Engineering the C3N Pathway as a Short Detour for De Novo NAD+ Biosynthesis in Saccharomyces cerevisiae
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains, Plasmids, and Growth Conditions
2.2. General DNA Manipulations and Sequence Analyses
2.3. Construction of S. cerevisiae BY01
2.4. Expression of phzD, phzE, and dhbX Individually in S. cerevisiae
2.5. Construction of S. cerevisiae BY00-Con, BY00-C3N, BY01-Con, and BY01-C3N
2.6. Growth of S. cerevisiae Strains on YMMN Plates
2.7. Measurement of Intracellular NAD(H) Concentrations
2.8. Spectroscopic Analysis
2.9. Sequence Data
3. Results
3.1. Functional Expression of the C3N Pathway Genes in S. cerevisiae
3.2. Construction of the C3N Pathway in S. cerevisiae
3.3. Expanding the NAD(H) Pool of S. cerevisiae by the C3N Pathway
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strains | NAD(H) [mM] | NAD+ [mM] |
---|---|---|
BY00 | 3.30 ± 0.45 | 1.90 ± 0.24 |
BY00-Con | 2.92 ± 0.20 | 1.40 ± 0.19 |
BY01-C3N | 3.13 ± 0.06 | 1.65 ± 0.09 |
BY00-C3N | 4.59 ± 0.37 | 2.30 ± 0.25 |
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Li, X.; Tang, Y.; Ding, Y.; Li, P.; Chen, Y. Engineering the C3N Pathway as a Short Detour for De Novo NAD+ Biosynthesis in Saccharomyces cerevisiae. Fermentation 2023, 9, 886. https://doi.org/10.3390/fermentation9100886
Li X, Tang Y, Ding Y, Li P, Chen Y. Engineering the C3N Pathway as a Short Detour for De Novo NAD+ Biosynthesis in Saccharomyces cerevisiae. Fermentation. 2023; 9(10):886. https://doi.org/10.3390/fermentation9100886
Chicago/Turabian StyleLi, Xinli, Yue Tang, Yong Ding, Pengwei Li, and Yihua Chen. 2023. "Engineering the C3N Pathway as a Short Detour for De Novo NAD+ Biosynthesis in Saccharomyces cerevisiae" Fermentation 9, no. 10: 886. https://doi.org/10.3390/fermentation9100886