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
李新颖 1王靖楠 1郭超 2庞建 3李春 4王超2
作者信息
- 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;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
- 2. 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
- 3. 清华大学化学工程系生物化工研究所,北京 100084
- 4. 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;清华大学化学工程系生物化工研究所,北京 100084
- 折叠
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.
Abstract
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Key words
Saccharopolyspora pogona/Butenyl-spinosyn/Rhamnose synthesis gene/Construction of engineered strains/High-yield mechanism引用本文复制引用
出版年
2024
NETL
NSTL
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