Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production
Abstract Background The thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in t...
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| Format: | Article |
| Language: | English |
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BMC
2023-04-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
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| Online Access: | https://doi.org/10.1186/s13068-023-02312-4 |
| _version_ | 1797845997503643648 |
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| author | Peizhou Yang Wenjing Wu Jianchao Chen Suwei Jiang Zhi Zheng Yanhong Deng Jiuling Lu Hu Wang Yong Zhou Yuyou Geng Kanglin Wang |
| author_facet | Peizhou Yang Wenjing Wu Jianchao Chen Suwei Jiang Zhi Zheng Yanhong Deng Jiuling Lu Hu Wang Yong Zhou Yuyou Geng Kanglin Wang |
| author_sort | Peizhou Yang |
| collection | DOAJ |
| description | Abstract Background The thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in the cell membrane of S. cerevisiae affects the thermotolerant capability. Results In this study, S. cerevisiae ERG5, ERG4, and ERG3 were knocked out using the CRISPR–Cas9 approach to impact the gene expression involved in ergosterol synthesis. The highest thermotolerant strain was S. cerevisiae ERG5ΔERG4ΔERG3Δ, which produced 22.1 g/L ethanol at 37 °C using the initial glucose concentration of 50 g/L with an increase by 9.4% compared with the wild type (20.2 g/L). The ethanol concentration of 9.4 g/L was produced at 42 ℃, which was 2.85-fold of the wild-type strain (3.3 g/L). The molecular mechanism of engineered S. cerevisiae at the RNA level was analyzed using the transcriptomics method. The simultaneous deletion of S. cerevisiae ERG5, ERG4, and ERG3 caused 278 up-regulated genes and 1892 down-regulated genes in comparison with the wild-type strain. KEGG pathway analysis indicated that the up-regulated genes relevant to ergosterol metabolism were ERG1, ERG11, and ERG5, while the down-regulated genes were ERG9 and ERG26. S. cerevisiae ERG5ΔERG4ΔERG3Δ produced 41.6 g/L of ethanol at 37 °C with 107.7 g/L of corn liquefied glucose as carbon source. Conclusion Simultaneous deletion of ERG5, ERG4, and ERG3 resulted in the thermotolerance improvement of S. cerevisiae ERG5ΔERG4ΔERG3Δ with cell viability improvement by 1.19-fold at 42 °C via modification of steroid metabolic pathway. S. cerevisiae ERG5ΔERG4ΔERG3Δ could effectively produce ethanol at 37 °C using corn liquefied glucose as carbon source. Therefore, S. cerevisiae ERG5ΔERG4ΔERG3Δ had potential in ethanol production at a large scale under supra-optimal temperature. |
| first_indexed | 2024-04-09T17:49:00Z |
| format | Article |
| id | doaj.art-2f922dd5649c4857a89021a2e8613e1c |
| institution | Directory Open Access Journal |
| issn | 2731-3654 |
| language | English |
| last_indexed | 2024-04-09T17:49:00Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj.art-2f922dd5649c4857a89021a2e8613e1c2023-04-16T11:09:13ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542023-04-0116111210.1186/s13068-023-02312-4Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol productionPeizhou Yang0Wenjing Wu1Jianchao Chen2Suwei Jiang3Zhi Zheng4Yanhong Deng5Jiuling Lu6Hu Wang7Yong Zhou8Yuyou Geng9Kanglin Wang10School of Food and Biological Engineering, Anhui Key Laboratory of Intensive Processing of Agricultural Products, Hefei University of TechnologySchool of Food and Biological Engineering, Anhui Key Laboratory of Intensive Processing of Agricultural Products, Hefei University of TechnologySchool of Food and Biological Engineering, Anhui Key Laboratory of Intensive Processing of Agricultural Products, Hefei University of TechnologyDepartment of Biological, Food and Environment Engineering, Hefei UniversitySchool of Food and Biological Engineering, Anhui Key Laboratory of Intensive Processing of Agricultural Products, Hefei University of TechnologySuzhou Cofco Biochemical Co., Ltd.Suzhou Cofco Biochemical Co., Ltd.Suzhou Cofco Biochemical Co., Ltd.Suzhou Cofco Biochemical Co., Ltd.Suzhou Cofco Biochemical Co., Ltd.Hefei Knature Bio-Pharm Co., Ltd.Abstract Background The thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in the cell membrane of S. cerevisiae affects the thermotolerant capability. Results In this study, S. cerevisiae ERG5, ERG4, and ERG3 were knocked out using the CRISPR–Cas9 approach to impact the gene expression involved in ergosterol synthesis. The highest thermotolerant strain was S. cerevisiae ERG5ΔERG4ΔERG3Δ, which produced 22.1 g/L ethanol at 37 °C using the initial glucose concentration of 50 g/L with an increase by 9.4% compared with the wild type (20.2 g/L). The ethanol concentration of 9.4 g/L was produced at 42 ℃, which was 2.85-fold of the wild-type strain (3.3 g/L). The molecular mechanism of engineered S. cerevisiae at the RNA level was analyzed using the transcriptomics method. The simultaneous deletion of S. cerevisiae ERG5, ERG4, and ERG3 caused 278 up-regulated genes and 1892 down-regulated genes in comparison with the wild-type strain. KEGG pathway analysis indicated that the up-regulated genes relevant to ergosterol metabolism were ERG1, ERG11, and ERG5, while the down-regulated genes were ERG9 and ERG26. S. cerevisiae ERG5ΔERG4ΔERG3Δ produced 41.6 g/L of ethanol at 37 °C with 107.7 g/L of corn liquefied glucose as carbon source. Conclusion Simultaneous deletion of ERG5, ERG4, and ERG3 resulted in the thermotolerance improvement of S. cerevisiae ERG5ΔERG4ΔERG3Δ with cell viability improvement by 1.19-fold at 42 °C via modification of steroid metabolic pathway. S. cerevisiae ERG5ΔERG4ΔERG3Δ could effectively produce ethanol at 37 °C using corn liquefied glucose as carbon source. Therefore, S. cerevisiae ERG5ΔERG4ΔERG3Δ had potential in ethanol production at a large scale under supra-optimal temperature.https://doi.org/10.1186/s13068-023-02312-4Thermotolerant improvementSaccharomyces cerevisiaeCRISPR–Cas9 approachTranscriptomicsEthanol productionErgosterol |
| spellingShingle | Peizhou Yang Wenjing Wu Jianchao Chen Suwei Jiang Zhi Zheng Yanhong Deng Jiuling Lu Hu Wang Yong Zhou Yuyou Geng Kanglin Wang Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production Biotechnology for Biofuels and Bioproducts Thermotolerant improvement Saccharomyces cerevisiae CRISPR–Cas9 approach Transcriptomics Ethanol production Ergosterol |
| title | Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production |
| title_full | Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production |
| title_fullStr | Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production |
| title_full_unstemmed | Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production |
| title_short | Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production |
| title_sort | thermotolerance improvement of engineered saccharomyces cerevisiae erg5 delta erg4 delta erg3 delta molecular mechanism and its application in corn ethanol production |
| topic | Thermotolerant improvement Saccharomyces cerevisiae CRISPR–Cas9 approach Transcriptomics Ethanol production Ergosterol |
| url | https://doi.org/10.1186/s13068-023-02312-4 |
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