LytSR和PdtaSR双组分系统对须糖多孢菌丁烯基多杀菌素生物合成的影响

The effect of LytSR and PdtaSR two-component systems on the biosynthesis of butenyl-spinosyn of Saccharopolyspora pogona

摘要:

【目的】须糖多孢菌（Saccharopolyspora pogona）基因组中存在一些双组分系统（two-component system，TCS），包括一个LytR调控因子和一个PdtaR转录抗终止调节因子，我们旨在通过基因工程技术改造分析这两个双组分系统蛋白在须糖多孢菌中的作用。【方法】本研究利用融合PCR和属间接合转移技术，构建了S. pogona-∆lytR和S. pogona-PdtaR工程菌株，研究它们对须糖多孢菌丁烯基多杀菌素生物合成的影响。【结果】HPLC检测和质谱鉴定表明，与原始菌相比，S. pogona-∆lytR中丁烯基多杀菌素产量有所提高，而S. pogona-PdtaR中产量下调，表明lytR和pdtaR对丁烯基多杀菌素生物合成有负调控作用。此外，生长曲线测定和表型分析结果表明，S. pogona-∆lytR和S. pogona-PdtaR对须糖多孢菌的生长发育和孢子形成也产生了一定的影响。【结论】本研究为进一步阐明丁烯基多杀菌素生物合成的调控机制奠定了理论基础。

关键词:须糖多孢菌;丁烯基多杀菌素;双组分系统;调控因子

Abstract:

[Objective] There are some two-component systems in the genome of Saccharopolyspora pogona, including a LytR regulatory factor and a PdtaR transcription resistance termination regulator, which were analyzed through genetic engineering technology.[Methods] In this study, we performed the fusion PCR and inter-genus conjugative transfer technology to construct S. pogona-∆lytR and S. pogona-PdtaR, and studied their effects on the biosynthesis of butenyl-spinosyn.[Results] We found that the production of butenyl-spinosyn in S. pogona-∆lytR was increased compared that in the wild type, while the butenyl-spinosyn production of S. pogona-PdtaR was dropt through high performance liquid chromatography (HPLC) experiments, indicating that lytR and pdtaR negetively regulating the biosynthesis of butenyl-spinosyn. In addition, we found that S. pogona-∆lytR and S. pogona-PdtaR also had certain impacts on the growth and sporulation.[Conclusion] This stydy laid a theoretical foundation for further elucidating the regulatory mechanism of butenyl-spinosyn biosynthesis.

Key words:Saccharopolyspora pogona;butenyl-spinosyn;two-component system;regulatory factor