Structural characterization and electrocatalytic application of hemoglobin immobilized in layered double hydroxides modified with hydroxyl functionalized ionic liquid.
Structural characterization and electrocatalytic application of hemoglobin immobilized in layered double hydroxides modified with hydroxyl functionalized ionic liquid.
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DOI：
10.1016/j.jcis.2014.07.012
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发表时间：
2014-11
影响因子：
9.9
通讯作者：
Tianrong Zhan;Qi Yang;Yumei Zhang;Xinjun Wang;Jie Xu;W. Hou
中科院分区：
文献类型：
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作者：
Tianrong Zhan;Qi Yang;Yumei Zhang;Xinjun Wang;Jie Xu;W. Hou
分别通过吸附法和共沉淀法将血红蛋白（Hb）固定在羟基功能化离子液体（HFIL）修饰的Zn2Al层状双氢氧化物（LDH）中。吸附实验表明，HFIL 的存在可以提高最大蛋白质负载量。然而，共沉淀技术的 Hb 负载量远高于吸附技术。通过 XRD、FTIR、UV-vis 和荧光光谱研究了 HFIL 对 Hb 和 LDH 相互作用的作用。尽管通过共沉淀技术捕获在 HFIL-LDH 中的 Hb 四级结构（表示为 HFIL-LDH-Hbcop）可能比 LDH（LDH-Hbcop）中的四级结构发生的改变稍多，但其二级结构和氧化还原活性血红素基团保持完整。通过 SEM 和 TEM 图像分析 LDH-Hbcop 和 HFIL-LDH-Hbcopbiohybrid 的形态。固定化 Hb 的直接电化学表明共沉淀生物电极的性能优于相应的吸附生物电极。无论吸附还是共沉淀，HFIL 的引入都可以明显促进电子传输。在所有生物电极中，HFIL–LDH–Hbcop/GCE 对 H2O2 测定显示出最佳的电催化活性，具有较大的电活性 Hb 百分比 (6.76%)、较高的灵敏度 (40.63 A/M cm2) 和较低的检测限 (0.0054 μmol/L)。因此，HFIL-LDH可以通过共沉淀技术有效地固定酶，这在电化学生物传感器的制造中具有潜在的应用。
Hemoglobin (Hb) was immobilized in Zn2Al-Layered Double Hydroxides (LDH) modified with Hydroxyl Functionalized Ionic Liquid (HFIL) through adsorption and coprecipitation method, respectively. Adsorption experiments showed that the presence of HFIL could enhance the maximum protein loading. However, the Hb loading through coprecipitation technique was far higher than that for adsorption. The role of HFIL on the interaction between Hb and LDH was investigated by XRD, FTIR, UV–vis and fluorescence spectroscopies. Although the quaternary structure of Hb entrapped in HFIL–LDH through coprecipitation technique (denoted as HFIL–LDH–Hbcop) might be altered slightly more than that in LDH (LDH–Hbcop), its secondary structure and redox-active heme groups kept intact. Morphologies of LDH–Hbcopand HFIL–LDH–Hbcopbiohybrids were analyzed through SEM and TEM images. The direct electrochemistry of the immobilized Hb indicated that the coprecipitation bioelectrode performed better than that of the corresponding adsorption one. Regardless of adsorption and coprecipitation, the introduction of HFIL could distinctly promote the electron transport. Among all bioelectrodes, HFIL–LDH–Hbcop/GCE displayed the best electrocatalytic activity for H2O2determination with a larger electroactive Hb percentage (6.76%), higher sensitivity (40.63 A/M cm2) and lower detection limit (0.0054 μmol/L). So HFIL–LDH could effectively immobilize enzymes via coprecipitation technique, which had potential applications in the fabrication of electrochemical biosensors.