人参皂苷合成的酶基因分析与生物定向合成任务书
2020-06-25 20:51:17
1. 毕业设计(论文)的内容和要求
我们选择使用生物酶法定向合成所需的稀有人参皂苷。
依靠已有的糖基转移酶,测试其对人参皂苷的催化功能,并找出人参皂苷的催化位点与糖的连接情况。
进一步的实验中,一方面,对酶进行定点改造,优化催化功能和位点选择性;另一方面,测试催化的最适条件,摸索出大批次催化的途径。
2. 参考文献
[1] 王红艳,徐绥绪,陈英杰,等.人参单体成分药理活性研究的新进展, 中国药物化学杂志, 1992 , 2(3):73-78. [2]窦德强, 靳玲, 陈英杰. 人参的化学成分及药理活性的研究进展与展望.沈阳药科大学学报.1999,16(2):151-156. [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]TAWAB M A, BAHR U, KARAS M, et al. Degradation of ginsenosides in humans after oral administration [J]. Drug Metabolism Disposition, 2003, 31:1065-1071. [5] CAI B X, Luo D, Lin X F, et al. Compound K suppresses ultraviolet radiation-induced apoptosis by inducing DNA repair in human keratinocytes [J]. Archives of PharmacalResearch, 2008, 31(11):1483-1488. [6] ZHANG L, FU F, WANG T, et al. Effect of Compound K on chronic hepatic injury induced by carbon tetrachloride (C Cl4) in rats [J]. Ginseng Research, 2005, 28(6): 685-690. [7] SHIN Y W, KIM D H. Antipruritic effect of ginsenoside Rb1 and compound K in scratching behavior mouse models [J]. Journal of Pharmaceutical Sciences, 2005, 99(1): 83-88. [8] MATSUI T, KOHNO H, OTA T. Reduced sialylation of peanut agglutinin-binding sugar chains on CD44 associated with enhanced adhesiveness to endothelial cells and experimental metastatic ability in B16BL6 melanoma cells [J ]. Kanazawa Ika Daigaku Zasshi 1995, 20:194-204. [9] CHOI S G, KIM T W, SINGH S V. Ginsenoside Rh2-mediated G1 Phase Cell Cycle Arrest in Human Breast Cancer Cells Is Caused by p15Ink4B and p27Kip1-dependent Inhibition of Cyclin-dependent Kinases [J ]. Pharmaceutical Research, 2009, 26:2280-2288. [10] LIU T G, HUANG Y C, DAN D, et al. Inhibitory effect of ginsenoside Rg3 combined with gemcitabine on angiogenesis and growth of lung cancer in mice [J]. BMC Cancer,2009, 9:250-261. [11] LEE W H, CHOI J S, KIM H Y, et al. Heat-processed neoginseng, KG-135, down-regulates G1 cyclin-dependent kinase through the proteasome-mediated pathway in hela cells [J]. Oncology Reports, 2009, 21(2):467-474. [12] KIM S I, PARK J D, LEE Y H, et al. Preparation of 20(R)- and 20(S)-ginsenoside Rh1 from ginsenoside Re [J]. Koryo Insam Hakhoechi, 1991, 15(3):188-91. [13]刘惟瑳,陈英杰,刘明生等.人参皂苷Rh2的半合成[J]. 沈阳药学院学报,1988,5(1):14. [14] KIM B G, CHOI Y, KIM M R. Changes of ginsenosides in Korean red ginseng (Panax ginseng) fermented by Lactobacillus plantarum M1 [J]. Process Biochemistry, 2010,45:1319-1324. [15] YE L, ZHOU C Q, ZHOU W, et al. Biotransformation of ginsenoside Rb1 to ginsenoside Rd by highly substrate-tolerant Paecilomyces bainier 229-7 [J]. Bioresource Technology,2010, 101: 7872-7876. [16] HASEGAWA H, SUNG J H, MATSUMIYA S, et al. Main ginseng saponin metabolites formed by intestinal bacteria [J]. Planta Medica, 1996, 62(5):453-457. [17] BAE A, SHIN J, KIM D H. Metabolism of ginsenoside Re by human intestinal microflora and its estrogenic effect [J]. Biological Pharmaceutical Bulletin, 2005, 28(10):1903-1908. [18] SHIN H Y, LEE J H, LEE J Y, et al. Purification and characterization of ginsenoside Ra-hydrolyzing beta-D-xylosidase from Bifidobacterium breve K-110, a human intestinal anaerobic bacterium [J]. Biological Pharmaceutical Bulletin, 2003, 26:1170-1173. [19]DongAling, CuiYajun, CuoHonzhu et al. Microbiological Transformation of Ginsenoside Rgl〔J〕 药学学报2001,10(3):114-118. [20]赵立亚,鱼紅闪,金凤燮等.酶法生产稀有人参皂苷及其产物成分的分析[J].大连轻工学院学报,2002,21(2):112-115. [21]Qin Yan. Purification Method Improvement and Characterization of Novel Ginsenoside -Hydrolyzing β-Glucosidase from Paecilomyces Bainier sp.29 [J] . Biosci. 2008,72(2),352-359. [22]Ruan C C,Zhang H,Zhang L X,et al. Biotransformation of ginsenoside Rf to Rh1 by recombinant β-glucosidase[J]. Molecules,2009,14( 6) : 2043. [23]Kim J K,Cui C H,Yoon M H,et al. Bioconversion of major ginsenosides Rg1 to minor ginsenoside F1 using novel recombinant ginsenoside hydrolyzing glycosidase cloned from Sanguibacter keddieii and enzyme characterization[J]. J Biotechnol,2012,161( 3) : 294. [24]Hong H,Cui C H,Kim J K,et al. Enzymatic biotransformation of ginsenoside Rb1 and gypenoside XVⅡinto ginsenosides Rd and F2 by recombinant β-glucosidase from Flavobacterium johnson-iae [J]. J Ginseng Res,2012,36( 4) : 418. [25] Morozova O, Hirst M, Marra MA. Applications of new sequencing technologies for transcriptome analysis [J]. Annu Rev Genomics Hum Genet, 2009, 10: 135#8722;151. [26] Parkinson J, Blaxter M. Expressed sequence tags: an overview [J]. Methods Mol Biol, 2009, 533: 1#8722;12. [27] Ross J, Li Y, Lim E, et al. Higher plant glycosyltransferases [J]. Genome Biol, 2001, 2: REVIEWS3004. [28] Paquette S, Moller BL, Bak S. On the origin of family plant glycosyltransferases [J]. Phytochemistry, 2003, 62: 399#8722;413.
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 2017.12---期末 文献查阅,完成开题报告,熟悉仪器 2018.3---2018.4 了解实验用酶的基本功能、产物的检测及鉴定 2018.4---2018.6 对酶进行定点改造,提高催化能力和位点选择性 2018.5---2018.6 优化反应的条件 2018.6.2---2018.6.10 撰写毕业论文
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