1. 毕业设计（论文）的内容和要求

论文内容 本研究以大肠杆菌为研究对象，采用分子克隆技术，构建糖基转移酶ugt88a1与蔗糖合酶sus共表达重组质粒，采用iptg诱导培养基诱导表达产酶，利用全细胞催化手段催化槲皮素合成槲皮素-3-o-葡萄糖糖苷和槲皮素-4'-o-葡萄糖糖苷。

同时，在全细胞催化反应中，研究各条件因素对双酶偶联反应体系中底物转化率的影响，以及对目标产物进行定量分析。

论文要求 （1）掌握文献查阅的一般方法，学会在中国期刊网、web of science科学引文索引、springer link电子期刊、elsevier sdos电子期刊等检索资源上查阅关于生物催化槲皮素研究的相关文献，并对大肠杆菌表达系统有全面了解。

2. 参考文献

[1] Eng-Kiat Lim D A A B, Rosamond G. Jackson D J B. Arabidopsis Glycosyltransferases as Biocatalysts in Fermentation for Regioselective Synthesis of Diverse Quercetin Glucosides [J]. BIOTECHNOLOGY AND BIOENGINEERING, 2004, 87(5): 623~631. [2] Ren G, Hou J, Fang Q, et al. Synthesis of flavonol 3-O-glycoside by UGT78D1 [J]. Glycoconjugate Journal, 2012, 29(5-6): 425~432. [3] Johnson MH D M E F. Anthocyanins and proanthocyanidins from blueberry-blackberry fermented beverages inhibit markers of in#64258;ammation in macrophages and carbohydrate-utilizing enzymes in vitro [J]. Mol Nutr Food Res, 2013, 57: 1182~1197. [4] Ahn J. Production of Flavonoid O-Glucoside Using Sucrose Synthase and Flavonoid O-Glucosyltransferase Fusion Protein [J]. Journal of Microbiology and Biotechnology, 2009. [5] Li L M L E L. Crystal structure of Medicago truncatula UGT85H2#8212;insights into the structural basis of a multifunctional (iso) #64258;avonoid glycosyltransferase [J]. J Mol Biol, 2007, 370: 951~963. [6] Jeon YM K B K J. Enzymatic glycosylation of phenolic compounds using BsGT-3 based on molecular docking simulation [J]. J Korean Soc Appl Biol Chem, 2009, 52: 98~101. [7] Gosch C H H S B. Cloning and heterologous expression of glycosyltransferases from Malus x domestica and Pyrus communis, which convert phloretin to phloretin 2′-O-glucoside (phloridzin) [J]. Plant Sci, 2010, 178: 299~306. [8] Yi Li, Sandie Baldauf, Eng-Kiat Lim, and Dianna J. Bowles. Phylogenetic Analysis of the UDP-glycosyltransferase Multigene Family of Arabidopsis thaliana. THE JOURNAL OF BIOLOGICAL CHEMISTRY. (2001), Vol. 276, No. 6, 4338#8211;4343. [9] Lim EK A D H B. Arabidopsis Glycosyltransferases as Biocatalysts in Fermentation for Regioselective Synthesis of Diverse Quercetin Glucosides [J]. Biotechnol Bioeng, 2004, 87(5): 623~631. [10] 冯锋,柳文媛,陈优生,刘静涵,赵守训.菝葜中黄酮和芪类成分的研究[J].中国药科大学学报,2003(02):23-25.

3. 毕业设计（论文）进程安排

2018.2.26-2018.3.2：做预实验，给后期实验提供指导意义； 2018.3.3-2018.3.9：查阅文献，并结合预实验结果分析，设计合理实验方案； 2018.3.10-2018.5.4：将已有的含有双酶的重组质粒导入到大肠杆菌中，采用IPTG诱导培养基诱导产酶；初步建立双酶催化槲皮素合成槲皮素-3-O-葡萄糖糖苷和槲皮素-4'-O-葡萄糖糖苷体系； 2018.5.5-2018.5.31：研究各条件因素对全细胞反应体系底物转化率的影响，并合理优化提高产物产率，同时及时整理、分析实验数据； 2018.6.1-2018.6.15：及时撰写毕业论文。