PT  - JOURNAL ARTICLE
AU  - Haoqi Chen
AU  - Lianne Mulder
AU  - Hein J. Wijma
AU  - Ronja Wabeke
AU  - Jose Pedro Vila Cha Losa
AU  - Mattia Rovetta
AU  - Tijn Caspar de Leeuw
AU  - Andreas Millias-Argeitis
AU  - Matthias Heinemann
TI  - A photo-switchable yeast isocitrate dehydrogenase to control metabolic flux through the citric acid cycle
AID  - 10.1101/2021.05.25.445643
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.05.25.445643
4099  - http://biorxiv.org/content/early/2021/05/25/2021.05.25.445643.short
4100  - http://biorxiv.org/content/early/2021/05/25/2021.05.25.445643.full
AB  - For various research questions in metabolism, it is highly desirable to have means available, with which the flux through specific pathways can be perturbed dynamically, in a reversible manner, and at a timescale that is consistent with the fast turnover rates of metabolism. Optogenetics, in principle, offers such possibility. Here, we developed an initial version of a photo-switchable isocitrate dehydrogenase (IDH) aimed at controlling the metabolic flux through the citric acid cycle in budding yeast. By inserting a protein-based light switch (LOV2) into computationally identified active/regulatory-coupled sites of IDH and by using in vivo screening in Saccharomyces cerevisiae, we obtained a number of IDH enzymes whose activity can be switched by light. Subsequent in-vivo characterization and optimization resulted in an initial version of photo-switchable (PS) IDH. While further improvements of the enzyme are necessary, our study demonstrates the efficacy of the overall approach from computational design, via in vivo screening and characterization. It also represents one of the first few examples, where optogenetics were used to control the activity of a metabolic enzyme.Competing Interest StatementThe authors have declared no competing interest.