肌红蛋白在海藻酸钠水凝胶中的电化学和电催化特性

海藻酸钠(Sodium Alginate,SA)是由L-葡萄糖醛酸与D-甘露糖醛酸组成的高分子线性糖醛酸,常作为固定化酶包埋材料。本文研究了海藻酸钠水凝胶膜中的肌红蛋白在磷酸盐缓冲溶液中的直接电化学和酶催化性质,探讨了测定H2O2和NO2^-的可能性。     

海藻酸钠水凝胶固定化血红蛋白的电化学性质

用海藻酸钠(SA)将血红蛋白(Hb)固定在热裂解石墨电极表面，制备了Hb—SA膜修饰电极。包埋在SA膜中的血红蛋白与电极直接传递电子。在pH7．0的磷酸盐缓冲溶液和磷酸盐／乙醇混合溶液中均可得到一对可逆氧化还原峰，这是血红蛋白辅基血红素Fe^3 ／Fe^2 电对的氧化还原。电势随溶液pH值增加而负移且呈线性关系，直线斜率为-41．0mV／pH，说明血红蛋白的电子传递过程伴随有质子的转移。并研究了Hb—SA膜修饰电极对O2、H2O2、NO和六氯乙烷(HCE)的电催化性质。     

辣根过氧化物酶在海藻酸钠水凝胶中的电化学和电催化特性

辣根过氧化物酶在海藻酸钠水凝胶中的电化学和电催化特性;辣根过氧化物酶;海藻酸钠;直接电化学电;催化     

血红蛋白在有机溶剂中的电化学行为及其类酶电催化研究

本文研究了Hb/Sol-gel膜修饰碳糊电极在有机溶剂介质中的直接电化学和类过氧化物酶电催化特性。发现在以甲醇、正内醇和乙腈介质中,Hb修饰电极能够实现其与电极之间的直接电子转移,并且电子转移的动力学过程与水溶液体系相似。然而,在这些有机溶剂介质中,电子转移的可逆性不如水介质中好,同时氧化还原峰电位差均不大于水溶液体系,表明有机溶剂对Hb的直接电子转移产生抑制作用。Hb/Sol-gel修饰电极在有机溶剂介质中和水介质中一样,对其特异性底物H2O2产生过氧化物酶的活性,但其活性均不如水溶液体系强。     

纳米硫化镉修饰玻碳电极对血红蛋白的直接电化学和生物电催化

0IntroductionStudies of direct electrochemistry of proteins orenzymes at electrodes can serve as a basis for build-ing electrochemical biosensors,enzymatic bioreactors,and biomedical devices[1].This approach simplifiessuch devices without using mediators and is of partic-ular significance for fabricating the third generationbiosensors[2].For example,if a protein or enzyme im-mobilized on electrode surface is capable of directelectron transfer without loss of bioactivities,it can beused in the …     

血红蛋白在碳纳米管修饰碳糊电极上的直接电化学行为

利用吸附法将血红蛋白(Hb)固定在碳纳米管修饰碳糊电极表面,制成稳定的固载Hb碳纳米管修饰电极,研究了Hb在碳纳米管修饰电极上的直接电化学行为.固载Hb的碳纳米管修饰电极在pH=7.0的PBS（磷酸盐缓冲溶液）中有一对相当可逆的循环伏安氧化还原峰,为Hb血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.式电位为－0.160 V(vs SCE),随扫描速度变化很小.电子转移数为1.021,近似为一个辅基发生电子转移.Hb在碳纳米管修饰电极表面的电子转移常数为0.0816 s－1,远大于亚甲蓝作媒介体时Hb的电子转移反应速率常数.应用于过氧化氢、三氯乙酸和硝基苯等的电催化还原,固定在碳纳米管修饰碳糊电极的血红蛋白表现出稳定且较高的催化活性.     

