Cas9-based genome engineering is a powerful tool for yeast strain development, but its use in the food industry is limited due to GMO concerns. As a non-GMO alternative, adaptive laboratory evolution (ALE) was applied to enhance trehalose accumulation, a stress protectant, in Saccharomyces cerevisiae. To avoid ethanol-centric metabolism and promote storage carbohydrate production, ALE was conducted under nitrogen limitation with ethanol as the sole carbon source. The evolved strain 65EV showed a 2.3-fold increase in trehalose (11.51% vs 5.25%), resulting in enhanced cell viability (77.7% vs 5.11%) and bread loaf volume (107.2 mL vs 61.5 mL) after freeze/thaw stress. Amino acid profiling revealed distinct metabolic shifts, including elevated intracellular proline and extracellular glutamate. Whole-genome sequencing and reverse engineering identified unique mutations, TSL1 (V887A) and SSA2 (F105L), associated with trehalose regulation. These findings demonstrate potential of ALE as a non-GMO strategy for improving yeast performance in food applications.
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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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| 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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