International Research Fellowship Program: Electrochemical Biosensors for the Detection of Individual Enzyme Catalysis Events

国际研究奖学金计划：用于检测单个酶催化事件的电化学生物传感器

• 批准号: 0754396
• 负责人: Edgar Goluch
• 金额: $ 14.98万
• 依托单位: Goluch Edgar D
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0754396&HistoricalAwards=false
• 关键词: International; Research; Fellowship; Program; Electrochemical

0754396GoluchThe International Research Fellowship Program enables U.S. scientists and engineers to conduct nine to twenty-four months of research abroad. The program's awards provide opportunities for joint research, and the use of unique or complementary facilities, expertise and experimental conditions abroad.This award will support a twenty-four month research fellowship by Dr. Edgar D. Goluch to work with Dr. Serge Lemay at Delft University in the Netherlands.The goal of the proposal is to develop a new technique that electrochemically detects the activity of a single enzyme in real time by providing a signal each time a product molecule is generated. The approach demonstrates a new generation of electrochemical biosensors with single molecule detection sensitivity that will be employed to study the internal dynamics of individual enzyme molecules. This technique uses a microfabricated device that consists of two electrodes in a glass channel that are separated by about 10 nanometers. The electrodes detect electrochemically active molecules that are located in the liquid-filled volume between them. Catalysis is studied by placing a single enzyme that can convert inactive substrates into redox-active products between the electrodes. A reversibly redox-active molecule can rapidly gain and lose electrons when exposed to an electrical potential. While a single electron transfer event is very difficult to detect, the nanoscale gap allows millions of electrons to be transported by a redox-active molecule every second, thus producing a measurable amount of electrical current. The dramatic amplification of the electrochemical signals provided by this detection scheme allows the construction of electrochemical biosensors with unprecedented sensitivity and selectivity. Tyrosinase, which catalyzes the conversion of phenols into redox-active quinone molecules, is used as the model system. Of particular interest are the structural changes that the enzyme undergoes over time and their effect on enzyme activity, as well as the effects of external factors, such as pH, temperature, and buffer concentration. Finally, the influence of dramatic fluctuations in substrate concentration generated by the minute fluid volume of the system is being investigated.

0754396 GOLUCHTHE国际研究奖学金计划使美国科学家和工程师能够在国外进行九至二十四个月的研究。 该计划的奖项提供了共同研究的机会，以及在国外使用独特或补充设施，专业知识和实验条件。该奖项将支持Edgar D. Goluch博士的二十四个月研究奖学金，以与荷兰Delft University的Serge Lemay博士合作，该提议的目标是开发一项新型技术，以启用一定的时间，以启用一项新的时间，以启用一定的活动，以实现一定的启动，以实现一定的时间，该产品是由一定的启用了一个活动。产生分子。 该方法证明了新一代具有单分子检测敏感性的电化学生物传感器，该生物传感器将用于研究单个酶分子的内部动力学。 该技术使用一种微型制动设备，该设备由玻璃通道中的两个电极组成，该电极被约10纳米分离。 电极检测位于它们之间液体填充体积的电化学活性分子。 通过放置可以将无活性底物转换为电极之间的氧化还原活性产物的单个酶来研究催化。 当暴露于电势时，可逆的氧化还原活性分子可以迅速获得并损失电子。 虽然很难检测到单个电子传输事件，但纳米级间隙允许每秒通过氧化还原活性分子传输数百万个电子，从而产生可测量量的电流。 通过这种检测方案提供的电化学信号的急剧放大允许建造具有前所未有的灵敏度和选择性的电化学生物传感器。 酪氨酸酶将苯酚转化为氧化还原活性喹酮分子，被用作模型系统。 特别令人感兴趣的是酶会随着时间的流逝而经历的结构变化及其对酶活性的影响以及外部因素（例如pH，温度和缓冲液浓度）的影响。 最后，正在研究系统的微小流体体积产生的底物浓度中急剧波动的影响。