细胞色素c在植物凝胶-离子液体复合膜修饰电极上的直接电化学

用植物凝胶（GG）和亲水性离子液体（ILs）氯化1-庚基-3-甲基咪唑（[HMIM]Cl）制得一种有效的复合膜（GG-ILs）.将细胞色素c（Cyt c）包埋在该复合膜中并修饰于玻碳电极（GCE）表面,构置了细胞色素c修饰玻碳电极（GG-Cyt c-ILs/GCE）.紫外-可见光谱表明,复合膜内的Cyt c仍保持其原始...     

细胞色素c在纳米氧化铝模板修饰电极上的直接电化学

细胞色素c(Cytochrome c,Cyt c)是生物体中最常见的氧化一还原蛋白质,研究其在电极上的直接电化学,对于理解和认识生命体内的电子转移机制具有重要意义。Cytc与裸固体电极表面的直接接触通常会使其失去生物活性,因此,Cytc的电化学研究常借助于媒介体以实现其与电极之间的电子转移。纳米金属氧化物模板的表面积大且化学和光化学性质稳定,被广泛应用于太阳能电池和金属沉积等领域,本文研究氧化铝(AAO)模板对4,4’-二硫二吡啶存在下Cytc直接电化学促进作用。     

细胞色素c在自组装纳米膜电极上的有效固定及直接电化学

采用自组装方法将壳聚糖-纳米金(Chi-Nano Au)修饰到金(Au)电极上,并经进一步自组装细胞色素c(Cyt c),制得自组装膜电极Cyt c/Chi-Nano Au/Au.测定了自组装膜电极的循环伏安曲线(CV)及稳定性.结果表明,利用自组装膜电极Chi-Nano Au/Au可以有效地固定Cyt c,并实现直接电子转移反应.Cyt c在0.13～0.28V(vs Ag/AgCl)之间显示一对明显的可逆氧化还原峰;峰电流与扫描速度呈现良好的线性关系,线性方程为Ipc=0.063 64+0.003 51υ,线性相关系数为r=0.997 2,这表明该电极过程受吸附控制.此外,所制备的膜电极稳定性良好.     

细胞色素c在硒代胱氨酸修饰电极上的直接电化学

采用电化学和接触角实验方法研究了硒代胱氨酸自组装膜修饰金电极(SeCys SAMs/Au)和十六烷基三甲基溴化铵(CTAB)-硒代胱氨酸自组装复合膜修饰金电极(CTAB-SeCys SAMs/Au)的特性. 探讨了细胞色素c(Cyt c)在SeCys SAMs/Au电极和CTAB-SeCys SAMs/Au电极上的电化学行为. 实验证明SeCys可促进Cyt c在电极上的氧化还原反应, 加入CTAB后其与SeCys之间的协同作用可在Cyt c与电极之间形成一个开放的通道, 促进作用更加明显, 且在一定浓度范围内, 随CTAB浓度(1×10^-5-1×10^-4 mol·L^-1)的增大, Cyt c在CTAB-SeCys SAMs/Au电极上的氧化还原电流增大, 在接近临界胶束浓度处出现极大值. 在CTAB-SeCys SAMs/Au电极上Cyt c产生一对氧化还原峰, 其峰电位分别为0.305和0.235 V, 其电化学过程受扩散控制. 光谱实验证实SeCys对Cyt c电化学过程的促进作用是由于SeCys与Cyt c中赖氨酸残基的结合.     

DNA／聚（3-甲基噻吩）复合膜修饰电极的制备及其应用于研究8-羟基-2＇-脱氧鸟嘌呤核苷的伏安行为及测定

首先通过电聚合方法在玻碳电极表面制备了聚（3-甲基噻吩）（P3MT）修饰膜,然后在一定电位下将DNA分子电沉积到P3MT表面,制备了DNA／（P3MT）复合膜修饰玻碳电极．研究了8-羟基-2’-脱氧鸟嘌呤核苷（8-OH-dG）在该复合膜修饰电极上的伏安行为以及扫描速度、pH值和尿酸对其伏安行为和检测的影响．实验结果表明,该复合膜修饰电极结合了P3MT和DNA两者的优点,使8-OH-dG在复合膜修饰电极上的电化学行为明显改善,而且具有很好的重现性和稳定性．在0．1mol／LpH7．0的磷酸盐缓冲溶液（PBS）中,8-OH-dG的氧化峰电流与其浓度在0．28～4．2μmol／L和4．2～19．6μmol／L两个范围内成良好的线性关系,检出限为56nmol／L（S／N=3）．该研究可以为制备HPLC或毛细管电泳电化学检测器检测8-OH—dG打下一定的基础,因此在检测尿样中8-OH-dG的研究方面具有潜在的应用价值．     

