β-葡萄糖苷酶对刺梨槲皮素糖苷的生物转化工艺优化研究
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贵州大学

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国家自然科学基金项目(面上项目,重点项目,重大项目)


Effect of β-glucosidase on Biotransformation of Quercetin Glycosides in Rosa roxburghii
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Guizhou Universty

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    摘要:

    以不同来源的β-葡萄糖苷酶水解刺梨槲皮素-3-O-芸香糖苷、槲皮素-3-O-鼠李糖苷和槲皮素-3-O-葡萄糖苷,探讨提高刺梨黄酮苷元释放能力的生物转化技术途径。以槲皮素含量与糖苷转化率为指标,采用高效液相色谱法对来源于嗜酸乳杆菌、木霉和杏仁的β-葡萄糖苷酶水解3种槲皮素糖苷的转化率及槲皮素含量进行动态监测,以较优β-葡萄糖苷酶转化槲皮素糖苷的酶解时间、酶解pH值、酶解温度和酶用量为单因素,考察各因素参数独立变化对指标的影响,再以Box-Behnken 方法研究各因素及其交互作用对转化率的影响,优化工艺条件。杏仁β-葡萄糖苷酶水解3种糖苷转化所得槲皮素含量最高,对不同底物的转化率由高到低依次为槲皮素-3-O-葡萄糖苷(74.10%)、槲皮素-3-O-芸香糖苷(64.30%)、槲皮素-3-O-鼠李糖苷(31.80%)。杏仁β-葡萄糖苷酶优化水解工艺条件为水解时间28.90 min,水解pH值4.9,水解温度52 ℃,酶与底物质量比0.08 %。此条件下得到槲皮素-3-O-芸香糖苷转化率71.48 %,槲皮素-3-O-鼠李糖苷转化率36.32 %,槲皮素-3-O-葡萄糖苷转化率77.86 %。

    Abstract:

    Hydrolysing quercetin-3-O-rutinoside, quercetin-3-O-rhamnoside, and quercetin-3-O-glucoside with β-glucosidase from different sources to explore the improvement of Rosa roxburghii biotransformation technology approach of flavonoid aglycon releasing ability. Using quercetin content and glycoside conversion rate as indicators, the conversion rate of quercetin glycosides and quercetin hydrolyzed by β-glucosidase from Lactobacillus acidophilus, Trichoderma, and almond was analyzed by HPLC.The content is dynamically monitored, and the enzymatic hydrolysis time, enzymatic hydrolysis pH, enzymatic hydrolysis temperature, and enzyme dosage of the better β-glucosidase to convert quercetin glycoside are single factors.The effects of independent changes of each factor parameter on the index are examined. The Box-Behnken method was used to study the influence of each factor and its interaction on the conversion rate, and to optimize the process conditions. Almond β-glucosidase hydrolyzes the three glycosides and converts them to the highest quercetin content. The conversion rates for different substrates are in the order of quercetin-3-O-glucoside(74.10%)>quercetin-3-O-rutinoside(64.30%)>quercetin-3-O-rhamnoside (31.80%). The optimal optimized hydrolysis process conditions for almond β-glucosidase were hydrolysis time of 28.90 min, hydrolysis pH value of 4.9, hydrolysis temperature of 52 ℃, and the enzyme-substrate mass ratio of 0.08%. Under these conditions, the conversion rate of quercetin-3-O-rutinoside was 71.48%, the conversion rate of quercetin-3-O-rhamnoside was 36.32%, and the conversion rate of quercetin-3-O-glucoside was 77.86%.

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  • 收稿日期:2020-01-06
  • 最后修改日期:2020-03-25
  • 录用日期:2020-02-20
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