Carl von Ossietzky University of Oldenburg , School of Mathematics and Sciences, Center of Interface Science (CIS), Department of Chemistry, D-26111 Oldenburg, Germany.
Langmuir. 2013 Dec 3;29(48):15090-9. doi: 10.1021/la402561g. Epub 2013 Nov 19.
Multienzyme microstructures of glucose oxidase (GOx) and horseradish peroxidase (HRP) were prepared by layer-by-layer deposition inside microfluidic networks on glass substrates in order to allow both site-specific deposition and control of the amount of immobilized enzymes. The obtained microstructures were characterized by scanning force microscopy for the topography of the deposited layers. The local enzyme activity was characterized by the substrate-generation/tip-collection mode and the enzyme-mediated feedback mode of the scanning electrochemical microscope (SECM). These measurements provided quantitative information about the immobilized enzyme activity as a basis for adjusting enzyme loading for multienzyme structures that realize logical operations based on enzymatic conversions. Information about local HRP activity can also be obtained by optical readout using an Amplex UltraRed fluorgenic substrate and reading with a confocal laser scanning microscope with a much higher repetition rate for image acquisition. Using these principles, a layout with HRP and GOx microstructures was realized that showed the functionality of an OR Boolean logic switch.
通过在玻璃基底的微流控网络内层层沉积葡萄糖氧化酶 (GOx) 和辣根过氧化物酶 (HRP) 的多酶微结构，实现了定点沉积和固定化酶量的控制。利用扫描力显微镜对沉积层的形貌进行了表征。利用扫描电化学显微镜 (SECM) 的底物生成/尖端收集模式和酶介导的反馈模式对局部酶活性进行了表征。这些测量提供了关于固定化酶活性的定量信息，为基于酶转化实现逻辑操作的多酶结构调整酶载量提供了依据。通过使用 Amplex UltraRed 荧光底物进行光学读取，并使用具有更高重复率的共聚焦激光扫描显微镜进行图像采集，也可以获得有关局部 HRP 活性的信息。利用这些原理，实现了具有 HRP 和 GOx 微结构的布局，展示了 OR 布尔逻辑开关的功能。
