Production of 2,3-butanediol in Saccharomyces cerevisiae by in silico aided metabolic engineering

Chiam Yu Ng, Moo Young Jung, Jinwon Lee, Min-Kyu Oh

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Background: 2,3-Butanediol is a chemical compound of increasing interest due to its wide applications. It can be synthesized via mixed acid fermentation of pathogenic bacteria such as Enterobacter aerogenes and Klebsiella oxytoca. The non-pathogenic Saccharomyces cerevisiae possesses three different 2,3-butanediol biosynthetic pathways, but produces minute amount of 2,3- butanediol. Hence, we attempted to engineer S. cerevisiae strain to enhance 2,3-butanediol production Results: We first identified gene deletion strategy by performing in silico genome-scale metabolic analysis. Based on the best in silico strategy, in which disruption of alcohol dehydrogenase (ADH) pathway is required, we then constructed gene deletion mutant strains and performed batch cultivation of the strains. Deletion of three ADH genes, ADH1, ADH3 and ADH5, increased 2,3-butanediol production by 55-fold under microaerobic condition. However, overproduction of glycerol was observed in this triple deletion strain. Additional rational design to reduce glycerol production by GPD2 deletion altered the carbon fluxes back to ethanol and significantly reduced 2,3-butanediol production. Deletion of ALD6 reduced acetate production in strains lacking major ADH isozymes, but it did not favor 2,3-butanediol production. Finally, we introduced 2,3-butanediol biosynthetic pathway from Bacillus subtilis and E. aerogenes to the engineered strain and successfully increased titer and yield. Highest 2,3-butanediol titer (2.29 g.l-1) and yield (0.113 g.g-1) were achieved by [increment]adh1 [increment]adh3 [increment]adh5 strain under anaerobic condition. Conclusions: With the aid of in silico metabolic engineering, we have successfully designed and constructed S. cerevisiae strains with improved 2,3-butanediol production.

Original languageEnglish
JournalMicrobial Cell Factories
DOIs
Publication statusAccepted/In press - 2012 May 28

Fingerprint

Metabolic engineering
Metabolic Engineering
Computer Simulation
Yeast
Saccharomyces cerevisiae
Genes
Alcohol Dehydrogenase
Alcohols
Glycerol
Enterobacter aerogenes
Biosynthetic Pathways
Gene Deletion
Chemical compounds
Bacilli
2,3-butylene glycol
Klebsiella oxytoca
Fermentation
Carbon Cycle
Bacteria
Ethanol

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Bioengineering
  • Biotechnology

Cite this

Production of 2,3-butanediol in Saccharomyces cerevisiae by in silico aided metabolic engineering. / Ng, Chiam Yu; Jung, Moo Young; Lee, Jinwon; Oh, Min-Kyu.

In: Microbial Cell Factories, 28.05.2012.

Research output: Contribution to journalArticle

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