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  

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

作　　者：姚轶俊 王立峰[1] 鞠兴荣 

机构地区：[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　　处：《食品科学》2017年第24期177-182,共6页Food Science

基　　金：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).

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 mgβ-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 cm^3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。This study was conducted to evaluate the feasibility of conversion of isoflavones in soymilk using celluloseimmobilized β-glucosidase.β-glucosidase was immobilized onto cellulose particles with a loading capacity of over 40 mg of enzyme/40 cm^3 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 cm^3 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.

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 

分 类 号：Q814.2[生物学—生物工程]