Xiao Yi, Jiang Wen, Zhang Fuzhong
Department of Energy, Environmental and Chemical Engineering, ‡Division of Biological & Biomedical Sciences, §Institute of Materials Science & Engineering, Washington University in St. Louis , Saint Louis, Missouri 63130, United States.
ACS Synth Biol. 2017 Oct 20;6(10):1807-1815. doi: 10.1021/acssynbio.7b00069. Epub 2017 Jul 13.
Responding to nitrogen status is essential for all living organisms. Bacteria have evolved various complex and exquisite regulatory systems to control nitrogen metabolism. However, natural nitrogen regulatory systems, owing to their complexity, often function only in their original hosts and do not respond properly when transferred to another species. By harnessing the Lactococcus GlnRA system, we developed a genetically encoded, cross-species ammonium biosensor that displays a dynamic range up to 9-fold upon detection of ammonium ion. We demonstrated applications of this ammonium biosensor in three different species (Escherichia coli, Pseudomonas putida, and Synechocystis sp.) to detect different nitrogen sources. This ammonium sensor was further used to regulate the biosynthesis of a nitrogen-rich polymer, cyanophycin, based on ammonium concentration. Given the importance of nitrogen responses, the developed biosensor should be broadly applicable to synthetic biology and bioengineering.
对氮状态做出反应对所有生物来说都是至关重要的。细菌已经进化出各种复杂而精妙的调节系统来控制氮代谢。然而,天然的氮调节系统由于其复杂性,通常仅在其原始宿主中发挥作用,转移到另一个物种时不能正常响应。通过利用乳酸乳球菌GlnRA系统,我们开发了一种基因编码的跨物种铵生物传感器,在检测铵离子时显示出高达9倍的动态范围。我们展示了这种铵生物传感器在三种不同物种(大肠杆菌、恶臭假单胞菌和集胞藻属)中用于检测不同氮源的应用。这种铵传感器还被进一步用于根据铵浓度调节富含氮的聚合物蓝藻素的生物合成。鉴于氮响应的重要性,所开发的生物传感器应广泛应用于合成生物学和生物工程。