Monitoring in vivo metabolic flux with a designed whole-cell metabolite biosensor of shikimic acid
Monitoring in vivo metabolic flux with a designed whole-cell metabolite biosensor of shikimic acid
复制标题
用设计的莽草酸全细胞代谢生物传感器监测体内代谢通量
DOI：
10.1016/j.bios.2017.07.022
复制
发表时间：
2017
影响因子：
12.6
通讯作者：
Tao Yong
中科院分区：
文献类型：
--
作者：
Li Heng;Liang Chaoning;Chen Wei;Jin Jian Ming;Tang Shuang Yan;Tao Yong
了解细胞内代谢物水平对于了解代谢通量分布非常重要。关键代谢物的全细胞生物传感器是监测重要代谢途径中碳流的理想选择，从而指导微生物改良的代谢工程。然而，缺乏感兴趣的代谢物的生物传感器限制了它们的应用。在这项研究中，一个基因编码的全细胞生物传感器特异性地响应莽草酸已被开发通过筛选尿酸响应调节蛋白的结合口袋的位点饱和诱变库。该生物传感器已成功地应用于分析和工程莽草酸途径的代谢通量，通过全基因组范围内筛选的基因靶点的途径通量的关键，并通过提高途径的关键酶，AroG的比活性。这项工作证明了借助针对关键代谢物设计的全细胞生物传感器监测代谢通量的可行性。
Knowledge of intracellular metabolite levels is important for the understanding of metabolic flux distributions. Whole-cell biosensors of key metabolites are ideal for the monitoring of carbon flow in important metabolic pathways, thus guiding metabolic engineering for microbial improvement. However, lack of biosensors for metabolites of interests has limited their applications. In this study, a genetically encoded whole-cell biosensor specifically responding to shikimic acid has been developed by screening a site-saturation mutagenesis library of the binding pocket of a uric acid-responsive regulatory protein. This biosensor has been successfully applied in analyzing and engineering metabolic flux in the shikimic acid pathway, through genome-wide screening of gene targets critical for the pathway flux, and by improving the specific activity of pathway key enzyme, AroG. This work demonstrates the feasibility of monitoring metabolic flux with the aid of whole-cell biosensors designed for key metabolites.