丙酮酸氧化酶在电极中的应用任务书
2020-06-26 19:50:03
1. 毕业设计(论文)的内容和要求
内容: 丙酮酸是糖酵解过程中的重要产物,是参与整个生物体基本代谢的中间产物之一,可通过乙酰coa和三羧酸循环实现体内糖、脂肪和氨基酸间的互相转化,与微生物的生长代谢有密切关系。
因此,丙酮酸在三大营养物质的代谢联系中起着重要的枢纽作用。
我们通过培养大肠杆菌,分时取样,记录了细菌生长变化曲线,并采用了高效液相色谱技术检测了发酵期间丙酮酸的浓度变化,以此来观察丙酮酸的浓度变化是否可以作为反应大肠杆菌的生理指标变化的数据之一。
2. 参考文献
[1]Akyilmaz E, Yorganci E. Construction of an amperometric pyruvate oxidase enzyme electrode for determination of pyruvate and phosphate[J]. Electrochimica Acta, 2007, 52(28):7972-7977. [2]Akyilmaz E, Yorganci E. A novel biosensor based on activation effect of thiamine on the activity of pyruvate oxidase.[J]. Biosensors Bioelectronics, 2008, 23(12):1874-1877. [3]Arai G, Noma T, Habu H, et al. Pyruvate sensor based on pyruvate oxidase immobilized in a poly(mercapto- p -benzoquinone) film[J]. Journal of Electroanalytical Chemistry, 1999, 464(2):143-148. [4]Bergmann W, Rudolph R, Spohn U. A bienzyme modified carbon paste electrode for amperometric detection of pyruvate[J]. Analytica Chimica Acta, 1999, 394(2):233-241. [5]Gajovic N, Haberm#252;ller K, Warsinke A, et al. A Pyruvate Oxidase Electrode Based on an Electrochemically Deposited Redox Polymer[J]. Electroanalysis, 2015, 11(18):1377-1383. [6]Ghica M, Brett C. Development of Novel Glucose and Pyruvate Biosensors at Poly(Neutral Red) Modified Carbon Film Electrodes. Application to Natural Samples[J]. Electroanalysis, 2010, 18(8):748-756. [7]IzumiKubo, MasakoInagawa, TakakoSugawara, et al. Phosphate Sensor Composed from Immobilized Pyruvate Oxidase and an Oxygen Electrode[J]. Analytical Letters, 2006, 24(10):1711-1727. [8]Kwan R C, Leung H F, Hon P Y, et al. Amperometric biosensor for determining human salivary phosphate.[J]. Analytical Biochemistry, 2005, 343(2):263. [9] Mak W C, Chan C, Barford J, et al. Biosensor for rapid phosphate monitoring in a sequencing batch reactor (SBR) system.[J]. Biosensors Bioelectronics, 2003, 19(3):233. [10]Mascini M, Mazzei F. Amperometric sensor for pyruvate with immobilized pyruvate oxidase[J]. Analytica Chimica Acta, 1987, 192(1):9-16. [11]Mizutani F, Tsuda K, Karube I, et al. Determination of glutamate pyruvate transaminase and pyruvate with an amperometric pyruvate oxidase sensor[J]. Analytica Chimica Acta, 1980, 118(1):65-71. [12] Mizutani F, Yabuki S, Sato Y, et al. Amperometric determination of pyruvate, phosphate and urea using enzyme electrodes based on pyruvate oxidase-containing poly(vinyl alcohol)/polyion complex-bilayer membrane[J]. Electrochimica Acta, 2000, 45(18):2945-2952. [13]金利通, 叶建山. Nafion#8212;MV修饰电极丙酮酸氧化酶生物传感器测定GPT的研究[J]. 高等学校化学学报, 1993, 14(9):1210-1213. [14]倪星忠, 舒为群. 丙酮酸氧化酶法测定ALT及其试剂盒的配制[J]. 检验医学, 1996, 11(2):75-77.
3. 毕业设计(论文)进程安排
2017.11-2017.12 文献查阅,完成开题报告,熟悉实验仪器 2018.1-2018.2 熟悉分子操作 2018.2-2018.3 质粒和菌株构建 2018.3-2018.4 丙酮酸氧化酶酶活的测定和酶的纯化 2018.4-2018.5 丙酮酸氧化酶在电极上的初步应用和论文撰写