磁性硅基壳聚糖微球固定化柚苷酶水解柚皮苷  

Study on Hydrolysis of Naringin by Immobilized Naringinase on Magnetic Silica Chitosan Microspheres

作　　者：徐杰 孙西同 李佥[1] 罗健 李坤峰[1] 石峰 田晶[1] 费旭[2] 徐龙权[2] XU Jie;SUN Xi-tong;LI Qian;LUO Jian;LI Kun-feng;SHI Feng;TIAN Jing;FEI Xu;XU Long-quan(School of Biological Engineering,Dalian Polytechnic University,Dalian 116034,Liaoning,China;Instrumental Analysis Center,Dalian Polytechnic University,Dalian 116034,Liaoning,China;Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,Liaoning,China)

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034 [2]大连工业大学分析中心,辽宁大连116034 [3]中国科学院大连化学物理研究所,辽宁大连116023

出　　处：《精细化工》2018年第11期1885-1892,共8页Fine Chemicals

基　　金：国家自然科学基金项目(31601411);辽宁省教育厅科学研究一般项目(L2015045)~~

摘　　要：在磁性Fe3O4外包覆一层SiO2,再在其外包裹壳聚糖制备出磁性硅基壳聚糖微球（MSC）,对MSC进行环氧基修饰后用于柚苷酶的固定化研究,并对磁性硅基壳聚糖微球固定化柚苷酶水解柚皮苷的pH、温度、操作和储藏稳定性进行了考察。通过单因素实验,确定了环氧基修饰的磁性硅基壳聚糖微球（MSCE）固定化柚苷酶的最佳工艺条件为：pH 3.0,温度30℃,时间4 h、给酶量57.48 U/mL。在该条件下,MSCE固定化柚苷酶的载酶率、酶活回收率和酶比活力分别为31.29%、88.92%和409.33 U/g。与游离柚苷酶相比,MSCE固定化柚苷酶用于水解柚皮苷具有良好的pH稳定性和温度稳定性,重复使用7次后仍具有53.36%的相对酶活力,4℃条件贮存30 d后仍具有80.97%的相对酶活力。Magnetic silica chitosan microspheres（MSC） were prepared by orderly grafting SiO2 and chitosan layer on the surface of Fe3O4 nanoparticles, and after modified with epoxy groups, the obtained microspheres（MSCE） were used for immobilization of naringinase. The pH, temperature, storage and operational stability of naringin hydrolysis by the immobilized naringinase were investigated. The optimal immobilization conditions were determined as follows： immobilization pH 3.0, immobilization temperature 30 ℃, immobilization time 4 h, and crude naringinase broth activity 57.48 U/m L. Under these immobilization conditions, the binding efficiency, activity recovery rate and specific activity of the immobilized naringinase were 31.29%, 88.92% and 409.33 U/g, respectively. Comparing with free naringinase, the immobilized naringinase used for naringin hydrolysis showed better pH stability and thermal stability, and it retained 53.36% residual activity after seven consecutive operations and 80.97% residual activity after one month of storage.

关 键 词：磁性材料 壳聚糖 生物高分子 固定化 柚苷酶 生物工程 

分 类 号：Q556[生物学—生物化学]