计算生物学
生物传感器
酿酒酵母
报告基因
生物化学
酵母
生物
高通量筛选
小分子
诱导剂
定向进化
代谢工程
合成生物学
细胞生物学
转录调控
基因
转录因子
基因表达
突变体
作者
M. Skjoedt,Tim Snoek,Kanchana Rueksomtawin Kildegaard,Dushica Arsovska,Michael Eichenberger,Tobias Justus Goedecke,Arun S. Rajkumar,Jie Zhang,Mette Kristensen,Beata Joanna Lehka,Solvej Siedler,Irina Borodina,Michael K. Jensen,Jay D. Keasling
标识
DOI:10.1038/nchembio.2177
摘要
Transplantation of the prokaryotic LysR-type transcriptional regulator into yeast combined with in vivo screening identifies yeast mutants that produce metabolic products with bacterial small molecule inducers. Whole-cell biocatalysts have proven a tractable path toward sustainable production of bulk and fine chemicals. Yet the screening of libraries of cellular designs to identify best-performing biocatalysts is most often a low-throughput endeavor. For this reason, the development of biosensors enabling real-time monitoring of production has attracted attention. Here we applied systematic engineering of multiple parameters to search for a general biosensor design in the budding yeast Saccharomyces cerevisiae based on small-molecule binding transcriptional activators from the prokaryote superfamily of LysR-type transcriptional regulators (LTTRs). We identified a design supporting LTTR-dependent activation of reporter gene expression in the presence of cognate small-molecule inducers. As proof of principle, we applied the biosensors for in vivo screening of cells producing naringenin or cis,cis-muconic acid at different levels, and found that reporter gene output correlated with production. The transplantation of prokaryotic transcriptional activators into the eukaryotic chassis illustrates the potential of a hitherto untapped biosensor resource useful for biotechnological applications.
科研通智能强力驱动
Strongly Powered by AbleSci AI