Promoters are the most important tools to control and regulate gene expression in synthetic biology and metabolic engineering. The expression of target genes in Escherichia coli is usually controlled by inducible promoters which may cause excessive metabolic load on the host and may be uneconomical due to inducer cost. Therefore, it is important to identify more constitutive promoters that are capable of avoiding these limitations. In this study, using the monomeric red fluorescent protein (mRFP) as the reporter gene, ten promoters from the glycolytic pathway of E. coli were cloned and characterized. We found that glycolytic promoters exhibited the advantages of constitutive promoters and higher strength compared with the commonly used inducible promoter Plac. We further introduced glycolytic promoters into the mevalonate biosynthesis system. The maximum mevalonate titer produced by engineered E. coli under the control of glycolytic promoters was obviously higher than that under the control of Plac, indicating the superiority of glycolytic promoter for the metabolic engineering of E. coli. This set of glycolytic promoters significantly expands the range of engineering tools available for E. coli and can be applied in future metabolic engineering studies.

中文翻译：

---

大肠杆菌糖酵解启动子的活性评估及其在甲羟戊酸酯生物合成中的应用。

启动子是在合成生物学和代谢工程中控制和调节基因表达的最重要工具。靶基因在大肠杆菌中的表达通常由诱导型启动子控制，这些启动子可能会导致宿主体内代谢负荷过大，并且由于诱导剂成本而可能不经济。因此，重要的是确定能够避免这些限制的更多组成型启动子。在这项研究中，使用单体红色荧光蛋白（mRFP）作为报告基因，从大肠杆菌的糖酵解途径中克隆了十个启动子并进行了表征。我们发现，与常用的诱导型启动子Plac相比，糖酵解启动子显示出组成型启动子的优势和更高的强度。我们进一步将糖酵解启动子引入到甲羟戊酸酯生物合成系统中。工程改造的大肠杆菌在糖酵解启动子的控制下产生的最大甲羟戊酸效价明显高于Plac的控制下，表明糖酵解启动子对大肠杆菌的代谢工程具有优越性。这套糖酵解启动子极大地扩展了可用于大肠杆菌的工程工具的范围，可用于未来的代谢工程研究。