强化鼠李糖合成途径基因调控丁烯基多杀菌素合成的研究  

Research on gene regulation of the synthesis of butenyl-spinosyn by enhancing rhamnose synthetic pathways

作　　者：李新颖 王靖楠 郭超 庞建 李春 王超[1] Li Xinying;Wang Jingnan;Guo Chao;Pang Jian;Li Chun;Wang Chao(Academy of National Food and Strategic Reserves Administration,Beijing 100037;Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering,Ministry of Industry and Information Technology,Institute of Biochemical Engineering,School of Chemistry and Chemical Engineering,Beijing Institute of Technology,Beijing 100081;Institute of Biochemistry,Department of chemical engineering,Tsinghua University,Beijing 100084)

机构地区：[1]国家粮食和物资储备局科学研究院粮油加工研究所,北京100037 [2]北京理工大学化学与化工学院,生物化工研究所,医药分子科学与制剂工程工信部重点实验室,北京100081 [3]清华大学化学工程系生物化工研究所,北京100084

出　　处：《中国抗生素杂志》2024年第12期1384-1391,共8页Chinese Journal of Antibiotics

基　　金：“十四五”重点研发计划项目(No.2023YFA0914700);国家自然科学基金(No.22108153);S-腺苷甲硫氨酸合成酶对丁烯基多杀菌素合成的影响(No.JY2311)。

摘　　要：目的探究鼠李糖合成途径基因强化对须糖多孢菌生长及丁烯基多杀菌素合成的影响及调控机制。方法在Saccharopolyspora pogona ASAGF58中过表达鼠李糖合成途径基因,并且在发酵培养基中添加不同浓度鼠李糖探究其对须糖多孢菌生长和次级代谢的影响。利用qRT-PCR对23个合成途径基因以及菌株生长相关基因的转录水平进行比较,挖掘影响产量的关键基因。结果通过过表达鼠李糖合成途径基因,得到3株高产菌株Rha1、Rha2和Rha3,丁烯基多杀菌素产量分别提高1.82倍、2.12倍和2.54倍。结论优化鼠李糖供应能促进丁烯基多杀菌素的合成,其中葡萄糖核苷转移酶gtt和鼠李糖基转移酶busG可能是影响丁烯基多杀菌素合成的关键基因。Objective This study explored the influence and mechanism of overexpression of rhamnose synthesis genes on the growth of Saccharopolyspora pogona and the synthesis of butenyl-spinosyn.Methods The influence of the rhamnose synthesis genes on growth and secondary metabolism was explored by overexpressing the rhamnose synthesis genes or culturing with various concentrations of rhamnose in S.pogona ASAGF58,A qRT-PCR analysis of 23 structure genes and the genes associated with cell growth was performed to identify the key genes that played an important role in yield.Results By overexpressing genes involved in the rhamnose synthesis pathway,three high-yielding strains Rha1,Rha2,and Rha3 were obtained,resulting in a 1.82 fold,2.12 fold,and 2.54 fold increase in butenyl-spinosy production,respectively.Conclusion The titer of butenyl-spinosyn could be improved by optimizing the supply of rhamnose.And gtt and rhamnosyltransferases busG were inferred to be the key genes affecting the production.

关 键 词：须糖多孢菌 丁烯基多杀菌素 鼠李糖基合成基因 工程菌构建 高产机制 

分 类 号：R978.1[医药卫生—药品]