1. 毕业设计（论文）的内容和要求

1．文献查阅 掌握文献查阅， 使用网络资源如中国期刊网、 维普数据库、 超星数字图书 馆、 elsevier 电子期刊、 springer link 全文电子期刊等检索资源， 查阅关于大丙酮酸氧化酶生物传感器方面的相关文献。

2． 文献阅读及综述 阅读与课题相关的外文与中文文献，了解国内外的研究动态， 撰写文献综 述。

3． 明确实验任务， 拟定实验方案 根据所查阅文献的内容， 确定实验内容及方案， 拟定科学可行的研究计划。

2. 参考文献

[1]Zapata-Bacri A M , Burstein C . Enzyme electrode composed of the pyruvate oxidase from pediococcus species coupled to an oxygen electrode for measurements of pyruvate in biological media[J]. Biosensors, 1987, 3(4):227-237.. [2]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. [3]Risse B , G#195;#188;nter Stempfer, Rudolph R , et al. Characterization of the stabilizing effect of point mutations of pyruvate oxidase from lactobacillus plantarum: Protection of the native state by modulating coenzyme binding and subunit interaction[J]. Protein Science, 1992, 1(12):1710-1718. [4]Gajovic N , Binyamin G , Warsinke A , et al. Operation of a Miniature Redox Hydrogel-Based Pyruvate Sensor in Undiluted Deoxygenated Calf Serum[J]. Analytical Chemistry, 2000, 72(13):2963-2968. [5]Gajovic N , Katja Haberm#252;ller, Warsinke A , et al. A Pyruvate Oxidase Electrode Based on an Electrochemically Deposited Redox Polymer[J]. Electroanalysis, 2015, 11(18):1377-1383. [6]Gavalas V G , Chaniotakis N A . Phosphate biosensor based on polyelectrolyte-stabilized pyruvate oxidase[J]. Analytica Chimica Acta, 2001, 427(2):271-277. [7]Sedewitz B , Schleifer K H , F G#246;tz. Physiological role of pyruvate oxidase in the aerobic metabolism of Lactobacillus plantarum.[J]. Journal of Bacteriology, 1984, 160(1):462-5. [8]Juan E C M , Hoque M M , Hossain M T , et al. The structures of pyruvate oxidase from Aerococcus viridans with cofactors and with a reaction intermediate reveal the flexibility of the active-site tunnel for catalysis[J]. Acta Crystallographica Section F Structural Biology and Crystallization Communications, 2007, 63(Pt 11):900-907. [9]Ikebukuro K , Wakamura H , Karube I , et al. Phosphate sensing system using pyruvate oxidase and chemiluminescence detection[J]. Biosensors Bioelectronics, 1996, 11(10):959-965. [10]Kulys J , Wang L , Daugvilaite N . Amperometric methylene green-mediated pyruvate electrode based on pyruvate oxidase entrapped in carbon paste[J]. Analytica Chimica Acta, 1992, 265(1):15-20. [11]Kwan R C H , Leung H F , Hon P Y T , et al. A screen-printed biosensor using pyruvate oxidase for rapid determination of phosphate in synthetic wastewater[J]. Applied Microbiology and Biotechnology, 2005, 66(4):377-383. [12]Mascini M , Mazzei F . Amperometric sensor for pyruvate with immobilized pyruvate oxidase[J]. Analytica Chimica Acta, 1987, 192(1):9-16. [13]Muller Y , Schulz G . Structure of the thiamine- and flavin-dependent enzyme pyruvate oxidase[J]. Science, 1993, 259(5097):965-967. [14]Muller Y A , G#252;nter Schumacher, Rudolph R , et al. The Refined Structures of a Stabilized Mutant and of Wild-type Pyruvate Oxidase from Lactobacillus plantarum[J]. Journal of Molecular Biology, 1994, 237(3):0-335. [15]Rahman M A , Park D S , Chang S C , et al. The biosensor based on the pyruvate oxidase modified conducting polymer for phosphate ions determinations[J]. Biosensors Bioelectronics, 2006, 21(7):1116-1124. [16]Zhang W , Chang H , Rechnitz G A . Dual-enzyme fiber optic biosensor for pyruvate[J]. Analytica Chimica Acta, 1997, 350(1-2):59-65.

3. 毕业设计（论文）进程安排

2018.12-2019.02 文献查阅， 完成开题报告， 熟悉实验仪器 2019.03-2019.04 通过蛋白纯化得到丙酮酸氧化酶纯酶液 2019.04-2019.05 选用戊二醛等固定化材料对丙酮酸氧化酶的固定化效果进行初步考察并计算酶活回收率， 2019.05-2019.06 优化固定化体系，进行固定化酶电极表征， 整理实验数据，撰写毕业论文