双酶固定化生产高附加值糖基化产物任务书
2020-05-02 17:09:18
1. 毕业设计(论文)的内容和要求
论文内容: 我们依靠已有的糖基转移酶及蔗糖合成酶定向合成具有高附加值的稀有人参皂苷,并通过对双酶的协同固定,提升糖基化的稳定性及产率。
进一步的实验中,通过单因素法确定双酶固定的最优条件,使高附加值的糖基化产物产率达到最大,并使大批次合成成为可能。
论文要求: 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,查阅关于糖基转移酶研究现状,酶固定化方式优劣,多酶固定方式选择及条件优化方面的相关文献。
2. 参考文献
[1] 赵千婧,程瑶,王佳,孙新晓,申晓林,袁其朋.糖基转移酶合成相关糖苷类化合物研究进展[J].北京化工大学学报(自然科学版),2018,45(05):92-99. [2] 金月,吴旭日,陈依军.糖基转移酶在改善天然产物成药性方面的应用[J].中国药科大学学报,2017,48(05):529-535. [3] 金凤燮(主编).天然产物生物转化[M]. 北京:化学工业出版社,2009:74-101. [4] FEDOROFF N V,FURTEK D B,NELSON O E.Cloning of the bronze locus in maize by a simple and generalizable pocedure using the transposable controlling element activator (ac) [J]. Proc Nati Acad Sci USA,1984,81(12) : 3825-3829. [5] HUGHES J,HUGHES M A.Multiple secondary plant product UDP-glucose glucosyltransferase genes expressed in cassava (Manihot esculenta Crantz) cotyledons[J]. [6] SHAO H,HE X,ACHNINE L,et al. Crystal structures of a multifunctional triterpene /flavonoid glycosyltransferase from Medicago truncatula. [J]. Plant Cell,2005,17( 11) : 3141-3154. [7] HAYWARD A P,MORENO M A,HOWARDRT,et al.Control of sexuality by the skl-encoded UDP-glycosytransferase of Maize[J].Science Advances,2016,2(10) : e16009. [8] XIA T,EITEMAN M A. Quercetin glucoside production by engineered Escherichia coli[J].Appl Biochem Biotechnol,2017,182(4) : 1358-1370. [9] LUAN L C,PANDEY R P,LIM H N,et al. Synthesis of umbelliferone derivatives in Escherichia coli and their biological activities[J]. Journal of Biological Engineering,2017,11( 1) : 15. [10] PANDEY R P,PARAJULI P,GURUNG R B,et al. Donor specificity of Yjic glycosyltransferase determines the conjugation of cytosolic NDP-sugar in vivo glycosylation reactions[J]. Enzyme & Microbial Technology,2016,91: 26-33. [11] Gutmann A, Nidetzky B. Unlocking the Potential of Leloir Glycosyltransferases for Applied Biocatalysis: Efficient Synthesis of Uridine 5'-Diphosphate-Glucose by Sucrose Synthase[J]. Advanced Synthesis Catalysis, 2016, 358(22):3600-3609. [12] A.C.L.B. Gutmann, A two-step O- to C-glycosidic bond rearrangement using complementary glycosyltransferase activities, CHEMICAL COMMUNICATIONS, 50 (2014) 5465-5469. [13] B.C. Ahn, B.G. Kim, Y.M. Jeon, E.J. Lee, Y. Lim, J.-H. Ahn, Formation of flavone di-O-glucosides using a glycosyltransferase from Bacillus cereus, Journal of microbiology and biotechnology, 19 (2009) 387-390. [14] XU L J,QI T T,XU L,et al. Recent progress in the enzymatic glycosylation of phenolic compounds[J]. Journal of Carbo-hydrate Chemistry,2016,35(1) : 1-23. [15] C. Zhang, B.R. Griffith, Q. Fu, C. Albermann, X. Fu, I.-K. Lee, L. Li, J.S. Thorson, Exploiting the reversibility of natural product glycosyltransferase-catalyzed reactions, Science, 313 (2006) 1291-1294. [16] Wolf S , Berrio R M , Meier C . Synthesis of Nonnatural Nucleoside Diphosphate Sugars[J]. European Journal of Organic Chemistry, 2011, 2011(31):6304-6313. [17] Chem N , April P , Gantt R W , et al. Using Simple Donors to Drive the Equilibria of Glycosyltransferase-Catalyzed Reactions[J]. Nature Chemical Biology, 2011, 7(10):685. [18]梁华正,杨水平,丁志刚,饶军. 弱碱性大孔树脂固定化硫磺菌β- 葡萄糖苷酶的实验研究[J],食品开发与机械,No. 2. 2008,8-10 [19]陈秀琳等,CM-纤维素固定化脂肪酶的研究[J],海峡药学2005年第17卷第1期
3. 毕业设计(论文)进程安排
2018.12---期末 文献查阅,完成开题报告,熟悉仪器 2019.3---2019.4 了解实验用酶的基本功能、产物的检测及鉴定 2019.4---2019.6 对双酶进行固定,提升酶催化能力与稳定性 2019.5 优化固定化酶的条件 2019.5---2019.6 撰写毕业论文