Enhanced production of 2,3-butanediol by engineered Saccharomyces cerevisiae through fine-tuning of pyruvate decarboxylase and NADH oxidase activities
DC Field | Value | Language |
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dc.contributor.author | Kim, Jin-Woo | - |
dc.contributor.author | Kim, Jungyeon | - |
dc.contributor.author | Seo, Seung-Oh | - |
dc.contributor.author | Kim, Kyoung Heon | - |
dc.contributor.author | Jin, Yong-Su | - |
dc.contributor.author | Seo, Jin-Ho | - |
dc.date.accessioned | 2021-09-03T15:51:39Z | - |
dc.date.available | 2021-09-03T15:51:39Z | - |
dc.date.created | 2021-06-16 | - |
dc.date.issued | 2016-12-09 | - |
dc.identifier.issn | 1754-6834 | - |
dc.identifier.uri | https://scholar.korea.ac.kr/handle/2021.sw.korea/86555 | - |
dc.description.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 C-2-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 C-2-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. | - |
dc.language | English | - |
dc.language.iso | en | - |
dc.publisher | BMC | - |
dc.subject | LIMITED CHEMOSTAT CULTURES | - |
dc.subject | GLUCOSE | - |
dc.subject | GROWTH | - |
dc.subject | EXPRESSION | - |
dc.subject | DEHYDROGENASE | - |
dc.subject | FERMENTATION | - |
dc.subject | METABOLISM | - |
dc.subject | DELETION | - |
dc.subject | IMPACT | - |
dc.subject | ENZYME | - |
dc.title | Enhanced production of 2,3-butanediol by engineered Saccharomyces cerevisiae through fine-tuning of pyruvate decarboxylase and NADH oxidase activities | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Kim, Kyoung Heon | - |
dc.identifier.doi | 10.1186/s13068-016-0677-9 | - |
dc.identifier.scopusid | 2-s2.0-85013396269 | - |
dc.identifier.wosid | 000390716400001 | - |
dc.identifier.bibliographicCitation | BIOTECHNOLOGY FOR BIOFUELS, v.9 | - |
dc.relation.isPartOf | BIOTECHNOLOGY FOR BIOFUELS | - |
dc.citation.title | BIOTECHNOLOGY FOR BIOFUELS | - |
dc.citation.volume | 9 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalResearchArea | Energy & Fuels | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Energy & Fuels | - |
dc.subject.keywordPlus | LIMITED CHEMOSTAT CULTURES | - |
dc.subject.keywordPlus | GLUCOSE | - |
dc.subject.keywordPlus | GROWTH | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | DEHYDROGENASE | - |
dc.subject.keywordPlus | FERMENTATION | - |
dc.subject.keywordPlus | METABOLISM | - |
dc.subject.keywordPlus | DELETION | - |
dc.subject.keywordPlus | IMPACT | - |
dc.subject.keywordPlus | ENZYME | - |
dc.subject.keywordAuthor | Pyruvate decarboxylase | - |
dc.subject.keywordAuthor | Saccharomyces cerevisiae | - |
dc.subject.keywordAuthor | 2, 3-Butanediol | - |
dc.subject.keywordAuthor | NADH oxidase | - |
dc.subject.keywordAuthor | Metabolomics | - |
dc.subject.keywordAuthor | Metabolic engineering | - |
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