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草业学报 ›› 2026, Vol. 35 ›› Issue (7): 201-216.DOI: 10.11686/cyxb2025291

• 研究论文 • 上一篇    

苜蓿皂苷发酵和超声提取工艺条件优化研究

李武1(), 李佳静1(), 李洁冰1, 李振舟4, 郭明1,2,3, 牛岩5, 孙浩1,2,3, 李德锋1,2,3, 史莹华1,2,3, 李振田1,2,3, 刘伯帅1,2,3, 崔亚垒1,2,3, 王志昌1,2,3, 李永涛4, 朱晓艳1,2,3()   

  1. 1.河南农业大学动物科技学院,河南 郑州 450046
    2.河南省草地资源创新与利用重点实验室,河南 郑州 450046
    3.河南省牧草工程技术研究中心,河南 郑州 450046
    4.河南农业大学动物医学院,河南 郑州 450046
    5.河南省畜牧局饲草饲料站,河南 郑州 450002
  • 收稿日期:2025-07-14 修回日期:2025-10-09 出版日期:2026-07-20 发布日期:2026-05-21
  • 通讯作者: 朱晓艳
  • 作者简介:Corresponding author. E-mail: zxy_0512@163.com
    李武(1998-),男,河南信阳人,在读硕士。E-mail: 15294728201@163.com
    李佳静(1998-),女,河南新乡人,硕士。E-mail: jingl5464@163.com第一联系人:共同第一作者These authors contributed equally to this work.
  • 基金资助:
    河南省高等学校重点科研项目(26A230010);2023年度河南农业大学科技创新基金项目(2023CXZX010);2024年第五批河南省创新研发专项子课题(30603371);国家留学基金青年骨干教师出国研修项目(留金项〔2023〕54号);国家现代农业产业技术体系(CARS-34);河南省优质饲草与动物健康科技创新团队(22IRTSTHN022)

Optimization of fermentation and ultrasound-assisted extraction parameters for alfalfa saponins

Wu LI1(), Jia-jing LI1(), Jie-bing LI1, Zhen-zhou LI4, Ming GUO1,2,3, Yan NIU5, Hao SUN1,2,3, De-feng LI1,2,3, Ying-hua SHI1,2,3, Zhen-tian LI1,2,3, Bo-shuai LIU1,2,3, Ya-lei CUI1,2,3, Zhi-chang WANG1,2,3, Yong-tao LI4, Xiao-yan ZHU1,2,3()   

  1. 1.College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450046,China
    2.Henan Key Laboratory of Innovation and Utilization of Grassland Resources,Zhengzhou 450046,China
    3.Henan Forage Engineering Technology Research Center,Zhengzhou 450046,China
    4.College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China
    5.Henan Provincial Animal Husbandry Bureau Forage and Feed Station,Zhengzhou 450002,China
  • Received:2025-07-14 Revised:2025-10-09 Online:2026-07-20 Published:2026-05-21
  • Contact: Xiao-yan ZHU

摘要:

旨在优化苜蓿皂苷的发酵和超声提取工艺。选用酿酒酵母、枯草芽孢杆菌和干酪乳杆菌为发酵菌株,在单因素试验中比较3种单菌及1∶1∶1复合菌的发酵效果,系统分析发酵时间(24、48、72、96 h)、葡萄糖添加量(0.05、0.10、0.15、0.20 g)、发酵料液比[1∶0.8、1∶1.2、1∶1.6、1∶2.0 (g∶mL)]和菌液添加量(0.04、0.06、0.08、0.10 mL)对皂苷得率的影响,并评估菌型与发酵参数的交互作用,每个处理设3个独立重复。基于结果筛选出最优菌型及关键发酵参数,采用Box-Behnken响应面法优化发酵工艺,并通过3次独立验证试验评估模型的拟合度与可重复性。进一步以最优发酵产物为原料,考察超声时间(45、60、75、90 min)、超声料液比(1∶10、1∶20、1∶30、1∶40)和超声温度(25、37、49、61 ℃)对皂苷得率的影响,分析超声参数与菌型的交互效应,建立响应面模型并优化提取工艺。结果表明:3种菌1∶1∶1组合发酵效果优于单菌发酵(P<0.05),最优发酵条件为料液比1∶1.27、发酵时间50.3 h、菌添加量0.062 mL,皂苷得率为11.68%;在发酵联合超声条件(料液比1∶23.1、超声时间72.7 min、超声温度47.3 ℃)下,皂苷得率提升至13.87%,较对照组提高98.4%(P<0.05)。本研究构建了高效、绿色的发酵-超声协同提取工艺,为苜蓿皂苷的高效提取与后续高值化开发提供了技术支持。

关键词: 苜蓿皂苷, 益生菌发酵, 超声提取, 工艺优化, 生物活性成分

Abstract:

This study aimed to optimize the fermentation and ultrasound-assisted extraction processes for alfalfa (Medicago sativa) saponins. Saccharomyces cerevisiaeBacillus subtilis, and Lactobacillus casei were selected as fermentation strains for testing. Single-factor experiments were conducted to compare three single strains and their 1∶1∶1 mixed culture fermentation, and to systematically evaluate the influence of fermentation time (24, 48, 72, and 96 hours), glucose addition (0.05, 0.10, 0.15, and 0.20 g per g of alfalfa powder), fermentation solid-to-liquid ratio [1∶0.8, 1∶1.2, 1∶1.6, and 1∶2.0 (g∶mL)], and strain inoculum volume (0.04, 0.06, 0.08, and 0.10 mL) on the saponin yield. The interactions between strain type and fermentation parameters were also assessed, with three independent replicates for each treatment. Based on the results, the optimal strain type combination and key fermentation parameters were determined, and a Box-Behnken design (BBD) was applied to further optimize the fermentation conditions. Model fit and reproducibility were evaluated through three independent validation experiments. Furthermore, using the optimal fermented product as the raw material, the effects of ultrasonication time (45, 60, 75, and 90 min), ultrasonication solid-to-liquid ratio (1∶10, 1∶20, 1∶30, and 1∶40), and ultrasonication temperature (25, 37, 49, and 61 ℃) on saponin yield were investigated. The interactions between ultrasonication parameters and strain type were analyzed, and BBD was employed to optimize the extraction process. The results showed that the 1∶1∶1 mixed-culture fermentation achieved a significantly higher saponin yield than single-strain fermentation (P<0.05). The optimal fermentation conditions were a solid-to-liquid ratio of 1∶1.27, a fermentation time of 50.3 hours, and a strain inoculation volume of 0.062 mL. Under this regime the saponin yield was 11.68%. Under the combined fermentation and ultrasound-assisted extraction conditions (solid-to-liquid ratio 1∶23.1, ultrasonication time 72.7 min, and temperature 47.3 ℃), the saponin yield increased to 13.87%, which was 98.4% higher than that of the control (P<0.05). This study developed an efficient and eco-friendly fermentation-ultrasound synergistic extraction process, providing technical support for the efficient extraction and subsequent high-value utilization of alfalfa saponins.

Key words: alfalfa saponins, probiotic fermentation, ultrasonic extraction, process optimization, bioactive components