Synthetic Homoserine Lactone Sensors for Gram-Positive Bacillus subtilis Using LuxR-Type Regulators
Synthetic Homoserine Lactone Sensors for Gram-Positive Bacillus subtilis Using LuxR-Type Regulators
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DOI：
10.1021/acssynbio.3c00504
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发表时间：
2023-12-11
影响因子：
4.7
通讯作者：
Andrews，Lauren B.
中科院分区：
文献类型：
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作者：
Zeng，Min;Sarker，Biprodev;Andrews，Lauren B.
用于细菌细胞间通讯的通用生化信号可以促进不同细菌群落中的动态反应编程。然而，经典的群体感应范例是革兰氏阴性和革兰氏阳性细菌通常分别通过高丝氨酸内酯（HSL）或寡肽分子信号进行通信，以引起群体反应。在这里，我们使用变构 LuxR 型调节器（RpaR、LuxR、RhlR 和 CinR）和合成启动子创建针对革兰氏阳性枯草芽孢杆菌 168 的合成 HSL 传感器。启动子是根据不同的序列元件（-35、-16、-10和转录起始区）组合设计的。我们量化了这些组合启动子对传感器活性的影响，并确定了调节器表达如何影响其激活，实现了高达 293 倍的激活。利用实验的统计设计，我们确定了启动子区域和成对相互作用对传感器活性的显着影响，这有助于理解合成启动子设计的序列-功能关系。我们在 B 中展示了第一组已知的功能性 HSL 传感器（≥20 倍动态范围）。 subtilis 具有四种不同的 HSL 化学信号：对香豆酰基-HSL、3-氧代己酰基-HSL、正丁酰基-HSL 和正-(3-羟基十四烷酰基)-HSL。这套用于革兰氏阳性细菌的合成 HSL 传感器可以为微生物群落内可设计的种间通信铺平道路。
A universal biochemical signal for bacterial cell–cell communication could facilitate programming dynamic responses in diverse bacterial consortia. However, the classical quorum sensing paradigm is that Gram-negative and Gram-positive bacteria generally communicate via homoserine lactones (HSLs) or oligopeptide molecular signals, respectively, to elicit population responses. Here, we create synthetic HSL sensors for Gram-positiveBacillus subtilis168 using allosteric LuxR-type regulators (RpaR, LuxR, RhlR, and CinR) and synthetic promoters. Promoters were combinatorially designed from different sequence elements (−35, −16, −10, and transcriptional start regions). We quantified the effects of these combinatorial promoters on sensor activity and determined how regulator expression affects its activation, achieving up to 293-fold activation. Using the statistical design of experiments, we identified significant effects of promoter regions and pairwise interactions on sensor activity, which helped to understand the sequence–function relationships for synthetic promoter design. We present the first known set of functional HSL sensors (≥20-fold dynamic range) inB. subtilisfor four different HSL chemical signals:p-coumaroyl-HSL, 3-oxohexanoyl-HSL,n-butyryl-HSL, andn-(3-hydroxytetradecanoyl)-HSL. This set of synthetic HSL sensors for a Gram-positive bacterium can pave the way for designable interspecies communication within microbial consortia.