Promiscuous phosphoketolase and metabolic rewiring enables novel non-oxidative glycolysis in yeast for high-yield production of acetyl-CoA derived products
Artikel i vetenskaplig tidskrift, 2020

Carbon-conserving pathways have the potential of increasing product yields in biotechnological processes. The aim of this project was to investigate the functionality of a novel carbon-conserving pathway that produces 3 mol of acetyl-CoA from fructose-6-phosphate without carbon loss in the yeast Saccharomyces cerevisiae. This cyclic pathway relies on a generalist phosphoketolase (Xfspk), which can convert xylulose-5-phosphate, fructose-6-phosphate and sedoheptulose-7-phosphate (S7P) to acetyl phosphate. This cycle is proposed to overcome bottlenecks from the previously reported non-oxidative glycolysis (NOG) cycle. Here, in silico simulations showed accumulation of S7P in the NOG cycle, which was resolved by blocking the non-oxidative pentose phosphate pathway and introducing Xfspk and part of the riboneogenesis pathway. To implement this, a transketolase and transaldolase deficient S. cerevisiae was generated and a cyclic pathway, the Glycolysis AlTernative High Carbon Yield Cycle (GATHCYC), was enabled through xfspk expression and sedoheptulose bisphosphatase (SHB17) overexpression. Flux through the GATHCYC was demonstrated in vitro with a phosphoketolase assay on crude cell free extracts, and in vivo by constructing a strain that was dependent on a functional pathway to survive. Finally, we showed that introducing the GATHCYC as a carbon-conserving route for 3-hydroxypropionic acid (3-HP) production resulted in a 109% increase in 3-HP titers when the glucose was exhausted compared to the phosphoketolase route only.

Mathematical modelling

Saccharomyces cerevisiae

Synthetic biology

Non-oxidative glycolysis

3-Hydroxypropionic acid

Metabolic engineering

Författare

John Hellgren

Chalmers, Biologi och bioteknik, Systembiologi

Novo Nordisk Fonden

Alexei Godina

Total S.A.

Jens B Nielsen

BioInnovation Institute

Novo Nordisk Fonden

Chalmers, Biologi och bioteknik, Systembiologi

Verena Siewers

Chalmers, Biologi och bioteknik, Systembiologi

Novo Nordisk Fonden

Metabolic Engineering

1096-7176 (ISSN) 1096-7184 (eISSN)

Vol. 62 150-160

Ämneskategorier

Annan medicinsk grundvetenskap

Mikrobiologi

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

DOI

10.1016/j.ymben.2020.09.003

PubMed

32911054

Mer information

Senast uppdaterat

2023-05-26