Abstract: 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, the ethanol-tolerant enzyme-producing strains from humus soil wrere screened. 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. Results showed that, the mixed enzymes produced by Aspergillus niger 42 exhibited the best ethanol tolerance, and the ethanol-tolerant cellulase mixture it secreted (with an endoglucanase activity of 254±0.05 U/mL) demonstrated significant stability in 40% ethanol (P<0.001), with the relative activities of β-glucosidase and xylanase retained at 78.67%±0.25% and 55.34%±4.00%, respectively. Additionally, it does not produce ochratoxin, ensuring high safety. Response surface experiments showed that under the optimal conditions (ultrasound temperature 55 ℃, power 300 W, ethanol concentration 40%, time 30 min), using the mixed enzyme as the solution and anhydrous ethanol as the solute, the saponin yield reached 14.98%±0.08%. This represented a 101.88% increase compared to ultrasound-ethanol extraction (7.42%±0.23%), with a 30% reduction in ethanol consumption and a 59% increase in saponin yield compared to the two-step method (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.