高等学校化学学报

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铁氨基黏土-葡萄糖氧化酶纳米复合催化剂的构筑及多酶级联反应研究

李柳1, 孙仕勇1(), 吕瑞1, GOLUBEVYevgeny Aleksandrovich2, 王可1, 董发勤1, 段涛3, KOTOVAOlga Borisovna2, KOTOVAElena Leonidovna4   

  1. 1.西南科技大学环境与资源学院, 绵阳 621000
    2.俄罗斯科学院乌拉尔分院科米科学中心尤什金地质研究所, 瑟克特夫卡尔 167982
    3.西南科技大学国防科学技术学院, 绵阳 621000
    4.圣彼得堡国立矿业大学, 圣彼得堡 199106
  • 收稿日期:2020-08-31
  • 通讯作者: 孙仕勇 E-mail:shysun@swust.edu.cn
  • 基金资助:
    国家自然科学基金(批准号(42011530085);41672039)、 俄罗斯基础研究基金(20-55-53019);四川省杰出青年科技基金人才培育项目(2019JDJQ0056)

Construction of Fe-aminoclay-glucose Oxidase Nanocomposite Catalyst and Its Multi-enzyme Cascade Analysis

LI Liu1, SUN Shiyong1(), LYU Rui1, GOLUBEV Yevgeny Aleksandrovich2, WANG Ke1, DONG Faqin1, DUAN Tao3, KOTOVA Olga Borisovna2, KOTOVA Elena Leonidovna4   

  1. 1.School of Environment and Resource,Southwest University of Science and Technology,Mianyang 621000,China
    2.Yushkin’s Instiute of Geology,Komi Science Center,Ural Branch of RAS,Syktyvkar 167982,Russia
    3.School of National Defence Science & Technology,Southwest University of Science and Technology,Mianyang 621000,China
    4.St. Petersburg Mining State University,St. Petersburg 199106,Russia
  • Received:2020-08-31
  • Contact: SUN Shiyong E-mail:shysun@swust.edu.cn
  • Supported by:
    ? Supported by the National Natural Science Foundation of China(42011530085);the Russian Foundation for Basic Research(20?55?53019);the Youth Science and Technology Foundation of Sichuan Province, China(2019JDJQ0056)

摘要:

以铁氨基黏土(FeAC)为载体, 通过共价交联固定葡萄糖氧化酶(GOx), 构筑了铁氨基黏土-葡萄糖氧化酶纳米复合催化剂(FeAC-GOx). 利用FeAC的过氧化物酶活性, 与GOx结合进行级联反应, 可催化葡萄糖转化为过氧化氢并产生显色反应; 采用扫描电子显微镜(SEM)、 X射线衍射(XRD)和傅里叶变换红外光谱(FTIR)对FeAC-GOx进行了形貌和结构表征, 并评价了其酶动力学参数、 催化稳定性和重复使用性等. 结果表明, GOx的固定化率可达到76.4%, 所构筑的纳米结构酶复合体系具有高效的级联催化能力. 与天然酶体系相比, FeAC-GOx具有更优异的温度和pH耐受性, 且在重复使用6次后, 酶催化活性无明显降低. 该体系不仅为新型葡萄糖传感器的开发奠定了基础, 还为多酶级联纳米结构酶的构筑提供了新思路.

关键词: 铁氨基黏土, 葡萄糖氧化酶, 固定化酶, 级联反应

Abstract:

Glucose oxidase(GOx) and Fe-aminoclay(FeAC) were combined to construct a mimic multi-enzyme system(FeAC-GOx) for cascade reaction. FeAC exhibit peroxidase-like activity which can catalyze H2O2 generated by GOx and trigger color reaction. The obtained FeAC-GOx was characterized by scanning electron microscope(SEM), X-ray diffraction(XRD), and fourier transform infrared spectroscopy(FTIR). The enzymatic kinetics, catalytic stability and reusability were evaluated. As a result, the immobilization efficiency of GOx could reach to 76.4%, and FeAC-GOx exhibited excellent cascade catalysis activity. Compared with the free enzyme, FeAC not only possessed a strong tolerance toward temperature and pH, but also presented excellent reusability. The catalytic activity of reused FeAC barely changed after six repetitions. This strategy may lay the foundation to the development of a new glucose biosensor, and provide a novel approach to design multi-enzyme cascade system.

Key words: Fe-aminoclay, Glucose oxidase, Immobilized enzyme, Cascade reaction

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