Design and Characterization of Rapid Optogenetic Circuits for Dynamic Control in Yeast Metabolic Engineering

Publication Year
2020

Type

Journal Article
Abstract
The use of optogenetics in metabolic engineering for light-controlled microbial chem. production raises the prospect of utilizing control and optimization techniques routinely deployed in traditional chem. 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 chem. 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%, resp. 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 chem. production
Journal
ACS Synth. Biol.
Volume
9
Pages
3254-3266
ISBN
2161-5063

CAplus AN 2020:2399656; MEDLINE PMID: 33232598 (Journal; Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.)