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Inhibitors including sugar degradation products (e.g., 5-hydroxymethylfurfural and furfural) and phenols (e.g., 4-hydroxybenzoic acid and vanillin) from lignin degradation are inevitably formed in the pretreatment process of lignocellulose raw materials, exerting a negative impact on the fermentation efficiency. [Objective] To improve the tolerance of yeast to inhibitors in cellulose hydrolysates and ensure the efficient production of industrial biomass ethanol. [Methods] The model strain W303-1A was domesticated with the inhibitor furfural and p-hydroxybenzoic acid alone or in combination. The growth curves and ethanol fermentation performance of the domesticated strain and the original strain were compared under different inhibitor concentrations. We then conducted high-throughput genome resequencing of both the domesticated and original strains to identify the mutations in genes related to the glucose metabolism and drug resistance, thereby analyzing the variation points related to inhibitor tolerance. [Results] In the medium containing 2.0 g/L furfural, the ethanol yield of F-2 was 19.40 g/L, which was 2 times higher than that of the original strain. In the medium containing 1.6 g/L furfural and p-hydroxybenzoic acid, the highest ethanol yield of B-2 was 20.22 g/L, 7.6 times that of the original strain. Then, high-throughput genome resequencing of the original and domesticated strains revealed several mutations in the genes encoding ethanol dehydrogenase, fructose-1, 6-diphosphate aldolase, and pyruvate dehydrogenase in the glucose metabolism pathway. The mutations of YAP1 (transcriptional activator involved in oxidative stress response and REDOX homeostasis), PDR5 (pleiotropic ABC transporter tolerant to multiple chemicals), and RPN4 (zinc finger protein) genes played an important role in the inhibitor tolerance of Saccharomyces cerevisiae. [Conclusion] The findings provide more targets for further optimization and construction of model strains.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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