1. 毕业设计（论文）的内容和要求

论文内容： 稀有人参皂苷是人参中的主要功能成分，但不同皂苷单体在药材及其提取物中量不同，其药理活性亦有差异，量低而药理活性强的皂苷单体需大量制备。

如何定向提高人参皂苷单体的量，酶法制备是解决该问题的关键技术。

依靠已报道糖基转移酶作为模板，寻找并筛选出微生物来源的糖基转移酶，克隆表达，并验证功能。

2. 参考文献

[1] Tita Ritsema, Jeanine Joling Sjef Smeekens. Patterns of fructan synthesized by onion fructan:fructan 6G-fructosyltransferase expressed in tobacco BY2 cells is fructan: fructan 1-fructosyltransferase needed in onion [J].New Phytologist, 2003, 160: 61-67 [2] 李玉 , 路福平 , 王正祥. 功能性低聚糖合成中糖基转移酶研究进展 [J]. 食品科学,2013,34(9):358-363 [3] Jia L, Zhao Y. Current evaluation of the millennium phytomedicine-ginseng (I): Etymology, pharmaco-gnosy, phytochemistry, market and regulations [J]. Curr Med Chem, 2009, 16(19): 2475-2484. [4] 毛多斌,王雪.果糖基转移酶及低聚果糖生产研究进展[J].广东化工,2009,4(36):100-102 [5] 陈利平,韩亚伟,毛多斌,等.枯草芽胞杆菌果糖基转移酶基因的克隆及其序列分析[J].河南农业大学学报,2009,4(43):432-436 [6] 唐博,崔子寅,王艺萌,等.在大肠杆菌中高效表达具有抗病毒活性的可溶性犬α7 干扰素[J].中国预防兽医学报,2013,4(35):321-324 [7] 何小妮,蒋波,王玉海,等.非对称灭活双亲原生质体融合法选育产果糖基转移酶的米曲霉新菌株的研究[J].现代食品科技,2013,29(5):993-997 [8] Quan L H, Min J W, Jin Y, et al. Enzymatic biotransformation of ginsenoside Rb1 to 20 (S)-Rg3 by recombinant β-glucosidase from Microbacterium esteraromaticum [J]. Appl Microbiol Biotechnol, 2012, 94(2): 377-384. [9] Keegstra K, Raikhel N. Plant glycosyltransferases. Current opinion in plant biology, 2001,4(3):219~224.6 Coutinho PM, Deleury E, Davies GJ, et al. An evolving hierarchical family classification for glycosyltransferases. Journal of molecular biology,2003, 328(2):307~317. [10] Zhao L G, Xie J C, Zhang X S, et al. Over expression and characterization of a glucose-tolerant-glucosidase from Thermotoga thermarum DSM 5069T with high catalytic efficiency of ginsenoside Rb1 to Rd [J]. J Mole Cataly B: Enzym, 2013, 95: 62-69. [11] Zhao X S, Gao L, Wang J, et al. A novel ginsenoside Rb1-hydrolyzing β-D-glucosidase from Cladosporium fulvum [J]. Process Biochem, 2009, 44(6): 612-618. [12] Fukuchi-Mizutani M, Okuhara H, Fukui Y, et al. Biochemical and molecular characterization of a novel UDP-glucose: anthocyanin 3'-O-glucosyltransferase, a key enzyme for blue anthocyanin biosynthesis, from gentian. Plant physiology, 2003, 132(3):1652. [13] Jones P, Vogt T. Glycosyltransferases in secondary plant metabolism: tranquilizers and stimulant controllers. Planta, 2001, 213(2):164~174. [14] Poppenberger B, Berthiller F, Lucyshyn D, et al. Detoxification of the Fusarium mycotoxin deoxynivalenol by a UDP #8211;glucosyltransferase from Arabidopsis thaliana. Journal of Biological Chemistry,2003, 278(48):47905~47914. [15] Zagrobelny M, Bak S, Rasmussen AV, et al. Cyanogenic glucosides and plant-insect interactions. Phytochemistry, 2004, 65(3):293~306. [16] Cheng L Q, Kim M K, Lee J W, et al. Conversion of major ginsensodie Rb1 to ginsenoside F2 by Caulobacter leidyia [J]. Biotechol Lett, 2006, 28(14): 1121-1127. [17] Staswick PE, Tiryaki I. The oxylipin signal jasmonic acid is activated by an enzyme that conjugates it to isoleucine in Arabidopsis. The Plant Cell, 2004, 16(8):2117. [18] Bowles D, Isayenkova J, Lim EK, et al. Glycosyltransferases: managers of small molecules.Current opinion in plant biology,2005, 8(3):254~263. [19] Liu L, Zhu X M, Wang Q J, et al. Enzymatic preparation of 20 (S, R)-protopanaxadiol by transformation of 20 (S,R)-Rg3 from black ginseng [J]. Phytochemistry, 2010, 71(13): 1514-1520. [20] Quan L H, Wang C, Yang J, et al. Isolation and characterization of novel ginsenoside-hydrolyzing glycosidase from Microbacterium esteraromaticum that transforms ginsenoside Rb2 to rare ginsenoside 20 (S)-Rg3[J]. Antonie Leeuwenhoek, 2013, 104(1): 129-137.

3. 毕业设计（论文）进程安排

起讫日期 设计（论文）各阶段工作内容 备 注 2016.12---期末 文献查阅，完成开题报告，熟悉仪器 2017.3---2017.4 确定克隆表达的酶及来源菌株 2017.4---2017.5 进行酶的克隆表达及优化发酵 2017.5---2017.6 确定找到的糖基转移酶的功能 2017.6.2---2017.6.10 撰写毕业论文