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Subject    Enhanced production of 2,3-butanediol by engineered Saccharomyces cerevisiae through fine-tuning of pyruvate decarboxylase and NADH oxidase activities.
Jin-Woo Kim, Jin-Ho Seo. et al.
Biotechnology for Biofuels
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 and Jin-Ho Seo.

Biotechnology for Biofuels 9:265 (2016)  1-12

ABSTRACT
2,3-Butanediol (2,3-BD) is a promising compound for various applications to 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. 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. A massive metabolic shift in the engineered S. cerevisiae (BD5_G1CtPDC1_nox) expressing NADH oxidase was observed, suggesting 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.




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Subject
Name
Date
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206 2018     Elimination of biosynthetic pathways for L-valine and L-isoleucine in mitochondria enhances isobutanol production in engineered Saccharomyces cerevisiae
Kyung-Muk Lee, Sun-Ki Kim, Ye-Gi Lee, Kyung-Hye Park, Jin-Ho Seo et al.
2018/08/06  87
205 2018     Anti-melanogenic activity of schaftoside in Rhizoma Arisaematis by increasing autophagy in B16F1 cells
Pan Soo Kim, Jin-Ho Seo et al.
Biochemical and Biophysical Research Communications
2018/06/11  191
204 2018     Engineering of α−1,3-fucosyltransferases for production of 3-fucosyllactose in Escherichia coli
Jiwon Yu, Sang-Min Jung, Jin-Ho Seo and Dae-Hyuk Kweon et al.
Metabolic Engineering
2018/06/11  192
203 2017     One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis
Ling Li, Jin-ho Seo, Nam Soo Han et al.
Journal of Biotechnology
2017/10/16  790
202 2017     Enhanced production of 2,3-butanediol from xylose by combinatorial engineering of xylose metabolic pathway and cofactor regeneration in pyruvate decarboxylase-deficient Saccharomyces cerevisiae
Soo-Jung Kim,Hee-Jin Sim,Jin-Ho Seo
2017/06/07  1461
201 2017     Construction of efficient xylose-fermenting Saccharomyces cerevisiae through a synthetic isozyme system of xylose reductase from Scheffersomyces stipitis
Jung-Hyun Jo, Jin-Ho Seo et al.
Bioresource Technology
2017/05/25  1173
200 2017     Metabolic engineering of Saccharomyces cerevisiae for production of spermidine under optimal culture conditions
Sun-Ki Kim, Jung-Hyun Jo, Yong-Cheol Park, Yong-Su Jin, Jin-Ho Seo et al.
Enzyme and Microbial Technology
2017/03/22  1601
199 2017     High production of 2,3-butanediol from glycerol without 1,3-propanediol formation by Raoultella ornithinolytica B6
Taeyeon Kim, Jin-Ho Seo et al.
Applied Microbiology and Biotechnology
2017/03/20  1355
198 2016     Metabolic engineering of Saccharomyces cerevisiae for 2,3-butanediol production
Soo-Jung Kim, Jin-Ho Seo. et al.
Applied Microbiology and Biotechnology
2017/02/03  1758
197 2016     Enhanced ethanol fermentation by engineered Saccharomyces cerevisiae strains with high spermidine contents
Sun-Ki Kim, Jung-Hyun Jo, Yong-Su Jin, and Jin-Ho Seo. et al.
Bioprocess and Biosystems Engineering
2016/12/27  1586
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