33.
[20] Wright Mitchell H, Adelskov J, Greene Anthony C. Bacterial DNA extraction using individual enzymes and phenol/chloroform separation[J]. Journal of Microbiology &, Biology Education, 2017, 18(2): 101128.
[21] Takahashi W, Yamagata H, Yamaguchi K, et al. Genetic transformation of Bacillus brevis 47, a protein-secreting bacterium, by plasmid DNA[J]. Journal of Bacteriology, 1983, 156(3): 1130-1134.
[22] Liang Z, Cui L, Duan C, et al. Effect of transcription factor AbrB on the biosynthesis of antibiotic edeines in biocontrol bacteria Brevibacillus brevis X23[J]. Journal of Biological Control, 2020, 36: 564.
[23] 张亮, 吕翠, 段彩琛, 等. 转录调控因子AbrB对生防短短芽胞杆菌X23中抗生素edeines生物合成的影响[J]. 中国生物防治学报, 2020, 36(4): 564-574.
[24] Oakley C E, Ahuja M, Sun W W, et al. Discovery of McrA, a master regulator of aspergillus secondary metabolism[J]. Molecular Microbiology, 2017, 103(2): 347-365.

全局调控因子ccpA调控短短芽胞杆菌X23伊短菌素生物合成的研究

Regulation of Global Regulator ccpA on Biosynthesis of Ibremycin in Brevibacillus brevis X23

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 1

编辑推荐

Metrics

本文评价

联系我们