细胞色素c在纳米杂化膜修饰玻碳电极上的直接电化学

以多壁碳纳米管(MWNTs)修饰玻碳(GC)电极为基底,自组装金纳米粒子(AuNPs)及L-半胱氨酸(L-Cys)研制杂化膜修饰电极(L-Cys/AuNPs/MWNTs/GC).实验表明,该膜修饰电极在pH=7.0的KH2PO4-K2HPO4缓冲溶液中对细胞色素c(Cyt c)的直接电子转移反应具有良好的电催化作用,C...     

铂-氧化铈/聚苯胺/聚砜复合膜电极的制备及对甲醇的电催化氧化

采用电化学聚合法制备了掺杂CeO2纳米粒子的聚苯胺(PAN)/聚砜(PSF)复合膜电极,在其上电沉积铂粒子,制得了铂-氧化铈/聚苯胺/聚砜的复合膜修饰电极。 复合膜的形貌和化学组分通过冷场发射扫描电子显微镜（Cold FE-SEM）和能量散射X射线谱（EDS）进行了表征,用循环伏安法和电化学交流阻抗法考察了复合膜电极对甲醇的电催化氧化性能。 结果表明,复合膜的双层多孔结构使铂粒子与CeO2粒子在复合膜内层的多孔聚苯胺上均匀沉积,粒子平均尺寸约为80 nm;CeO2为铂质量的7%时,铂-氧化铈/聚苯胺/聚砜复合膜修饰电极对甲醇有很好的电催化氧化性能和高的稳定性。     

聚苯胺对2,5—二巯基—1,3,4—噻二唑氧化还原反应的电化学？…

研究了聚苯胺（ＰＡn）膜电极在２,５－二巯基－１,３,４－噻二唑（ＤＭcＴ）溶液中电化学处理或浸泡后的循环伏安（ＣＶ）曲线的变化规律。实验结果表明,ＰＡn膜电极在 ＤＭcＴ溶液中进行电化学处理或浸泡过程可使ＤＭcＴ进入ＰＡn膜内部与ＰＡn形成复合物。ＰＡn对ＤＭcＴ的电化学催化作用可能和二者之间形成的电子给体－受体复合物有关。该复合物的电何尝氧化还原特性不同于ＰＡn对ＤＭcＴ的电化学催化作用可能和得之间形成的     

紫细菌反应中心色素蛋白复合膜电子转移研究

采用自组装单层膜方法在金电极表面形成单分子层的2,3－双巯基丁二酸（DMSA）,聚二甲基二丙烯氯化铵（PDDA）及紫细菌（Rhodobacter Sphaeroides)反应中心色素蛋白复合体的有序复合膜,使用方波估安法研究了该蛋白复合膜的电化学行为,成功地检测到该蛋白内多电对的可逆或准可逆电子转移过程,探讨了紫细菌反应中心蛋白内容各电子受体的氧化还原电位及外加电位驱动对各受体电位的影响,同时,通过对方波信号的非线形拟合,获得了该蛋白复合膜内主要电对的电子转移速率常数及电子迁移数等相关参数。     

三维有序结构的金掺杂纳米TiO_2修饰电极用于H_2O_2的测定

在ITO玻璃表面构建了三维有序多孔结构的金掺杂纳米Ti O2薄膜(3DOM GTD/ITO),同时制备了一种细胞色素c(Cyt c)酶生物传感器(Cyt c/3DOM GTD/ITO)。通过透射电镜(TEM)、扫描电镜(SEM)对修饰电极进行表征。紫外-可见光谱实验表明吸附在GTD上的Cyt c能够保持其生物活性,二级结构未被破坏。同时研究了Cyt c在3DOM GTD/ITO修饰电极表面的直接电化学及对H2O2的电催化行为。结果显示,Cyt c在3DOM GTD/ITO修饰电极上有显著的直接电化学响应,峰电流与扫描速度呈线性关系,说明该电极过程是表面电化学控制过程。Cyt c/3DOM GTD/ITO修饰电极对H2O2具有良好的催化性能,线性范围为3.0×10-7~1.70×10-5mol/L,检出限为3.6×10-8mol/L(S/N=3),响应时间为5 s,且该修饰电极具有较好的重现性和稳定性。     

Reversible immobilization and direct electron transfer of cytochrome c on a pH-sensitive polymer interface

