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

doi:10.16409/j.cnki.2095-039x.2024.02.056

中国生物防治学报 ›› 2024, Vol. 40 ›› Issue (5): 1036-1044.DOI: 10.16409/j.cnki.2095-039x.2024.02.056

• 研究论文 • 上一篇

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

邱丽婷¹, 张亮², 刘天波³, 巢进⁴, 蔡海林⁵, 易克⁶, 刘清术⁷, 陈武¹

1. 湖南农业大学植物保护学院, 长沙 410128;
2. 湖南农业大学农学院, 长沙 410128;
3. 湖南省烟草科学研究所, 长沙 410004;
4. 湖南省湘西自治州烟草公司技术中心, 湘西 416000;
5. 湖南省长沙市烟草公司技术中心, 长沙 410019;
6. 湖南中烟工业有限责任公司原料采购中心, 长沙 410007;
7. 湖南省微生物研究院, 长沙 410009

收稿日期:2024-04-01 发布日期:2024-10-11
作者简介:邱丽婷，女，硕士研究生，E-mail:1290560702@qq.com;通信作者，陈武，男，博士，副教授，E-mail:wuchen77@hunau.edu.cn。
基金资助:
国家自然科学基金（32472641）；中国烟草总公司科技项目（110220001019）；湖南省技术攻关“揭榜挂帅”（2021NK1040）；湖南中烟工业有限责任公司科技项目（KY2023YC0009）；长沙市烟草公司科技项目（CS2022KJ02）

Effects of Two-Component System LiaRS on Biosynthesis of Edeine from Brevibacillus brevis X23

QIU Liting¹, ZHANG Liang², LIU Tianbo³, CHAO Jin⁴, CAI Hailin⁵, YI Ke⁶, LIU Qingshu⁷, CHEN Wu¹

1. College of Plant Protection, Hunan Agricultural University, Changsha 410128, China;
2. College of Agronomy, Hunan Agricultural University, Changsha 410128, China;
3. Hunan Tobacco Science Research Institute, Changsha 410004, China;
4. Hunan Xiangxi autonomous prefecture tobacco company technology center, Xiangxi 416000, China;
5. Hunan Provincial Tobacco Company Technology Center, Changsha 410019, China;
6. Hunan Tobacco Industry Co., Ltd. Raw Material Procurement Center, Changsha 410007, China;
7. Hunan Provincial Microbiology Research Institute, Changsha 410009, China

Received:2024-04-01 Published:2024-10-11

摘要/Abstract

摘要： 双组分系统（two-component system，TCS）参与多种芽胞杆菌次级代谢产物生物合成的调控，但调控伊短菌素生物合成的TCS鲜见报道。前期转录组测序分析发现，在短短芽胞杆菌X23中，双组分因子LiaRS与伊短菌素生物合成基因簇（ede BGC）的表达趋势一致，推测其可能参与伊短菌素生物合成的调控，通过Red/ET同源重组法敲除野生型X23的liaS和liaR基因，构建了突变体菌株，平皿对峙培养、qRT-PCR定量分析和HPLC-MS靶向代谢分析等结果表明，在相同培养条件下，突变菌株的抑菌圈直径较野生型显著增加，伊短菌素生物合成基因簇的表达量显著上调，且伊短菌素的峰面积是野生型的1.3倍。本研究发现LiaRS是伊短菌素生物合成的负调控因子，为伊短菌素的代谢工程改造提供了候选基因元件。

关键词: 短短芽胞杆菌, 伊短菌素, 双组分系统, LiaRS, Red/ET同源重组

Abstract: Two-component systems (TCS) perform a regulatory function in the biosynthesis of various secondary metabolites in Bacillus. However, the TCS controlling edeine biosynthesis have seldom been reported. This research, employing bioinformatics analysis, has uncovered the potential regulatory role of the two-component factor LiaRS in the biosynthesis of edeine. By employing the Red/ET homologous recombination technique, the liaS and liaR genes of the wild-type X23 were effectively eliminated, leading to the development of mutant strains. The results from plate culture, qRT-PCR quantitative analysis, and HPLC-MS targeted metabolic profiling demonstrated a significant increase in the diameter of the inhibitory zone for the mutant strains and a notable up-regulation of the expression of the edeine biosynthesis gene clusters. Moreover, the peak area of edeine was found to be 1.3 times that of the wild type. The findings of this research indicate that LiaRS functions as a suppressor of edeine biosynthesis, thereby offering potential gene elements for the metabolic engineering of edeine.

Key words: Brevibacillus brevis, edeine, two-component system, LiaRS, Red/ET recombination

中图分类号:

S476

邱丽婷, 张亮, 刘天波, 巢进, 蔡海林, 易克, 刘清术, 陈武. 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.

QIU Liting, ZHANG Liang, LIU Tianbo, CHAO Jin, CAI Hailin, YI Ke, LIU Qingshu, CHEN Wu. Effects of Two-Component System LiaRS on Biosynthesis of Edeine from Brevibacillus brevis X23[J]. Chinese Journal of Biological Control, 2024, 40(5): 1036-1044.

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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

Effects of Two-Component System LiaRS on Biosynthesis of Edeine from Brevibacillus brevis X23

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