葡萄糖氧化酶在壳聚糖包埋单壁碳纳米管膜中的直接电化学与电催化研究

本文以葡萄糖氧化酶(GOx)为研究对象,采用壳聚糖(Chitosan)-单壁碳纳米管(SWCNTs)复合膜固定GOx构建一种膜体系,研究了GOx在膜上的直接电子转移和对葡萄糖的电催化.通过循环伏安法,在pH=7.0的磷酸盐缓冲溶液中对膜电极进行表征,得到一对形态良好的可逆氧化还原峰,探讨了在不同pH值的磷酸盐缓冲溶液及...     

血红蛋白在壳聚糖修饰碳纳米管上的电化学特性及对过氧化氢的电催化分析

用壳聚糖对多壁碳纳米管进行修饰,构建了一种用于固定血红蛋白的新型复合材料,并研究了血红蛋白在该碳纳米管上的电化学性质及其对过氧化氢的电催化活性.扫描电镜结果表明,壳聚糖修饰的多壁碳纳米管呈单一的纳米管状,并能均匀分散在玻碳电极表面.紫外光谱分析表明血红蛋白在该复合膜内能很好地保持其原有的二级结构.将该材料固定在玻碳电极上后,血红蛋白能成功地实现其直接电化学.根据峰电位差随着扫描的变化,计算得到血红蛋白在壳聚糖修饰的碳纳米管膜上的电荷转移系数为0.57,表观电子转移速率常数为7.02 s-1.同时,该电极对过氧化氢显示出良好的催化性能,电流响应信号与H2O2浓度在1.0×10-6 ～1.5×10-3 mol/L间呈线性关系,检出限为5.0×10-7 mol/L.修饰电极显示了良好的稳定性.     

Hb-CTAB/GC电极的电化学行为研究及对NO的电催化检测

生物大分子的直接电化学研究是近年来研究的一个热点[1-3]。血红蛋白(Hb)是一种含血红素辅基的球型蛋白质,在生物体内起存储和运输氧气的作用,其所含的血红素辅基能发生得失电子的反应。但是,在血红蛋白分子中,一方面,血红素辅基被多肽链所包裹,使得血红素辅基不易接近普通电极     

碳糊电极上无机膜固载血红蛋白的直接电化学

报道了用硅溶胶-凝胶(Sol-gel)膜将血红蛋白(Hb)固载于碳糊电极上的直接电化学行为.研究结果表明,Hb-Sol-gel修饰的碳糊电极在pH=7.0的缓冲溶液中于-0.275V(vs.Ag/AgCl)处有一对可逆的循环伏安氧化-还原峰,为Hb血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.HbFe(Ⅲ)/Fe(Ⅱ)电对的式量电位在pH5.0～11.0范围内与溶液pH值呈线性关系,表明Hb的电化学还原很可能是一个质子伴随着一个电子的电极过程.FTIR光谱证实,Sol-gel膜对Hb的固载没有破坏其天然结构.Hb-Sol-gel修饰的碳糊电极能够催化还原H₂O₂,可望将其用于制作第三代生物传感器.     

Direct electrochemistry and electrocatalysis of hemoglobin immobilized in TiO2 nanotube films

Titanium oxide nanotubes (TiO₂-NTs) synthesized by the hydrothermal method had been prepared as the co-immobilization matrix to incorporate hemoglobin (Hb) successfully. The nanostructures of TiO₂-NTs were investigated by X-ray diffraction and high-resolution electron microscopy. The Hb immobilized in TiO₂-NTs had a similar structure to the native of Hb and retained its near-native conformations as characterized by the UV–vis and FT-IR spectroscopy. A couple of quasi-reversible redox peaks with a formal potential of −0.34 V (vs. SCE) in 0.10 M pH 7.0 phosphate buffered saline (PBS) were observed. The amperometric response of the immobilized Hb linearly to H₂O₂ concentration ranged from 4 μM to 64 μM with a detection limit of 4.637 × 10⁻⁶ M and the high stability of the immobilized Hb in TiO₂-NTs constituted a promising platform for the development of biosensors.     

