1. 毕业设计（论文）的内容和要求

论文内容 本实验采用基因工程手段，挖掘糖基转移酶ugtsl2基因序列，并合理构建重组表达质粒。

以大肠杆菌为宿主诱导表达产酶，以莱胞迪甙a为底物，研究糖基转移酶ugtsl2催化合成莱胞迪甙d的性能，以及ugtsl2酶学性质的相关表征。

论文要求 1．文献查阅 掌握文献查阅的一般方法，学会利用互联网在中国期刊网、维普数据库、超星数字图书馆、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糖基转移酶基因及酶法合成莱胞迪甙d的相关文献，对本研究内容有广泛了解。

2. 参考文献

[1] Dewitte G., Walmagh M. (2016). Screening of recombinant glycosyltransferases reveals the broadacceptor specificity of stevia UGT-76G1. Journal of Biotechnology, 233, 49-55. [2] De Bruyn, F., Maertens, J., Beauprez, J., Soetaert, W., De Mey, M. (2015). Biotechnological advances in UDP-sugar based glycosylation of small molecules. Biotechnology Advances, 33(2), 288-302. [3] Georgiou, G., Valax, P. (1999). Isolating inclusion bodies from bacteria. Amyloid, Prions, And Other Protein Aggregates, 309, 48-58. [4] K#345;en, V., Thiem, J. (1998). Glycosylation employing bio-systems: from enzymes to whole cells. Cheminform, 29(14), no-no. [5] Lim, E. K. (2005). Plant glycosyltransferases: their potential as novel biocatalysts. Chemistry-a European Journal, 11(19), 5486-5494. [6] Madhav, H., Bhasker, S., Chinnamma, M. (2013). Functional and structural variation of uridine diphosphate glycosyltransferase (UGT) gene of Stevia rebaudiana-UGTSr involved in the synthesis of rebaudioside A. Plant Physiology And Biochemistry, 63(4), 245-253. [7] Olsson, K., Carlsen, S., Semmler, A., Simon, E., Mikkelsen, M. D., Moller, B. L. (2016). Microbial production of next-generation stevia sweeteners. Microbial Cell Factories, 15: 207. [8] Sauerzapfe, B., Engels, L., Elling, L. (2008). Broadening the biocatalytic properties of recombinant sucrose synthase 1 from potato (Solanum tuberosum L.) by expression in Escherichia coli and Saccharomyces cerevisiae. Enzyme And Microbial Technology, 43(3), 289-296. [9] Schmolzer, K., Gutmann, A., Diricks, M., Desmet, T., Nidetzky, B. (2016). Sucrose synthase: A unique glycosyltransferase for biocatalytic glycosylation process development. Biotechnology Advances, 34(2), 88-111. [10] Wang, J., Li, S., Xiong, Z., Wang, Y. (2016). Pathway mining-based integration of critical enzyme parts for de novo biosynthesis of steviolglycosides sweetener in Escherichia coli. Cell Res, 26(2), 258-261. [11] Wang, Y., Chen, L. L., Li, Y., Li, Y. Y., Yan, M., Chen, K. Q., Hao, N., Xu, L. (2016). Efficient enzymatic production of rebaudioside A from stevioside. Bioscience Biotechnology And Biochemistry, 80(1), 67-73.

3. 毕业设计（论文）进程安排

2018.1.11 -2018.2.24 查阅文献，熟悉课题有关背景，合理设计实验方案； 2018.2.25 -2018.3.25 将已有的含有糖基转移酶的重组质粒导入到大肠杆菌中，采用乳糖诱导培养基诱导产酶，催化莱胞迪甙A合成莱胞迪甙D； 2018.3.26 -2018.5.31 对粗酶液UGTSL2进行蛋白纯化，对酶的酶学性质进行进一步的研究； 2018.6.1 -2018.6.15 及时撰写毕业论文。