School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore County, Baltimore, MD, 21250, USA.
Metab Eng. 2020 Jul;60:66-76. doi: 10.1016/j.ymben.2020.03.006. Epub 2020 Mar 26.
Erythritol is an important sweetener ingredient and chemical precursor for synthesizing materials with phase transition behavior. Commercial erythritol is primarily produced by industrial fermentation. Further strain engineering necessitates the development of high throughput screening method for rapid detection and screening of mutant strain libraries. In this work, we took advantage of the erythritol-responsive transcription factor EryD, and constructed a sensor-regulator system for rapid screening and characterization of erythritol overproducers. We configured the optimal architecture of the EryD sensor-regulator construct with improved sensitivity, specificity and dynamic response range. Coupled with mutagenesis and strain screening based on biosensors, we rapidly screened and characterized a strain library containing 1152 mutants derived from combined UV and ARTP mutagenesis, in a relatively short period of time (1 week). The optimal strain produced more than 148 g/L erythritol in bench-top reactors. This work provides a reference for other metabolic engineering researchers to develop industrially-relevant strains. The reported framework enables us to rapidly improve strain performance and engineer efficient microbial cell factories for industrial applications.
赤藓糖醇是一种重要的甜味剂成分和化学前体，可用于合成具有相变行为的材料。商业赤藓糖醇主要通过工业发酵生产。进一步的菌株工程需要开发高通量筛选方法，以快速检测和筛选突变株文库。在这项工作中，我们利用赤藓糖醇响应转录因子 EryD，构建了一个传感器-调节剂系统，用于快速筛选和表征赤藓糖醇高产菌株。我们优化了 EryD 传感器-调节剂构建体的最佳结构，提高了其灵敏度、特异性和动态响应范围。结合生物传感器的诱变和菌株筛选，我们在相对较短的时间（1 周）内快速筛选和表征了包含 1152 个突变体的突变株文库，这些突变体来自于 UV 和 ARTP 联合诱变。最佳菌株在台式反应器中产生的赤藓糖醇超过 148 g/L。这项工作为其他代谢工程研究人员开发工业相关菌株提供了参考。所报道的框架使我们能够快速提高菌株性能，并设计高效的微生物细胞工厂，用于工业应用。