琼脂糖水凝胶固定化血红素蛋白质的电化学研究

利用琼脂糖（agarose）水凝胶将肌红蛋白（Mb）、血红蛋白（Hb）、辣根过氧化物酶（HRP）和过氧化氢酶（Cat）4种血红素蛋白质固定在裂解石墨电极表面,形成稳定的血红素蛋白质-agarose膜修饰电极。在agarose膜中,Mb、Hb、HRP和Cat直接与电极传递电子。4种血红素蛋白质的式电势都随溶液pH的增加而负移且呈线性关系,表明电子传递过程伴随着质子转移。     

Direct electrochemistry and electrocatalysis of hemoglobin in gelatine film modified glassy carbon electrode

Direct electrochemical and electrocatalytic behavior of hemoglobin (Hb) immobilized on glass carbon electrode (GCE) containing gelatine (Gel) films was investigated. The characteristics of Hb/Gel film modified GC electrode were performed by using SEM microscopy, UV-vis spectroscopy and electrochemical methods. The immobilized Hb showed a couple of quasi-reversible redox peak with a formal potential of −0.38 V (versus SCE) in 0.1 M pH 7.0 PBS. The formal potential changed linearly from pH 4.03 to 8.41 with a slope value of −52.0 mV pH⁻¹, which suggested that a proton transfer was accompanied with each electron transfer (ET) in the electrochemical reaction. The Hb/gelatine/GCE displayed a rapid amperometric response to the reduction of H₂O₂ and nitrite.     

Direct electrochemistry and electrocatalysis of hemoglobin entrapped in semi-interpenetrating polymer network hydrogel based on polyacrylamide and chitosan

Semi-interpenetrating polymer network (semi-IPN) hydrogel based on polyacrylamide (PAM) and chitosan was prepared to immobilize redox protein hemoglobin (Hb). The Hb-PAM-chitosan hydrogel film obtained has been investigated by scanning electron microscopy (SEM) and UV-VIS spectroscopy. UV-VIS spectroscopy showed that Hb kept its secondary structure similar to its native state in the Hb-PAM-chitosan hydrogel film. Cyclic voltammogram of Hb-PAM-chitosan film-modified glass carbon (GC) electrode showed a pair of well-defined and quasi-reversible redox peaks for Hb Fe(III)/Fe(II), indicating that direct electron transfer between Hb and GC electrode occurred. The electron-transfer rate constant was about 5.51 s(-1) in pH 7.0 buffers, and the formal potential (E degrees ') was -0.324 V (vs. SCE). The dependence of E degrees ' on solution pH indicated that one-proton transfer was coupled to each electron transfer in the direct electron-transfer reaction. Additionally, Hb in the semi-IPN hydrogel film retained its bioactivity and showed excellent electrocatalytic activity toward H(2)O(2). The electrocatalytic current values were linear with increasing concentration of H(2)O(2) in a wide range of 5-420 microM. The unique semi-IPN hydrogel would have wide potential applications in direct electrochemistry, biosensors and biocatalysis.     

Direct electrochemistry and electrocatalysis of heme proteins entrapped in agarose hydrogel films in room-temperature ionic liquids

The electrochemistry and electrocatalysis of a number of heme proteins entrapped in agarose hydrogel films in the room-temperature ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF(6)]) have been investigated. UV-vis and FTIR spectroscopy show that the heme proteins retain their native structure in agarose film. The uniform distribution of hemoglobin in agarose-dimethylformamide film was demonstrated by atomic force microscopy. Cyclic voltammetry shows that direct electron transfer between the heme proteins and glassy carbon electrode is quasi-reversible in [bmim][PF(6)]. The redox potentials for hemoglobin, myoglobin, horseradish peroxidase, cytochrome c, and catalase were found to be more negative than those in aqueous solution. The charge-transfer coefficient and the apparent electron-transfer rate constant for these heme proteins in [bmim][PF(6)] were calculated from the peak-to-peak separation as a function of scan rate. The heme proteins catalyze the electroreduction of trichloroacetic acid and tert-butyl hydroperoxide in [bmim][PF(6)]. The kinetic parameter I(max) (maximum current at saturation concentration of substrate) and the apparent K(m) (Michaelis-Menten constant) for the electrocatalytic reactions were evaluated.     

