Enhanced production of 2,3-Butanediol by engineered Saccharomyces cerevisiae through fine-tuning of Pyruvate decarboxylase and NADH oxidase activities

Jin Woo Kim, Jungyeon Kim, Seung Oh Seo, Kyoung Heon Kim, Yong Su Jin, Jin Ho Seo

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: 2,3-Butanediol (2,3-BD) is a promising compound for various applications in chemical, cosmetic, and agricultural industries. Pyruvate decarboxylase (Pdc)-deficient Saccharomyces cerevisiae is an attractive host strain for producing 2,3-BD because a large amount of pyruvate could be shunted to 2,3-BD production instead of ethanol synthesis. However, 2,3-BD yield, productivity, and titer by engineered yeast were inferior to native bacterial producers because of the following metabolic limitations. First, the Pdc-deficient yeast showed growth defect due to a shortage of C2-compounds. Second, redox imbalance during the 2,3-BD production led to glycerol formation that lowered the yield. Results: To overcome these problems, the expression levels of Pdc from a Crabtree-negative yeast were optimized in S. cerevisiae. Specifically, Candida tropicalis PDC1 (CtPDC1) was used to minimize the production of ethanol but maximize cell growth and 2,3-BD productivity. As a result, productivity of the BD5-G1CtPDC1 strain expressing an optimal level of Pdc was 2.3 folds higher than that of the control strain in flask cultivation. Through a fed-batch fermentation, 121.8 g/L 2,3-BD was produced in 80 h. NADH oxidase from Lactococcus lactis (noxE) was additionally expressed in the engineered yeast with an optimal activity of Pdc. The fed-batch fermentation with the optimized 2-stage aeration control led to production of 154.3 g/L 2,3-BD in 78 h. The overall yield of 2,3-BD was 0.404 g 2,3-BD/g glucose which corresponds to 80.7% of theoretical yield. Conclusions: A massive metabolic shift in the engineered S. cerevisiae (BD5-G1CtPDC1-nox) expressing NADH oxidase was observed, suggesting that redox imbalance was a major bottleneck for efficient production of 2,3-BD by engineered yeast. Maximum 2,3-BD titer in this study was close to the highest among the reported microbial production studies. The results demonstrate that resolving both C2-compound limitation and redox imbalance is critical to increase 2,3-BD production in the Pdc-deficient S. cerevisiae. Our strategy to express fine-tuned PDC and noxE could be applicable not only to 2,3-BD production, but also other chemical production systems using Pdc-deficient S. cerevisiae.

Original languageEnglish
Article number265
JournalBiotechnology for Biofuels
Volume9
Issue number1
DOIs
Publication statusPublished - 2016 Jan 1

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Pyruvate Oxidase
Pyruvate Decarboxylase
Yeast
yeast
Saccharomyces cerevisiae
Tuning
productivity
fermentation
ethanol
Yeasts
Productivity
Oxidation-Reduction
production system
Fermentation
aeration
defect
glucose
Ethanol
2,3-butylene glycol
NADH oxidase

Keywords

  • 2,3-Butanediol
  • Metabolic engineering
  • Metabolomics
  • NADH oxidase
  • Pyruvate decarboxylase
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • Energy(all)
  • Management, Monitoring, Policy and Law

Cite this

Enhanced production of 2,3-Butanediol by engineered Saccharomyces cerevisiae through fine-tuning of Pyruvate decarboxylase and NADH oxidase activities. / Kim, Jin Woo; Kim, Jungyeon; Seo, Seung Oh; Kim, Kyoung Heon; Jin, Yong Su; Seo, Jin Ho.

In: Biotechnology for Biofuels, Vol. 9, No. 1, 265, 01.01.2016.

Research output: Contribution to journalArticle

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AU - Kim, Kyoung Heon

AU - Jin, Yong Su

AU - Seo, Jin Ho

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