食品科学 ›› 2017, Vol. 38 ›› Issue (24): 177-182.doi: 10.7506/spkx1002-6630-201724028

• 工艺技术 • 上一篇    下一篇

纤维素载体固定化β-葡萄糖苷酶及其在大豆异黄酮水解中的应用

姚轶俊,王立峰,鞠兴荣   

  1. (1.南京财经大学食品科学与工程学院,江苏省现代粮食流通与安全协同创新中心,江苏?南京 210023;2.江南大学食品学院,江苏?无锡 214122)
  • 出版日期:2017-12-25 发布日期:2017-12-07
  • 基金资助:
    江苏省高校自然科学研究面上项目(15KJB550006);“十三五”国家重点研发计划重点专项(2016YFD0400201-1); 江苏省产学研前瞻性联合研究项目(BY2016009-01);江苏高校优势学科建设工程资助项目(PAPD)

Immobilization of β-Glucosidase onto Cellulose Particles for Application in Hydrolysis of Soybean Isoflavones

YAO Yijun, WANG Lifeng, JU Xingrong   

  1. (1. Collaborative Innovation Center for Modern Grain Circulation and Safety, College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210023, China; 2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China)
  • Online:2017-12-25 Published:2017-12-07

摘要: 建立纤维素载体固定化β-葡萄糖苷酶系统,用于黑豆浆中大豆异黄酮去糖基化反应,并对该催化系统进行了可行性评价。结果表明,在40?cm3纤维素颗粒上可结合超过40?mg?β-葡萄糖苷酶,并且能有效将4-硝基苯-β-D-葡糖苷酸水解成对硝基苯酚,其反应最适温度为50?℃。经动力学测试后可得其米氏常数Km值为(1.38±0.20)mmol/L。将该固定化酶系统用于黑豆浆中大豆异黄酮去糖基化,通过高效液相色谱检测其转化效率,表明该系统可以在30?min内利用40?cm3含酶载体将50?mL黑豆浆中的异黄酮全部转化为去糖基化形式。该固定化酶催化系统连续化催化稳定性实验表明,在经过15?次反应以及15?d之后,该固定化酶系统仍然可维持其60%的催化活性。

关键词: β-葡萄糖苷酶, 固定化, 纤维素颗粒, 黑豆浆, 异黄酮去糖基化

Abstract: This study was conducted to evaluate the feasibility of conversion of isoflavones in soymilk using cellulose-immobilized β-glucosidase. β-glucosidase was immobilized onto cellulose particles with a loading capacity of over 40 mg of enzyme/40 cm3 carrier. The immobilized enzyme exhibited a significant efficiency for the conversion of 4-nitrophenyl-β-D-glucuronide (p-NPG) to p-nitrophenol. The optimal temperature for the enzyme was 50 ℃ and the Michaelis constant (Km) value was determined to be (1.38 ± 0.20) mmol/L. All isoflavones in black soymilk were completely converted to deglycosylated derivatives within 30 min using 40 cm3 of the immobilized enzyme as analyzed by high-performance liquid chromatography (HPLC). Experimental evaluation of the stability showed that immobilized β-glucosidase retained 60% of its original activity after 15 times repeated use and after 15 days of storage.

Key words: β-glucosidase, immobilization, cellulose particles, black soymilk, isoflavone deglycosylation

中图分类号: