The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
Evolva SA, Reinach, Switzerland.
Nat Chem Biol. 2016 Nov;12(11):951-958. doi: 10.1038/nchembio.2177. Epub 2016 Sep 19.
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.
全细胞生物催化剂已被证明是一种可行的方法，可用于可持续生产大宗化学品和精细化学品。然而，为了筛选出表现最佳的生物催化剂，对细胞设计文库进行筛选通常是一项低通量的工作。出于这个原因，实时监测生产过程的生物传感器的开发引起了人们的关注。在这里，我们应用了多个参数的系统工程，在芽殖酵母酿酒酵母中搜索基于小分子结合转录激活因子的通用生物传感器设计，这些小分子结合转录激活因子来自原核超级家族的 LysR 型转录调节剂（LTTRs）。我们确定了一种设计方案，支持在存在同源小分子诱导剂的情况下，LTTR 依赖的报告基因表达激活。作为原理验证，我们将生物传感器应用于体内筛选以不同水平生产柚皮素或顺式，顺式-粘康酸的细胞，并发现报告基因的输出与产量相关。原核转录激活因子向真核底盘的移植说明了一种迄今尚未开发的生物传感器资源的潜力，该资源可用于生物技术应用。
