Improved 2,3-Butanediol Production Rate of Metabolically Engineered <i>Saccharomyces cerevisiae</i> by Deletion of <i>RIM15</i> and Activation of Pyruvate Consumption Pathway
<i>Saccharomyces cerevisiae</i> is a promising host for the bioproduction of higher alcohols, such as 2,3-butanediol (2,3-BDO). Metabolically engineered <i>S. cerevisiae</i> strains that produce 2,3-BDO via glycolysis have been constructed. However, the specific 2,3-BDO produ...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-11-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/22/16378 |
| Summary: | <i>Saccharomyces cerevisiae</i> is a promising host for the bioproduction of higher alcohols, such as 2,3-butanediol (2,3-BDO). Metabolically engineered <i>S. cerevisiae</i> strains that produce 2,3-BDO via glycolysis have been constructed. However, the specific 2,3-BDO production rates of engineered strains must be improved. To identify approaches to improving the 2,3-BDO production rate, we investigated the factors contributing to higher ethanol production rates in certain industrial strains of <i>S. cerevisiae</i> compared to laboratory strains. Sequence analysis of 11 industrial strains revealed the accumulation of many nonsynonymous substitutions in <i>RIM15</i>, a negative regulator of high fermentation capability. Comparative metabolome analysis suggested a positive correlation between the rate of ethanol production and the activity of the pyruvate-consuming pathway. Based on these findings, <i>RIM15</i> was deleted, and the pyruvate-consuming pathway was activated in YHI030, a metabolically engineered <i>S. cerevisiae</i> strain that produces 2,3-BDO. The titer, specific production rate, and yield of 2,3-BDO in the test tube-scale culture using the YMS106 strain reached 66.4 ± 4.4 mM, 1.17 ± 0.017 mmol (g dry cell weight h)<sup>−1</sup>, and 0.70 ± 0.03 mol (mol glucose consumed)<sup>−1</sup>. These values were 2.14-, 2.92-, and 1.81-fold higher than those of the vector control, respectively. These results suggest that bioalcohol production via glycolysis can be enhanced in a metabolically engineered <i>S. cerevisiae</i> strain by deleting <i>RIM15</i> and activating the pyruvate-consuming pathway. |
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| ISSN: | 1661-6596 1422-0067 |