在Burkholderia sp. 中确定内源性强促剂 在JP2-270中使用
Jing Ke1, Jiamin Shen1, Haoran Wang1
1State Key Laboratory of Rice Biology and Breeding, China National Rice Research Institute, Hangzhou 310006, China.
Microorganisms
|September 28, 2024
概括
研究人员在Burkholderia sp.中发现了一个强大的内源性促进体. JP2-270以提高甲 (PRN) 的生产. 使用这种促进剂过度表达prnB基因增加了PRN产量1.46倍.
科学领域:
- 微生物学 微生物学
- 代谢工程是代谢工程.
- 自然产品的发现自然产品的发现
背景情况:
- 伯克霍尔迪亚物种是天然产品的重要来源,包括罗利尼特林 (PRN).
- 微生物天然产品的低产量需要代谢工程策略,如促进器工程来增强生产.
研究的目的:
- 识别和描述Burkholderia sp.中强大的内源性促进体. JP2-270用于增强基因表达.
- 通过促进剂驱动的生物合成基因过度表达来增加皮罗利尼特林 (PRN) 的产生.
主要方法:
- 用RNA测序 (RNA-seq) 来选Burkholderia sp.中高度活跃的内源性促进体. 在JP2-270.
- 使用光观测和RT-qPCR验证了促进剂活性.
- 通过顺序缩短来确定选定促进子 (P) 的最佳长度.
- 标志性促进剂被用来过度表达PRN生物合成基因 (prnA,prnB,prnC,prnD).
主要成果:
- 确定了50个具有高转录活性的内源性促进体,其中9个显示出显著的光信号.
- 一个促进子,P,被确定在三个发育时期具有高度表达.
- 对于P,确定了173bp的最小活性长度.
- 过度表达PRN生物合成基因导致基因表达增加40-80倍.
- 过度表达prnB基因导致PRN产量增加1.46倍.
结论:
- 强大的内源性促进体可以被识别并用于增强Burkholderia菌株的基因表达.
- 标志性促进物P是有效增加二次代谢物生产,特别是pyrrolnitrin.
- 这项研究为Burkholderia的代谢工程提供了宝贵的工具,用于天然产品的合成.
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