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Ethanol-tolerant Enzyme Preparation and One-step Efficient Extraction of Saponins from Xanthoceras sorbifolium Leaves
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Huan LONG1, Rui GUO1, Jiahong WANG1, 2, 3, *
Science and Technology of Food Industry | 2026, 47(8) : 273 - 284
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Science and Technology of Food Industry | 2026, 47(8): 273-284
• Processing Technology •
Ethanol-tolerant Enzyme Preparation and One-step Efficient Extraction of Saponins from Xanthoceras sorbifolium Leaves
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Huan LONG1, Rui GUO1, Jiahong WANG1, 2, 3, *
Affiliations
  • 1National Key Laboratory for the Development and Utilization of Forest Food Resources, Nanjing Forestry University, Nanjing 210037, China
  • 2Co-innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
  • 3Co-innovation Center of Efficient Procession of Forest Resources, Nanjing Forestry University, Nanjing 210037, China
Published: 2026-04-15 doi: 10.13386/j.issn1002-0306.2025050306
Outline
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To address the current issues of high consumption of organic solvent and prolonged extraction time inherent in the two-step method involving enzymes and ultrasound for efficient extraction of saponins from Xanthoceras sorbifolium leaves. A specific strain of black aspergillus fungus from humus soil was selectively isolated in this study. Based on the ethanol tolerance characteristics of the mixed enzymes obtained after fermentation, a one-step method using ultrasonication-enzyme-ethanol extraction process for saponins from Xanthoceras sorbifolium leaves was designed and compared with the ultrasonication-assisted enzymatic extraction (two-step method). The optimized conditions through single factor and response surface experiments were employed to analyze samples using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD) to characterize and analyze the structural features of the Xanthoceras sorbifolium leaf residue. As a results, the ethanol-tolerant cellulase mixed enzyme secreted by this strain, exhibiting an endoglucanase activity of 254±0.05 U/mL, which was demonstrated remarkable stability in 40% ethanol, with relative enzyme activities of β-glucosidase and xylanase retained at 78.67%±0.25% and 55.34%±4.00%, respectively. Importantly, the mixed enzyme does not contain ochratoxin, ensuring high safety. Under optimal conditions (ultrasonic temperature 55 ℃, power 300 W, ethanol concentration 40%, time 30 min) with mixed enzyme as a solution and anhydrous ethanol as a solvent, the response surface experiments revealed that saponin yield reached 14.98%±0.08%, representing a 101.89% increase compared to ultrasonic ethanol extraction (7.42%±0.23%). Furthermore, ethanol consumption was reduced by 30% relative to the two-step method, while saponin yield improved by 59% (9.42%±0.04%). UPLC-Q-TOF MS analysis confirmed that the saponin composition unchanged remained from one-step method and showing a significant increase in content compared to the two step method. Besides, SEM, FT-IR and XRD further confirmed the effectiveness of one-step method, inducing more substantial material degradation than the two-step method, more pronounced disruption of O-H, C=C and C-O-C bonds and complete degradation of amorphous regions. This one-step method combined the advantages of high saponin yield, low solvent consumption, mild operating conditions, and environmental friendliness, providing a novel solution for the green and efficient extraction of saponins from Xanthoceras sorbifolium leaves and offering valuable insight for the extraction of other natural active compounds.

strains screening  /  ethanol-tolerant cellulase  /  saponins from Xanthoceras sorbifolium Bunge leaves  /  enzymatic characteristics  /  one-step method  /  process optimization
龙欢, 郭锐, 王佳宏. 耐乙醇酶制备及一步法高效提取文冠果叶皂苷. 食品工业科技, 2026 , 47 (8) : 273 -284 . DOI: 10.13386/j.issn1002-0306.2025050306
Huan LONG, Rui GUO, Jiahong WANG. Ethanol-tolerant Enzyme Preparation and One-step Efficient Extraction of Saponins from Xanthoceras sorbifolium Leaves[J]. Science and Technology of Food Industry, 2026 , 47 (8) : 273 -284 . DOI: 10.13386/j.issn1002-0306.2025050306
Year 2026 volume 47 Issue 8
121
60
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doi: 10.13386/j.issn1002-0306.2025050306
  • Receive Date:2025-05-29
  • Online Date:2026-10-24
  • Published:2026-04-15
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  • Received:2025-05-29
Affiliations
    1National Key Laboratory for the Development and Utilization of Forest Food Resources, Nanjing Forestry University, Nanjing 210037, China
    2Co-innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
    3Co-innovation Center of Efficient Procession of Forest Resources, Nanjing Forestry University, Nanjing 210037, China
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表12种不同金属材料的力学参数
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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