Cao Lihua, Ye Jian, Tong Lili, Tang Bo
College of Chemistry, Chemical Engineering and Materials Science, Ministry of Education, Shandong Normal University, Jinan 250014 (China).
Chemistry. 2008;14(31):9633-40. doi: 10.1002/chem.200800681.
A new complex consisting of CdTe quantum dots (QDs) and glucose oxidase (GOx) has been facilely assembled to achieve considerably enhanced enzymatic activity and a wide active temperature range of GOx; these characteristics are attributed to the conformational changes of GOx during assembly. The obtained complex can be simultaneously used as a nanosensor for the detection of glucose with high sensitivity. A mechanism is put forward based on the fluorescence quenching of CdTe QDs, which is caused by the hydrogen peroxide (H2O2) that is produced from the GOx-catalyzed oxidation of glucose. When H2O2 gets to the surface of the CdTe QDs, the electron-transfer reaction happens immediately and H2O2 is reduced to O2, which lies in electron hole traps on CdTe QDs and can be used as a good acceptor, thus forming the nonfluorescent CdTe QDs anion. The produced O2 can further participate in the catalyzed reaction of GOx, forming a cyclic electron-transfer mechanism of glucose oxidation, which is favorable for the whole reaction system. The value of the Michaelis-Menton constant of GOx is estimated to be 0.45 mM L(-1), which shows the considerably enhanced enzymatic activity measured by far. In addition, the GOx enzyme conjugated on the CdTe QDs possesses better thermal stability at 20-80 degrees C and keeps the maximum activity in the wide range of 40-50 degrees C. Moreover, the simply assembled complex as a nanosensor can sensitively determine glucose in the wide concentration range from micro- to millimolar with the detection limit of 0.10 microM, which could be used for the direct detection of low levels of glucose in biological systems. Therefore, the established method could provide an approach for the assembly of CdTe QDs with other redox enzymes, to realize enhanced enzymatic activity, and to further the design of novel nanosensors applied in biological systems in the future.
一种由碲化镉量子点(QDs)和葡萄糖氧化酶(GOx)组成的新型复合物已被轻松组装成功,实现了GOx显著增强的酶活性和较宽的活性温度范围;这些特性归因于组装过程中GOx的构象变化。所获得的复合物可同时用作高灵敏度检测葡萄糖的纳米传感器。基于碲化镉量子点的荧光猝灭提出了一种机制,这种猝灭是由GOx催化葡萄糖氧化产生的过氧化氢(H2O2)引起的。当H2O2到达碲化镉量子点表面时,电子转移反应立即发生,H2O2被还原为O2,O2存在于碲化镉量子点上的电子空穴陷阱中,可作为良好的受体,从而形成非荧光的碲化镉量子点阴离子。产生的O2可进一步参与GOx的催化反应,形成葡萄糖氧化的循环电子转移机制,这对整个反应体系有利。GOx的米氏常数估计为0.45 mM L(-1),这表明到目前为止所测得的酶活性显著增强。此外,共轭在碲化镉量子点上的GOx酶在20 - 80℃具有更好的热稳定性,并且在40 - 50℃的宽范围内保持最大活性。此外,这种简单组装的复合物作为纳米传感器可以在从微摩尔到毫摩尔的宽浓度范围内灵敏地测定葡萄糖,检测限为0.10 microM,可用于直接检测生物体系中低水平的葡萄糖。因此,所建立的方法可为碲化镉量子点与其他氧化还原酶的组装提供一种途径,以实现增强的酶活性,并在未来进一步设计应用于生物体系的新型纳米传感器。