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。

依靠本实验室筛选得到的微生物来源糖基转移酶催化人参皂苷，进行功能验证和产物鉴定，并通过改变反应条件，定向合成稀有人参皂苷。

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] 杨金玲, 高丽丽, 朱平. 人参皂苷生物合成研究进展[J]. 药学学报, 2013(2):170-178. [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] 梁会超, 王庆华, 巩婷, et al. 人参皂苷生物合成相关糖基转移酶研究基本策略及进展[J]. 药学学报, 2015(2):148-153. [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] 张南生, 张秀华, 李文峰. 人参皂苷Rg3的研究进展[J]. 医药导报, 2006, 25(7):687-689. [7] Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene fromBacillus subtilis: synthesis, characterization, andin vitromelanogenesis inhibition activity in BL6B16 cells[J]. Journal of Ginseng Research, 2018, 42(1):42-49. [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 2018.12.26-2019.1.16 文献查阅，完成开题报告，熟悉仪器 2019.3.1--2019.3.30 糖基转移酶的功能验证 2019.4.1--2019.4.30 稀有人参皂苷产物的鉴定 2019.5.5---2019.5.29 反应条件优化 2019.5.30---2019.6.6 撰写毕业论文