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

DC Field Value Language
dc.contributor.authorKim, Jin-Woo-
dc.contributor.authorKim, Jungyeon-
dc.contributor.authorSeo, Seung-Oh-
dc.contributor.authorKim, Kyoung Heon-
dc.contributor.authorJin, Yong-Su-
dc.contributor.authorSeo, Jin-Ho-
dc.date.accessioned2017-02-10T02:02:42Z-
dc.date.available2017-03-16T16:42:43Z-
dc.date.issued2016-12-09-
dc.identifier.citationBiotechnology for Biofuels, 9(1):265ko_KR
dc.identifier.urihttp://hdl.handle.net/10371/100674-
dc.descriptionThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made.
ko_KR
dc.description.abstractAbstract

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.
ko_KR
dc.language.isoenko_KR
dc.publisherBioMed Centralko_KR
dc.subjectPyruvate decarboxylaseko_KR
dc.subjectSaccharomyces cerevisiaeko_KR
dc.subject2, 3-Butanediolko_KR
dc.subjectNADH oxidaseko_KR
dc.subjectMetabolomicsko_KR
dc.subjectMetabolic engineeringko_KR
dc.titleEnhanced production of 2,3-butanediol by engineered Saccharomyces cerevisiae through fine-tuning of pyruvate decarboxylase and NADH oxidase activitiesko_KR
dc.typeArticleko_KR
dc.contributor.AlternativeAuthor김진우-
dc.contributor.AlternativeAuthor김정연-
dc.contributor.AlternativeAuthor서승오-
dc.contributor.AlternativeAuthor김경헌-
dc.contributor.AlternativeAuthor진용수-
dc.contributor.AlternativeAuthor서진호-
dc.identifier.doi10.1186/s13068-016-0677-9-
dc.language.rfc3066en-
dc.rights.holderThe Author(s)-
dc.date.updated2017-01-06T10:25:26Z-
Appears in Collections:
College of Agriculture and Life Sciences (농업생명과학대학)Program in Agricultural Biotechnology (협동과정-농업생물공학전공)Journal Papers (저널논문_협동과정-농업생물공학전공)
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