Dabirian Yasaman, Gonçalves Teixeira Paulo, Nielsen Jens, Siewers Verena, David Florian
Department of Biology and Biological Engineering , Chalmers University of Technology , 41296 Gothenburg , Sweden.
Novo Nordisk Foundation Center for Biosustainability , Chalmers University of Technology , 41296 Gothenburg , Sweden.
ACS Synth Biol. 2019 Aug 16;8(8):1788-1800. doi: 10.1021/acssynbio.9b00118. Epub 2019 Jul 30.
Fatty acid-derived compounds have a range of industrial applications, from chemical building blocks to biofuels. Due to the highly dynamic nature of fatty acid metabolism, it is difficult to identify genes modulating fatty acyl-CoA levels using a rational approach. Metabolite biosensors can be used to screen genes from large-scale libraries in a high throughput manner. Here, a fatty acyl-CoA sensor based on the transcription factor FadR from was established in and combined with a gene overexpression library to screen for genes increasing the fatty acyl-CoA pool. Fluorescence-activated cell sorting, followed by data analysis, identified genes enhancing acyl-CoA levels. From these, overexpression of , , and resulted in about 80% increased fatty alcohol levels. Changes in fatty acid saturation and chain length distribution could also be observed. These results indicate that the use of this acyl-CoA biosensor combined with a gene overexpression library allows for identification of gene targets improving production of fatty acids and derived products.
脂肪酸衍生化合物具有一系列工业应用，从化学构建块到生物燃料。由于脂肪酸代谢具有高度动态的性质，使用合理的方法难以鉴定调节脂肪酰辅酶A水平的基因。代谢物生物传感器可用于以高通量方式从大规模文库中筛选基因。在此，基于来自[具体来源未提及]的转录因子FadR的脂肪酰辅酶A传感器在[具体生物未提及]中建立，并与基因过表达文库结合以筛选增加脂肪酰辅酶A库的基因。通过荧光激活细胞分选，随后进行数据分析，鉴定出增强酰基辅酶A水平的基因。从中，[具体基因未提及]、[具体基因未提及]和[具体基因未提及]的过表达导致脂肪醇水平增加约80%。还可观察到脂肪酸饱和度和链长分布的变化。这些结果表明，使用这种酰基辅酶A生物传感器与基因过表达文库相结合，能够鉴定出改善脂肪酸及其衍生产品产量的基因靶点。
