Model-Assisted Fine-Tuning of Central Carbon Metabolism in Yeast through dCas9-Based Regulation
Artikel i vetenskaplig tidskrift, 2019

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

biosensor

CRISPR

flux balance analysis

synthetic biology

Författare

Raphael Ferreira

Chalmers, Biologi och bioteknik, Systembiologi

Christos Skrekas

Chalmers, Biologi och bioteknik, Systembiologi

Alex Hedin

Chalmers, Biologi och bioteknik, Systembiologi

Benjamín José Sánchez

Chalmers, Biologi och bioteknik, Systembiologi

Verena Siewers

Chalmers, Biologi och bioteknik, Systembiologi

Jens B Nielsen

Novo Nordisk Foundation Center for Biosustainability

Danmarks Tekniske Universitet (DTU)

Chalmers, Biologi och bioteknik, Systembiologi

Florian David

Chalmers, Biologi och bioteknik, Systembiologi

ACS Synthetic Biology

2161-5063 (eISSN)

Vol. 8 11 2457-2463

Ämneskategorier

Biokemi och molekylärbiologi

Utvecklingsbiologi

Bioinformatik och systembiologi

DOI

10.1021/acssynbio.9b00258

PubMed

31577419

Mer information

Senast uppdaterat

2019-12-02