Enzyme based electrochemical biosensors are divided into three generations according to their type of electron transfer from the cofactors of the enzymes to the electrodes. Although the 3rd generation sensors using direct electron transfer (DET) type enzymes are ideal, the number of enzyme types which possess DET ability is limited. In this study, we report of a glucose sensor using mediator-modified glucose dehydrogenase (GDH), that was fabricated by a new quick-and-easy method using the pre-functionalized amine reactive phenazine ethosulfate (arPES). Thus mediator-modified GDH obtained the ability to transfer electrons to bulky electron acceptors as well as electrodes. The concentration of glucose was successfully measured using electrodes with immobilized PES-modified GDH, without addition of external electron mediators. Therefore, continuous monitoring systems can be developed based on this “2.5th generation” electron transfer principle utilizing quasi-DET. Furthermore, we successfully modified two other diagnostically relevant enzymes, glucoside 3-dehydrogenase and lactate oxidase, with PES. Therefore, various kinds of diagnostic enzymes can achieve quasi-DET ability simply by modification with arPES, suggesting that continuous monitoring systems based on the 2.5th generation principle can be developed for various target molecules.

基于酶的电化学生物传感器根据其从酶的辅因子到电极的电子转移类型分为三代。尽管使用直接电子传输（DET）型酶的第三代传感器是理想的，但是具有DET能力的酶类型的数量受到限制。在这项研究中，我们报道了一种使用介体修饰的葡萄糖脱氢酶（GDH）的葡萄糖传感器，该葡萄糖传感器是通过使用预先功能化的胺反应性吩嗪乙硫酸盐（arPES）的新型快速简便方法制造的。因此，介体修饰的GDH获得了将电子转移到庞大的电子受体以及电极的能力。使用固定有PES修饰的GDH的电极成功地测量了葡萄糖的浓度，而无需添加外部电子介体。所以，可以基于准DET的“ 2.5代”电子传输原理开发连续监测系统。此外，我们成功地用PES修饰了其他两种诊断相关的酶，即糖苷3-脱氢酶和乳酸氧化酶。因此，各种诊断酶只需通过arPES修饰即可达到准DET能力，这表明可以为各种靶分子开发基于第2.5代原理的连续监测系统。