Design and construction of acetyl-CoA overproducing Saccharomyces cerevisiae strains

Jiazhang Lian, Tong Si, Nikhil U. Nair, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Saccharomyces cerevisiae has increasingly been engineered as a cell factory for efficient and economic production of fuels and chemicals from renewable resources. Notably, a wide variety of industrially important products are derived from the same precursor metabolite, acetyl-CoA. However, the limited supply of acetyl-CoA in the cytosol, where biosynthesis generally happens, often leads to low titer and yield of the desired products in yeast. In the present work, combined strategies of disrupting competing pathways and introducing heterologous biosynthetic pathways were carried out to increase acetyl-CoA levels by using the CoA-dependent n-butanol production as a reporter. By inactivating ADH1 and ADH4 for ethanol formation and GPD1 and GPD2 for glycerol production, the glycolytic flux was redirected towards acetyl-CoA, resulting in 4-fold improvement in n-butanol production. Subsequent introduction of heterologous acetyl-CoA biosynthetic pathways, including pyruvate dehydrogenase (PDH), ATPdependent citrate lyase (ACL), and PDH-bypass, further increased n-butanol production. Recombinant PDHs localized in the cytosol (cytoPDHs) were found to be the most efficient, which increased n-butanol production by additional 3 fold. In total, n-butanol titer and acetyl-CoA concentration were increased more than 12 fold and 3 fold, respectively. By combining the most effective and complementary acetyl-CoA pathways, more than 100 mg/L n-butanol could be produced using high cell density fermentation, which represents the highest titer ever reported in yeast using the clostridial CoA-dependent pathway.

Original languageEnglish (US)
Title of host publicationFood, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting
PublisherAIChE
Pages750-760
Number of pages11
ISBN (Electronic)9781510812598
DOIs
StatePublished - Jan 1 2014
EventFood, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting - Atlanta, United States
Duration: Nov 16 2014Nov 21 2014

Publication series

NameFood, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting
Volume2

Other

OtherFood, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting
CountryUnited States
CityAtlanta
Period11/16/1411/21/14

Keywords

  • ATP-dependent citrate lyase
  • Acetyl-CoA
  • Acetyl-CoA synthetase
  • Metabolic engineering
  • N-Butanol
  • Pyruvate dehydrogenase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Pharmaceutical Science
  • Bioengineering

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  • Cite this

    Lian, J., Si, T., Nair, N. U., & Zhao, H. (2014). Design and construction of acetyl-CoA overproducing Saccharomyces cerevisiae strains. In Food, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting (pp. 750-760). (Food, Pharmaceutical and Bioengineering Division 2014 - Core Programming Area at the 2014 AIChE Annual Meeting; Vol. 2). AIChE. https://doi.org/10.1016/j.ymben.2014.05.010