The coexistence of potassium and calcium ions has been identified as a major factor limiting high-yield ethanol fermentation by Saccharomyces cerevisiae in high-concentration sugarcane molasses. To identify key genes conferring tolerance to this stress, we employed an integrated strategy combining multi-omics analysis, CRISPR-mediated gene activation/repression, and targeted overexpression. This approach pinpointed four critical genes: PIR3, SPI1, AQR1, and GUT2. Functional analysis showed that while AQR1 and GUT2 enhance ethanol biosynthesis primarily by redirecting metabolic flux, PIR3 and SPI1 are crucial for maintaining cellular integrity and viability under stress. Overexpression of PIR3 and SPI1 in a wild-type strain increased ethanol production by 24.6%, achieving a final titer of 113.3 g/L in a 5-L fermenter, a performance comparable to robust industrial strains. Furthermore, this engineering strategy boosted the synthesis of other valuable compounds, exemplified by a 12.5% increase in cinnamic acid production. Our work thus identifies precise genetic targets for engineering stress-tolerant yeast and establishes a foundation for efficient bioconversion of high-concentration sugarcane molasses.
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| Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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| Site | Modification | Modifier | Source | Reference |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.
| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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| Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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| Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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