A pH-sensitive polymer interface has been used as a matrix for reversible immobilization of cytochrome c (Cyt c) on an Au surface through a dip-coating process. The pH-sensitive behavior of the polymer brush interface has been demonstrated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) measurements. The reversible immobilization and electron-transfer properties of Cyt c have been investigated by in situ UV/Vis spectrophotometry and CV. The results have shown that the poly(acrylic acid) (PAA) brush acted as an excellent adsorption matrix and a good accelerant for the direct electron transfer of Cyt c, which gave redox peaks with a formal potential of 40 mV versus Ag/AgCl in pH 7.6 phosphate buffer solution. The average surface coverage of Cyt c on the PAA film was about 1.7 x 10(-10) mol cm(-2), indicating a multilayer of Cyt c. The electron-transfer rate constant was calculated to be around 0.19 s(-1) according to the CV experiments. The interface was subjected to in situ attenuated total internal reflection Fourier-transform infrared (ATR-FTIR) spectroscopic analysis, in order to further confirm the immobilization of Cyt c on the surface. This polymer-protein system may have potential applications in the design of biosensors, protein separation, interfacial engineering, biomimetics, and so on.     

Direct electrochemistry behavior of cytochrome c/L-cysteine modified electrode and its electrocatalytic oxidation to nitric oxide

Cytochrome c (Cyt c) was successfully immobilized on L-cysteine modified gold electrode by multicyclic voltammetry method. The electrochemical behavior of Cyt c on the L-cysteine modified electrode was explored. In 0.10 M, pH 7.0 phosphate buffer solution (PBS), Cyt c showed a quasi-reversible electrochemical redox behavior with E(pc)=0.180 V, E(pa)=0.208 V (versus Ag/AgCl). The Cyt c/L-cysteine modified electrode gave an excellent electrocatalytic activity towards the oxidation of nitric oxide, and the catalysis currents were proportional to the nitric oxide concentration in the range of 7.0 x 10(-7) to 1.0 x 10(-5) M, the linear regression equation is I (microA)=-0.124-0.003 C(NO) (microM), with a correlation coefficient 0.996, The detection limit was 3.0 x 10(-7) M (times the ratio of signal to noise, S/N=3).     

Direct Electrochemistry of Cytochrome c on a Silica Sol‐Gel Film Modified Electrode

Dilute silica sol‐gel was simply dropped on the surface of a basal plane graphite electrode (BPGE) to form a silica sol‐gel film modified electrode. Direct electrochemical response of cytochrome c (Cyt c) on the modified electrode was observed by cyclic voltammetry (CV). The results suggested that Cyt c could be tightly adsorbed on the surface of the silica sol‐gel film modified electrode. A couple of well‐defined and nearly reversible redox peaks can be observed in a phosphate buffer solution (pH 7.0), which anodic and cathodic peak potentials were at ?0.243 and ?0.306 V (vs. Ag/AgCl), respectively. Cyt c adsorbed on the surface of silica sol‐gel film shows a remarkable electrocatalytic activity for the reduction of oxygen. Based on these, a third‐generation biosensor could be constructed to detect the concentration of oxygen in aqueous solution.     

Amperometric nitrite biosensor based on a gold electrode modified with cytochrome c on Nafion and Cu-Mg-Al layered double hydroxides

An amperometric biosensor for nitrite was prepared by immobilizing cytochrome c (Cyt c) on a gold electrode that was modified with Nafion and a Cu-Mg-Al layered double hydroxide (Cu-LDH). The Cu-LDH was characterized by Fourier transform infrared spectroscopy and powder X-ray diffraction. The UV-visible spectrum suggests that Cyt c retains its native conformation in the modified film. The direct electrochemical investigation indicated that the composite film represents a good platform for the immobilization of Cyt c as well as an excellent promoter for the electron transfer between Cyt c and the gold electrode. Moreover, the biosensor showed a remarkable bioelectrocatalytic activity for the oxidation of nitrite with a linear range from 0.75 to 123 μM. The detection limit is 2?×?10^?7 M (S/N?=?3). The biosensor was successfully applied to the determination of nitrite in food samples.     