Direct electrochemistry and electrocatalysis of hydrogen peroxide using hemoglobin immobilized in hollow zirconium dioxide spheres and sodium alginate films

A biocompatible composite made from hollow zirconium dioxide microspheres (HZMS) and sodium alginate (SA) is presented and used for the construction of a biosensor for hydrogen peroxide using a gold electrode. The composition, morphology and size were studied by transmission electron microscopy. FT-IR and UV-vis spectroscopy revealed that hemoglobin (Hb) entrapped in the ZHMS retains its native structure. A pair of stable and well-defined quasi-reversible redox peaks of Hb is obtained, with a formal potential of ?0.15 V at pH 7.0. The apparent heterogeneous electron transfer rate constant is 1.15 s^?1, indicating facile electron transfer that may result from the unique nanostructures and larger surface area of the HZMS. The amperometric response of the sensor varied linearly with the concentration of hydrogen peroxide in the range from 1.75 µM to 4.9 mM, with a detection limit of 0.6 µM (at S/N?=?3). The apparent Michaelis-Menten constant $ \left( {K_{\rm{M}}^{\rm{app}}} \right) $ is 1.6 mM. The biosensor possesses high sensitivity, good reproducibility, and long-term stability.     

海藻酸钠水凝胶固载细胞色素c的蛋白膜伏安法研究

用海藻酸钠(SA)水凝胶将细胞色素c(cyt-c)固定在棱面热解石墨电极(EPPGE)表面,形成稳定的SA-cyt-c/EPPGE电极,采用蛋白膜伏安法(PFV)研究了cyt-c与电极之间的直接电化学和离子强度、pH、外加金属离子对其电化学行为的影响及其电催化性能。结果表明:cyt-c中血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对表现出准可逆行为,pH7.0、NaCl浓度为0.1 mol/L时电化学活性最高,外加金属离子则有抑制作用,SA-cyt-c/EPPGE可催化还原O2,有可能发展成为一种溶解氧的生物传感器。     

Direct electrochemistry and electrocatalysis of hemoglobin in sodium alginate film on a BMIMPF6 modified carbon paste electrode

A room temperature ionic liquid (RTIL) modified carbon paste electrode was constructed based on the substitute of paraffin with 1-butyl-3-methyl-imidazolium hexafluorophosphate (BMIMPF₆) as binder for carbon paste. Direct electrochemistry and electrocatalytic behaviors of hemoglobin (Hb) entrapped in the sodium alginate (SA) hydrogel film on the surface of this carbon ionic liquid electrode (CILE) were investigated. The presence of IL in the CILE increased the electron transfer rate and provided a biocompatible interface. Hb remained its bioactivity on the surface of CILE and the SA/Hb modified electrode showed a pair of well-defined, quasi-reversible cyclic voltammetric peaks with the apparent standard potential (E^0′) at about −0.344 V (vs. SCE) in pH 7.0 Britton–Robinson (B–R) buffer solution, which was attributed to the Hb Fe(III)/Fe(II) redox couple. UV–Vis absorption spectra indicated that heme microenvironment of Hb in SA film was similar to its native status. Hb showed a thin-layer electrochemical behavior in the SA film with the direct electron transfer achieved on CILE without the help of electron mediator. Electrochemical investigation indicated that Hb took place one proton with one electron electrode process and the average surface coverage of Hb in the SA film was 3.2 × 10⁻¹⁰ mol/cm². The immobilized Hb showed excellent electrocatalytic responses to the reduction of H₂O₂ and nitrite.