Expression of styAB is regulated by a two-component system during indigo biosynthesis in Pseudomonas putida
Expression of styAB is regulated by a two-component system during indigo biosynthesis in Pseudomonas putida
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恶臭假单胞菌靛蓝生物合成过程中 styAB 的表达受到双组分系统的调节
DOI：
10.1016/j.bbrc.2019.08.142
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发表时间：
2019-10-29
影响因子：
3.1
通讯作者：
Wang, Chengtao
中科院分区：
文献类型：
--
作者：
Cheng, Lei;Yue, Jianming;Wang, Chengtao
涉及StyS和StyR的双组分调控系统参与了假单胞菌中靛蓝合成的调控。然而，StyR基因在靛蓝合成中的功能和StyR增强styAB操纵子表达的详细机制尚不清楚。因此，在本研究中，我们构建了styR/styS基因敲除突变株。通过比较野生型和突变株之间靛蓝产量的差异，我们发现styR基因突变株没有靛蓝合成能力，而野生型菌株的产量为5.4 mg/L。因此，这些发现表明styR基因在靛蓝合成的调节中起着关键作用。通过电泳迁移率变动分析验证了StyS、StyR和styAB启动子之间的相互作用。结果表明，StyR通过与styAB启动子中的回文结合而与styAB启动子相互作用。此外，在恶臭假单胞菌B3的靛蓝生物合成过程中，StyS的激酶功能通过转磷酸化StyR来调节StyR。总之，这些发现为利用恶臭假单胞菌建立环境友好的靛蓝合成方法提供了重要见解。(C)2019爱思唯尔公司All rights reserved.
A two-component regulatory system involving StyS and StyR is involved in the regulation of indigo synthesis in Pseudomonas sp. However, the function of the styR gene in indigo synthesis and the detailed mechanisms through which StyR enhances the expression of the styAB operon are unclear. Accordingly, in this study, we constructed a styR/styS gene knockout mutant strain. By comparing the differences in indigo yields between the wild-type and mutant strains, we found that the styR gene mutant strain had no indigo synthesis ability, whereas the yield in the wild-type strain was 5.4 mg/L. Thus, these findings indicate that the styR gene plays a key role in the regulation of indigo synthesis. The interactions among StyS, StyR, and the styAB promoter were verified by electrophoresis mobility shift assays. The results showed that StyR interacts with the styAB promoter by binding to the palindrome in the styAB promoter. Moreover, the kinase function of StyS regulated StyR by transphosphorylating StyR during indigo biosynthesis in P. putida B3. Taken together, these findings provide important insights into the establishment of environmentally friendly indigo synthesis methods using P. putida. (C) 2019 Elsevier Inc. All rights reserved.