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<p>This study aimed to enhance ethanol production by increasing the galactose consumption rate of recombinant <i>Saccharomyces cerevisiae</i> CEN-PK2 strains, using the CRISPR-Cas9 system from the hydrolysate of the red macroalgae <i>Eucheuma denticulatum</i>. A concentration of 27.7&#xa0;g/L of monosaccharides was obtained by thermal acid hydrolysis with 10% (w/v) biomass and 300&#xa0;mM nitric acid at 121 ℃ for 90&#xa0;min. More monosaccharide (42.0&#xa0;g/L) was obtained from subsequent enzymatic saccharification using 20 U/mL of commercial enzyme for 72&#xa0;h. The ethanol fermentation and galactose uptake rate were compared with those of the wild-type strain of <i>S. cerevisiae</i> CEN-PK2, the <i>COX9</i>- and/or <i>MIG1</i>-gene-deleted strains, and the galactose-adapted strain. A similar glucose consumption rate was observed for all tested strains. However, the <i>ΔMIG1</i> strain exhibited an enhanced galactose consumption and ethanol yield compared to the <i>ΔCOX9</i> strain and the control. Furthermore, the strain with a double deletion of <i>COX9</i> and <i>MIG1</i> (<i>ΔCOX9ΔMIG1</i>) exhibited an enhanced galactose uptake and ethanol production (31.5&#xa0;g/L, 0.46&#xa0;g/g yield, and 0.33&#xa0;g/L/h productivity). An increase in the transcription levels of the GAL regulatory genes was also observed in the <i>ΔCOX9ΔMIG1</i> strain. This research could significantly impact future energy development and climate change mitigation by enhancing bioenergy efficiency and expanding its industrial applications.</p>

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Enhancing galactose utilization and ethanol production in Saccharomyces cerevisiae via MIG1 and COX9 gene deletion using CRISPR-Cas9 with Eucheuma denticulatum hydrolysate

  • 期刊论文

  • 发表日期：

    2025年1月31日

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  • Jieun Kim,
  • In Yung Sunwoo,
  • Gwi-Taek Jeong

摘要

This study aimed to enhance ethanol production by increasing the galactose consumption rate of recombinant Saccharomyces cerevisiae CEN-PK2 strains, using the CRISPR-Cas9 system from the hydrolysate of the red macroalgae Eucheuma denticulatum. A concentration of 27.7 g/L of monosaccharides was obtained by thermal acid hydrolysis with 10% (w/v) biomass and 300 mM nitric acid at 121 ℃ for 90 min. More monosaccharide (42.0 g/L) was obtained from subsequent enzymatic saccharification using 20 U/mL of commercial enzyme for 72 h. The ethanol fermentation and galactose uptake rate were compared with those of the wild-type strain of S. cerevisiae CEN-PK2, the COX9- and/or MIG1-gene-deleted strains, and the galactose-adapted strain. A similar glucose consumption rate was observed for all tested strains. However, the ΔMIG1 strain exhibited an enhanced galactose consumption and ethanol yield compared to the ΔCOX9 strain and the control. Furthermore, the strain with a double deletion of COX9 and MIG1 (ΔCOX9ΔMIG1) exhibited an enhanced galactose uptake and ethanol production (31.5 g/L, 0.46 g/g yield, and 0.33 g/L/h productivity). An increase in the transcription levels of the GAL regulatory genes was also observed in the ΔCOX9ΔMIG1 strain. This research could significantly impact future energy development and climate change mitigation by enhancing bioenergy efficiency and expanding its industrial applications.