Model-Assisted Fine-Tuning of Central Carbon Metabolism in Yeast through dCas9-Based Regulation

• DOI: 10.1021/acssynbio.9b00258
• Publication Date: 2019-10-02T19:45:25Z
• Authors: Raphael Ferreira; Christos Skrekas; Alex Hedin; Benjamín J. Sánchez; Verena Siewers; Jens Nielsen; Florian David
• Keywords: Metabolic Engineering; Synthetic Biology; Cell Sorting

Engineering Saccharomyces cerevisiae for industrial-scale production of valuable chemicals involves extensive modulation its metabolism. Here, we identified novel gene expression fine-tuning set-ups to enhance endogenous metabolic fluxes toward increasing levels acetyl-CoA and malonyl-CoA. dCas9-based transcriptional regulation was combined together with a malonyl-CoA responsive intracellular biosensor select beneficial set-ups. The candidate genes screening were predicted using genome-scale model, gRNA library targeting total 168 selected designed. After multiple rounds fluorescence-activated cell sorting sequencing, the gRNAs that functional increased flux assessed their efficiency 3-hydroxypropionic acid (3-HP) production. 3-HP significantly improved upon involved in providing precursors, cofactor supply, as well chromatin remodeling.