Page 99 - 201811

第 35 卷第 11 期 精 细 化 工 Vol.35, No.11
2018 年 11 月 FINE CHEMICALS Nov. 2018

生物工程
磁性硅基壳聚糖微球固定化柚苷酶水解柚皮苷

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徐 杰 ，孙西同 ，李 佥 ，罗 健 ，李坤峰 ，
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石 峰 ，田 晶 ，费 旭 ，徐龙权
（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）
摘要：在磁性 Fe 3 O 4 外包覆一层 SiO 2 ，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。
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
中图分类号：Q556 文献标识码：A 文章编号：1003-5214 (2018) 11-1885-08

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

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XU Jie , SUN Xi-tong , LI Qian , LUO Jian , LI Kun-feng , SHI Feng ,
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TIAN Jing , FEI Xu , XU Long-quan
（1. School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, Liaoning, China; 2. Instrumental
Analysis Center, Dalian Polytechnic University, Dalian 116034, Liaoning, China; 3. Dalian Institute of Chemical
Physics, Chinese Academy of Sciences, Dalian 116023, Liaoning, China）
Abstract: Magnetic silica chitosan microspheres (MSC) were prepared by orderly grafting SiO 2 and
chitosan layer on the surface of Fe 3O 4 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/mL. 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.
Key words: magnetic materials; chitosan; biopolymer; immobilization; naringinase; biological engineering
Foundation items: National Natural Science Foundation of China (31601411); General Research Project of
Liaoning Provincial Department of Education (L2015045)

收稿日期：2017-11-07; 定用日期：2018-02-12; DOI: 10.13550/j.jxhg.20170909
基金项目：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）
作者简介：徐 杰（1991—），男，硕士生。联系人：李 佥（1982—），女，讲师，电话：0411-86323646，E-mail：liqian19820903@163.com。