| 解淀粉芽胞杆菌HZ-12中ysnE参与吲哚-3-乙酸合成的代谢途径研究 |
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| 引用本文： | 周慧娟,佘梦林,李孜燕,高林,高加明,蔡冬波,杨之帆,陈守文,马昕.解淀粉芽胞杆菌HZ-12中ysnE参与吲哚-3-乙酸合成的代谢途径研究[J].微生物学报,2023,63(2):845-854. |
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| 作者姓名： | 周慧娟 佘梦林 李孜燕 高林 高加明 蔡冬波 杨之帆 陈守文 马昕 |
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| 作者单位： | 湖北大学生命科学学院 省部共建生物催化与酶工程国家重点实验室 湖北省环境微生物工程技术研究中心, 湖北 武汉 430062;中国农业科学院烟草研究所, 山东 青岛 266101;中国烟草总公司湖北省公司, 湖北 武汉 430000 |
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| 基金项目： | 武汉市知识创新专项-曙光计划(2022020801020334)；湖北省烟草公司科技项目(027Y2020-013) |
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| 摘 要： | 【目的】吲哚-3-乙酸是调控植物生长发育和生理活动的重要激素，吲哚-3-乙酸N-乙酰转移酶YsnE在吲哚-3-乙酸合成中发挥重要作用，本研究拟解析解淀粉芽胞杆菌中YsnE参与吲哚-3-乙酸合成的代谢途径。【方法】通过基因ysnE缺失和强化表达，分析ysnE对吲哚-3-乙酸合成影响，结合吲哚-3-乙酸合成中间物(吲哚丙酮酸、吲哚乙酰胺、色胺和吲哚乙腈)添加和体外酶转化实验，解析ysnE参与吲哚-3-乙酸合成的代谢途径。【结果】明确了YsnE在解淀粉芽胞杆菌HZ-12吲哚-3-乙酸合成中发挥重要作用。发现ysnE缺失菌株中的吲哚丙酮酸、吲哚乙酰胺和吲哚乙腈利用显著降低，揭示了YsnE主要发挥吲哚丙酮酸脱羧酶YclB和吲哚乙酰胺水解酶/腈水解酶/腈水合酶YhcX的功能，并通过参与吲哚丙酮酸、吲哚乙酰胺和吲哚乙腈途径来影响吲哚-3-乙酸合成。【结论】初步揭示了YsnE通过影响吲哚丙酮酸、吲哚乙酰胺和吲哚乙腈途径参与吲哚-3-乙酸合成的代谢机理，为吲哚-3-乙酸合成途径解析和代谢工程育种构建吲哚-3-乙酸高产菌株奠定了基础。
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| 关 键 词： | 吲哚-3-乙酸 解淀粉芽胞杆菌 YsnE 代谢途径 |
| 收稿时间： | 2022/7/2 0:00:00 |
Metabolic pathways of ysnE involved in indole-3-acetic acid synthesis in Bacillus amyloliquefaciens HZ-12 |
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| ZHOU Huijuan,SHE Menglin,LI Ziyan,GAO Lin,GAO Jiaming,CAI Dongbo,YANG Zhifan,CHEN Shouwen,MA Xin.Metabolic pathways of ysnE involved in indole-3-acetic acid synthesis in Bacillus amyloliquefaciens HZ-12[J].Acta Microbiologica Sinica,2023,63(2):845-854. |
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| Authors: | ZHOU Huijuan SHE Menglin LI Ziyan GAO Lin GAO Jiaming CAI Dongbo YANG Zhifan CHEN Shouwen MA Xin |
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| Institution: | Environmental Microbial Technology Center of Hubei Province, State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, Hubei, China;Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong, China;Hubei Corporation of China National Tobacco Corporation, Wuhan 430000, Hubei, China |
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| Abstract: | Objective] Indole-3-acetic acid is crucial for plant growth and development and physiological activities. Indole-3-acetic acid N-acetyltransferase YsnE plays an important part in indole-3-acetic acid synthesis. This study aims to analyze the metabolic pathways of YsnE involved in indole-3-acetic acid synthesis of Bacillus amyloliquefaciens. Methods] Through deletion and overexpression of ysnE, the role of ysnE in indole-3-acetic acid synthesis was elucidated. Combined with the addition of indole-3-acetic acid synthesis intermediates (indole pyruvic acid (IPA), indole-3-acetamide (IAM), tryptamine (TAM), indole-3-acetonitrile (IAN)) and in vitro enzymatic conversion experiments, the metabolic pathways of ysnE involved in indole-3-acetic acid synthesis were clarified. Results] YsnE played an important role in indole-3-acetic acid synthesis in B. amyloliquefaciens HZ-12. The amount of IPA, IAM, and IAN consumed in ysnE deletion strain was significantly reduced, and YsnE functioned as indolepyruvate decarboxylase YclB, and indoleacetamide hydrolase/nitrilase/nitrile hydratase YhcX, and affected indole-3-acetic acid synthesis by participating in IPA, IAM, and IAN pathways. Conclusion] YsnE influences the IPA, IAM, and IAN pathways to involve in indole-3-acetic acid synthesis, which lays a foundation for analysis of indole-3-acetic acid synthesis pathway and breeding strains with high yield of indole-3-acetic acid by metabolic engineering. |
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| Keywords: | indole-3-acetic acid Bacillus amyloliquefaciens YsnE metabolic pathway |
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