物理化学学报 >> 2009, Vol. 25 >> Issue (10): 2061-2067.doi: 10.3866/PKU.WHXB20091037

研究论文 上一篇    下一篇

HRP在大孔笼状介孔分子筛FDU-12上的固定及直接电化学

马国仙, 钟庆东, 鲁雄刚, 陆天虹   

  1. 上海大学材料科学与工程学院, 上海 200072|南京师范大学化学与环境科学学院, 南京 210097
  • 收稿日期:2009-05-11 修回日期:2009-07-03 发布日期:2009-09-29
  • 通讯作者: 马国仙 E-mail:maguoxian@shu.edu.cn

Immobilization and Direct Electrochemistry of Horseradish Peroxidase on Large Cage-Like Mesoporous Silica FDU-12

MA Guo-Xian, ZHONG Qing-Dong, LU Xiong-Gang, LU Tian-Hong   

  1. School of Materials Science and Engineering, Shanghai University, Shanghai 200072, P. R. China|College of Chemistry and Environment Science, Nanjing Normal University, Nanjing 210097, P. R. China
  • Received:2009-05-11 Revised:2009-07-03 Published:2009-09-29
  • Contact: MA Guo-Xian E-mail:maguoxian@shu.edu.cn

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摘要：

用吸附的方法将辣根过氧化物酶(HRP)固定到三维笼状介孔分子筛FDU-12中, 傅立叶变换红外光谱(FTIR)和电化学交流阻抗谱结果表明, 固定后的HRP没有变性, 并表现出良好的直接电化学性质, 其式量电位(E0')为-0.325 V, 在40-300 mV·s-1范围内, 它不随扫描速率变化而变化. 电化学反应速率常数(ks)为1.200 s-1. 固定后的HRP对H2O2有稳定的电催化活性, 该固定酶的方法具有简单、易操作和电极稳定性良好等优点, 可用于获得其他酶或氧化还原蛋白质的直接电子转移以及第三代生物传感器电极的制备.

关键词: 辣根过氧化物酶, FDU-12, 直接电化学, 过氧化氢, 生物传感器

Abstract:

We report on a direct electron transfer and bioelectrocatalysis of horseradish peroxidase (HRP) immobilized in mesoporous silica FDU-12 with three-dimensional (3D) large cages and entrances. UV-Vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and impedance spectroscopy were used to prove the interaction between HRP and FDU-12. Results from cyclic voltammetry show that immobilized HRP can undergo a direct quasi-reversible electrochemical reaction. Its formal potential, E0', is -0.325 V in a phosphate buffer solution (PBS, pH 6.9) and this is almost independent of the scan rate from 40 to 300 mV·s -1. The experimental results also demonstrate that the immobilized HRP retains its bioelectrocatalytic activity for the reduction of H2O2. Furthermore, the electrode can be stored at 4 °C for several weeks without any loss of enzyme activity. Thus, FDU-12 is a novel matrix for realizing a direct electron transfer of various proteins and enzymes and the preparation of electrodes for the third biosensors.

Key words: Horseradish peroxidase, FDU-12, Direct electrochemisrty, Hydrogen peroxide, Biosensor