Mamta Gupta, Matthew Wong, Kamran Jawed, Kamil Gedeon, Hannah Barrett, Marcelo Bassalo, Clifford Morrison, Danish Eqbal, Syed Shams Yazdani, Ryan T Gill, Jiaqi Huang, Marc Douaisi, Jonathan Dordick, Georges Belfort, Mattheos A G Koffas
The production of the biofuel, isobutanol, in E. coli faces limitations due to alcohol toxicity, product inhibition, product recovery, and long-term industrial feasibility. Here we demonstrate an approach of combining both in vivo with in vitro metabolic engineering to produce isobutanol. The in vivo production of α-ketoisovalerate (KIV) was conducted through CRISPR mediated integration of the KIV pathway in bicistronic design (BCD) in E. coli and inhibition of competitive valine pathway using CRISPRi technology...
December 2022: Metabolic Engineering Communications