黑曲霉β-葡萄糖苷酶An3987酶法制备白藜芦醇和宝霍苷I

Enzymatic Preparation of Resveratrol and Baohuoside I by β-Glucosidase An3987 from Aspergillus niger

摘要:

β-葡萄糖苷酶在酶法制备白藜芦醇和宝霍苷I中具有重要作用。为了挖掘具有高效转化效率和对乙醇具有较好耐受性的酶资源，研究了黑曲霉CBS 513.88的β-葡萄糖苷酶An3987（XP_001394024.1）对白藜芦醇和宝霍苷I的转化率，乙醇耐受性以及pH值和温度稳定性，单因素优化其水解虎杖苷和淫羊藿苷制备白藜芦醇和宝霍苷I。研究表明β-葡萄糖苷酶An3987对虎杖苷和淫羊藿苷的酶活分别为130.02 IU/mL和70.37 IU/mL；具有较好的乙醇耐受性，10%的乙醇体积分数下对酶活力基本无影响，40%的乙醇体积分数下其仍能够保持45%的酶活力；最适温度和pH值分别为65 ℃和4.5。应用An3987水解虎杖苷制备白藜芦醇，65 ℃条件下反应时间20 min转化率为90.43%；水解淫羊藿苷制备宝霍苷I，60 ℃条件下反应时间40 min转化率为92.26%。该研究为黑曲霉β-葡萄糖苷酶制备白藜芦醇和宝霍苷I提供了理论基础。

Abstract:

β-Glucosidase plays an important role in the enzymatic preparation of resveratrol and baohuoside I. In order to obtain the enzyme resources with high conversion efficiency and good ethanol tolerance, β-glucosidase An3987 (XP_001394024.1) was selected from Aspergillus niger CBS 513.88 to study its conversion rate between resveratrol and baohuoside I, ethanol tolerance, pH and temperature stability, and its hydrolysis of polydatin and icariin was optimized by Dsingle factor experiments to prepare resveratrol and baohuoside I. The results showed that the activities of β-glucosidase An3987 against polydatin and icariin were 130.02 IU/mL and 70.37 IU/mL, respectively; An3987 possessed good ethanol tolerance, with 10% (v/v) ethanol having no effect on the enzyme activity of An3987, and 40% (v/v) ethanol still letting 45% enzyme activity retained; The optimum temperature and pH were 65 ℃ and pH 4.5. Using An3987 to hydrolyze polydatin to prepare resveratrol at 65 ℃ for 20 min, the conversion rate was 90.43%; the hydrolysis of icariin at 60 ℃ for 40 min, the conversion rate for An3987 was 92.26%. This study provides a theoretical basis for the preparation of resveratrol and baohuoside I by the β-glucosidase from Aspergillus niger.