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Objective To identify high-quality yeast strains in the Xinjiang traditional milk wine fermentation system and screen specialized strains suitable for the milk wine fermentation, thereby providing a theoretical basis and strain reserves for the development of dedicated fermentation agents. Methods With alcohol production, ester production, lactose utilization ability, as well as acid (pH), glucose, and ethanol tolerance as screening criteria, the gradient dilution separation method combined with selective media was employed to isolate yeast strains. Morphological observation and molecular biological identification were conducted to determine the taxonomic status of the strains. The growth characteristics, carbon source utilization ability, and biosafety of the strains were investigated. The fermentation flavor contributions of the strains were evaluated through sensory assessment and electronic nose technology. Results Forty yeast strains were isolated and purified from Xinjiang traditional milk wine starter and traditional fermented dairy products. After multiple rounds of screening, two strains (J17 and J23) with excellent functions were obtained, tolerating pH 2.5, 350 g/L glucose, and 47.34 g/L ethanol. They were identified as Kluyveromyces marxianus and Pichia kudriavzevii,respectively. The growth curves of the two strains showed that the logarithmic phase began at the time point of 4 h and the stationary phase started at 12 h and 18 h, respectively. The co-culture test confirmed no antagonistic effects and demonstrated symbiotic relationships between the two strains. Carbon source utilization tests indicated that both strains efficiently utilized seven carbon sources, including glucose, lactose, and sucrose, demonstrating strong metabolic adaptability. Biosafety testing revealed that neither strain exhibited hemolytic activity and was sensitive to antifungal drugs such as ketoconazole, meeting the safety standards for food fermentation strains. Electronic nose analysis revealed that the 1:1 mixed strain (HJ) fermentation of milk wine substrate exhibited significantly higher response values for flavor compounds such as alkanes, sulfides, alcohols, and aldehydes/ketones than the single-strain fermentation groups and controls. Both the cumulative contribution rate of PCA and the cumulative discriminant rate of LDA reached 99.97%. Sensory assessment demonstrated that the HJ fermented milk wine scored higher than that of other strain combinations in four dimensions: appearance, aroma, taste, and style. Conclusion K. marxianus J17 and P. kudriavzevii J23 demonstrate high tolerance, broad-spectrum carbon source utilization ability, excellent biosafety, and synergistic aroma enhancement, showing the potential as specialized fermentation agents for milk wine production and providing high-quality microbial resources for the fermentation of traditional milk wine.
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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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