| 초록 |
High-throughput screening is important for combinatorial metabolic engineering to efficiently investigate vast phenotypic diversity. However, current methods place constraints on monitoring of the intracellular concentration of diverse target products at single cell level. Here, we establish a high-throughput screening platform for metabolite production based on microfluidic static droplet array (SDA) and artificial riboswitch. In this system, a genetically encoded riboswitch enables the measurement of the intracellular metabolite concentration through fluorescence. Then, single cells of a library are individually entrapped in SDA to detect and collect highly productive variants. Using this approach, we analyzed L-tryptophan concentrations of Escherichia coli cells. A mutant E. coli library was screened for improved L-tryptophan production and superior strains were obtained exhibiting up to 145% productivity compared to their parental strain. |