Folding control and unfolding free energy of yeast iso-1-cytochrome c bound to layered zirconium phosphate materials monitored by surface plasmon resonance

The free energy change (Delta G degrees ) for the unfolding of immobilized yeast iso-1-cytochrome c (Cyt c) at nanoassemblies was measured by surface plasmon resonance (SPR) spectroscopy. Data show that SPR is sensitive to protein conformational changes, and protein solid interface exerts a major influence on bound protein stability. First, Cyt c was self-assembled on the Au film via the single thiol of Cys-102. Then, crystalline sheets of layered alpha-Zr(O(3)POH)(2).H(2)O (alpha-ZrP) or Zr(O(3)PCH(2)CH(2)COOH)(2).xH(2)O (alpha-ZrCEP) were adsorbed to construct alpha-ZrP/Cyt c/Au or alpha-ZrCEP/Cyt c/Au nanoassemblies. The construction of each layer was monitored by SPR, in real time, and the assemblies were further characterized by atomic force microscopy and electrochemical studies. Thermodynamic stability of the protein nanoassembly was assessed by urea-induced unfolding. Surprisingly, unfolding is reversible in all cases studied here. Stability of Cyt c in alpha-ZrP/Cyt c/Au increased by approximately 4.3 kJ/mol when compared to the unfolding free energy of Cyt c/Au assembly. In contrast, the protein stability decreased by approximately 1.5 kJ/mol for alpha-ZrCEP/Cyt c/Au layer. Thus, OH-decorated surfaces stabilized the protein whereas COOH-decorated surfaces destabilized it. These data quantitate the role of specific functional groups of the inorganic layers in controlling bound protein stability.     

Self-aggregated nanoparticles from methoxy poly(ethylene glycol)-modified chitosan: synthesis; characterization; aggregation and methotrexate release in vitro

Methoxy poly(ethylene glycol)-grafted-chitosan (mPEG-g-CS) conjugates were synthesized by formaldehyde linking method and characterized by Fourier transform infrared (FT-IR) and proton nuclear magnetic resonance (¹H-NMR). The degree of substitution (DS) of methoxy poly (ethylene glycol) (mPEG) in the mPEG-g-CS molecules determined by ¹H-NMR ranged from 19% to 42%. The critical aggregation concentration (CAC) was determined by fluorescence spectroscopy using pyrene as fluorescence probe and its value was 0.07 mg/mL in water. mPEG-g-CS formed monodisperse self-aggregated nanoparticles with a roughly spherical shape and a mean diameter of 261.9 nm were prepared by the dialysis method. mPEG-g-CS self-aggregated nanoparticles were used as carriers of poorly water-soluble anticancer drug methotrexate (MTX). MTX was physically entrapped inside mPEG-g-CS self-aggregated nanoparticles by dialysis method and the characteristics of MTX-loaded mPEG-g-CS self-aggregated nanoparticles were analyzed using dynamic laser light scattering (DLLS), transmission electron microscopy (TEM). Moreover, in vitro release behavior of MTX was also investigated and the results showed that MTX was continuously released more than 50% in 48 h.     

细胞色素c在微-液/液界面上电化学行为的研究

本文采用电化学技术,研究了细胞色素c（Cyt c）在玻璃微米管尖端处形成的微-水/1,2-二氯乙烷（W/DCE）界面上的电化学行为.选用四丁基铵四苯硼（TBAT-PB）、四丁基铵四氯代苯硼（TBATPBCl）以及四丁基铵四氟代苯硼（TBATPBF）三种不同的有机相支持电解质来研究Cyt c在W/DCE界面上的反应.在电势窗较窄的含TBATPB体系中只能够观察到吸附过程;在电势窗较宽的含TBATPBCl和TBATPBF的体系中,可以同时观察到吸附与离子转移过程.当Cyt c浓度较低时,两种过程都可以观察到;当Cyt c浓度较高时,主要是吸附.文中对这些过程的机理进行了探讨.     

基于ITO电极的细胞色素c吸附层的直接电化学

采用未经修饰的铟锡氧化物(ITO)工作电极直接探测到了细胞色素c(Cytc)吸附层的氧化还原峰,并得出了Cytc的表面浓度,随着溶液浓度从2μmo·lL-1增大到10μmo·lL-1,Cytc的表面浓度相应地从0.35×10-12mo·lcm-2增大到1.53×10-12mo·lcm-2.实验获得的表面浓度倒数与溶液浓度倒数的准线性关系说明Cytc在ITO表面的吸附基本满足Langmuir等温吸附理论.对Cytc溶液的循环伏安测试结果表明参与电极反应的Cytc包括游离分子和吸附分子,前者的贡献大于后者,电极反应主要受扩散控制并呈准可逆过程.根据Nicholson方法估算得到反应物的标准异相速率常数的平均值为1.65×10-3cm·s-1.实验结果显示在室温下放置1h后Cytc吸附层电化学活性部分丧失,在80℃下放置1h后吸附层完全失活.失活的Cytc吸附层对铁氰化钾溶液在Au电极上的电极反应具有明显的阻碍作用.