Design and Characterization of Rapid Optogenetic Circuits for Dynamic Control in Yeast Metabolic Engineering,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Design and Characterization of Rapid Optogenetic Circuits for Dynamic Control in Yeast Metabolic Engineering
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-11-24 , DOI: 10.1021/acssynbio.0c00305
Evan M Zhao ₁ , Makoto A Lalwani ₁ , Robert J Lovelett _{1,

2} , Sergio A García-Echauri ₁ , Shannon M Hoffman ₁ , Christopher L Gonzalez ₁ , Jared E Toettcher ₃ , Ioannis G Kevrekidis _{1,

2} , José L Avalos _{1,

3,

4}

Affiliation

The use of optogenetics in metabolic engineering for light-controlled microbial chemical production raises the prospect of utilizing control and optimization techniques routinely deployed in traditional chemical manufacturing. However, such mechanisms require well-characterized, customizable tools that respond fast enough to be used as real-time inputs during fermentations. Here, we present OptoINVRT7, a new rapid optogenetic inverter circuit to control gene expression in Saccharomyces cerevisiae. The circuit induces gene expression in only 0.6 h after switching cells from light to darkness, which is at least 6 times faster than previous OptoINVRT optogenetic circuits used for chemical production. In addition, we introduce an engineered inducible GAL1 promoter (P_GAL1-S), which is stronger than any constitutive or inducible promoter commonly used in yeast. Combining OptoINVRT7 with P_GAL1-S achieves strong and light-tunable levels of gene expression with as much as 132.9 ± 22.6-fold induction in darkness. The high performance of this new optogenetic circuit in controlling metabolic enzymes boosts production of lactic acid and isobutanol by more than 50% and 15%, respectively. The strength and controllability of OptoINVRT7 and P_GAL1-S open the door to applying process control tools to engineered metabolisms to improve robustness and yields in microbial fermentations for chemical production.

中文翻译：

酵母代谢工程动态控制快速光遗传学电路的设计和表征

光遗传学在光控微生物化学生产代谢工程中的应用，提高了利用传统化学制造中常规部署的控制和优化技术的前景。然而，此类机制需要特征明确、可定制的工具，这些工具响应速度足够快，可以在发酵过程中用作实时输入。在这里，我们推出 OptoINVRT7，一种新型快速光遗传逆变器电路，用于控制酿酒酵母中的基因表达。该电路在将细胞从光照切换到黑暗后仅需 0.6 小时即可诱导基因表达，这比以前用于化学生产的 OptoINVRT 光遗传学电路至少快 6 倍。此外，我们还引入了一种工程化的诱导型GAL1启动子 (P _GAL1-S )，它比酵母中常用的任何组成型或诱导型启动子都更强。将 OptoINVRT7 与 P _GAL1-S相结合，可实现强且光可调的基因表达水平，在黑暗中诱导高达 132.9 ± 22.6 倍。这种新型光遗传学电路在控制代谢酶方面具有高性能，可将乳酸和异丁醇的产量分别提高 50% 和 15% 以上。_{OptoINVRT7 和 P GAL1-S}的强度和可控性为将过程控制工具应用于工程代谢打开了大门，以提高化学生产微生物发酵的稳健性和产量。

更新日期：2020-12-18

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11