期刊
ACS SYNTHETIC BIOLOGY
卷 9, 期 12, 页码 3254-3266出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.0c00305
关键词
-
资金
- Maeder Graduate Fellowship in Energy and the Environment
- Pew Charitable Trusts
- U.S. DOE Office of Biological and Environmental Research
- Genomic Science Program [DE-SC0019363]
- NSF CAREER Award [CBET-1751840]
- NIH [DP2EB024247]
- Schmidt Transformative Technology grant
- DARPA Lagrange Program [N66001-18-C-4031]
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 (PGAL1-S), which is stronger than any constitutive or inducible promoter commonly used in yeast. Combining OptoINVRT7 with PGAL1-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 PGAL1-S open the door to applying process control tools to engineered metabolisms to improve robustness and yields in microbial fermentations for chemical production.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据