Development of a growth-sensitive malonyl-CoA platform strain

Abstract

The transition from a fossil reliance society towards full reliance on sustainable alternatives is a difficult challenge. The development of high-performant microbial cell factories, which efficiency convert renewable carbon sources to generate industrially relevant compounds, are to date not economically compatible with the current less sustainable alternatives. Through metabolic reprogramming, the production of compounds at industrial levels could be obtained by increasing cofactors and substrates levels. In this thesis, we developed a growth-sensitive malonyl-CoA platform strain, able to select for an enrichment of the highly relevant precursor, malonyl-CoA. Through coupling it with a CRISPR-dCas9 gRNA library we performed a biosensor assisted laboratory evolution (BALE). Three targets, previously described to significantly increase the downstream product of malonylCoA, 3-Hydroxypropionic acid, were selected after 24 hours and more targets will be characterized.