Engineering central carbon metabolism with phosphoketolase pathways in Saccharomyces cerevisiae
Doktorsavhandling, 2021
Secondly, we expanded the linear PK pathway to a novel configuration of the cyclic non-oxidative glycolysis (NOG) that can recycle all the carbon from glucose into acetyl-CoA, thus potentially increasing product yields even further. We showed through kinetic modeling that the new configuration resolves potential bottlenecks in the previous configuration. We verified both in vitro and in vivo functionality of the cycle in S. cerevisiae. Furthermore, we demonstrated increased titers of an acetyl-CoA-derived product in the glucose phase compared to the linear PK pathway, indicating increased precursor supply from the cycle. Finally, we further characterized the S. cerevisiae strain with the cycle, using omics. Most notably, the cycle strain yielded respiro-fermentative growth in chemostat cultures with acetate as the main overflow metabolite. This points to a metabolic imbalance and extensive AcP degradation to acetate, which needs to be resolved before the cycle can be efficiently utilized. This thesis highlights the status of this novel NOG configuration and will aid in the further development of cell factories with high-yield production of acetyl-CoA-derived products.
precursor supply
transcriptomics
sustainability
non-oxidative glycolysis
Carbon-conservation
metabolic engineering
platform strain
GATHCYC
proteomics
Författare
John Hellgren
Chalmers, Biologi och bioteknik, Systembiologi
Heterologous phosphoketolase expression redirects flux towards acetate, perturbs sugar phosphate pools and increases respiratory demand in Saccharomyces cerevisiae
Microbial Cell Factories,;Vol. 18(2019)
Artikel i vetenskaplig tidskrift
Effects of overexpression of STB5 in Saccharomyces cerevisiae on fatty acid biosynthesis, physiology and transcriptome
FEMS Yeast Research,;Vol. 19(2019)
Artikel i vetenskaplig tidskrift
Promiscuous phosphoketolase and metabolic rewiring enables novel non-oxidative glycolysis in yeast for high-yield production of acetyl-CoA derived products
Metabolic Engineering,;Vol. 62(2020)p. 150-160
Artikel i vetenskaplig tidskrift
Hellgren, J., Qi, Q., Nielsen, J. & Siewers, V. Proteome and transcriptome analysis of a yeast strain expressing a cyclic phosphoketolase pathway for improved acetyl-CoA supply
In this thesis, I have engineered S. cerevisiae to convert sugars more efficiently, with the end goal of making cell factories a competitive alternative to fossil-based sources of chemicals and fuels. This was achieved by enabling yeast to use reaction pathways that do not involve loss of carbon as CO2. I implemented a carbon-conserving pathway into yeast and investigated how this pathway affected the physiology of the yeast cell. To conserve more carbon, I expanded this linear carbon-conserving pathway to an artificial cyclic pathway that does not exist in nature. This new cycle shows great potential in increasing product yields, but one of its current limitations involves by-product formation, which needs to be addressed first. In conclusion, this work sets the foundation for the use of a novel carbon-conserving cycle and will aid in the future development of efficient cell factories.
Ämneskategorier
Biokemi och molekylärbiologi
Bioinformatik och systembiologi
Styrkeområden
Energi
ISBN
978-91-7905-580-6
Doktorsavhandlingar vid Chalmers tekniska högskola. Ny serie: 5047
Utgivare
Chalmers
Konferensrummet 10’an, Forskarhus 1, Kemigården 4, Göteborg
Opponent: Jack Pronk, Delft Technical University, Netherlands