1. 毕业设计（论文）的内容和要求

核苷磷酸化酶广泛存在与动植物和微生物中，主要参与细胞核酸代谢途径中的代谢，它们在补救途径中催化核苷或脱氧核苷的糖苷键的可逆磷酸化反应。

在医药工业中主要用来合成抗病毒抗肿瘤的核苷类似物药物。

本论文主要通过核苷磷酸化酶作为生物催化剂来合成用于治疗急性白血病的6-巯基嘌呤核苷。

2. 参考文献

[1] 夏俊刚，何逵夫，谢希贤，等.枯草芽孢杆菌嘌呤核苷磷酸化酶酶学性质研究及在利巴韦林酶法合成中的应用[J].中国生物工程杂志，2010，30(12):53-59. [2] 吴晓，倪孟祥，朱晓丽,等.利用嘧啶核苷磷酸化酶基因工程菌酶法合成5-氟尿苷的转化率影响因素的研究[J].药物生物技术，2008，15(4):282-285. [3] Stepchenko, V.A., et al., Enzymatic Synthesis of 2-Deoxy-B-D-ribonucleosides of 8-Azapurines and 8-Aza-7-deazapurines. Synlett, 2012. 23(10): p. 1541. [4] Konstantinova, I.D., et al., [Biotechnological synthesis of ribavirin. Effect of ribavirin and its various combinations on the reproduction of Vaccinia virus]. Bioorg Khim, 2004. 30(6): p. 613-620. [5] Zhou X, Mikhailopulo I A, Bournazou M N C, et al. Immobilization of thermostable nucleoside phosphorylases on MagReSyn 庐;, epoxide microspheres and their application for the synthesis of 2,6-dihalogenated purine nucleosides[J]. Journal of Molecular Catalysis B Enzymatic, 2015, 115:119-127. [6] Zhou X, Szeker K, Jiao L, et al. Synthesis of 2,6‐Dihalogenated Purine Nucleosides by Thermostable Nucleoside Phosphorylases[J]. Advanced Synthesis amp; Catalysis, 2015, 357(6):1237-1244. [7] Iglesias L E, Lewkowicz E S, Medici R, et al. Biocatalytic approaches applied to the synthesis of nucleoside prodrugs.[J]. Biotechnology Advances, 2015, 33(5):412-434. [8] Bagarolo M L, Porcelli M, Martino E, et al. Multiple disulfide bridges modulate conformational stability and flexibility in hyperthermophilic archaeal purine nucleoside phosphorylase[J]. Biochimica et biophysica acta, 2015, 1854(10 Pt A):1458. [9] Montgomery, J.A. and K. Hewson, SYNTHESIS OF POTENTIAL ANTICANCER AGENTS. X. 2-FLUOROADENOSINE1. Journal of the American Chemical Society, 2002. 79(16): p. 4559. [10] Zhou, X., et al., Synthesis of 2,6-ihalogenated Purine Nucleosides by Thermostable Nucleoside Phosphorylases. Advanced Synthesis amp; Catalysis, 2015. 357(6): p. 1237-1244. [11] Taran, S.A., et al., Synthesis of 2-chloro-2-deoxyadenosine by microbiological transglycosylation using a recombinant Escherichia coli strain. Applied Biochemistry and Microbiology, 2008. 44(2): p. 162-166. [12] Ilja V Fateev, K.V.A.I., The chemoenzymatic synthesis of clofarabine and related 2 deoxyfluoroarabinosyl nucleosides: the electronic and stereochemical factors determining substrate recognition by E. coli nucleoside phosphorylases. Beilstein Journal of Organic Chemistry, 2014. 10(1): p. 1657. [13] Nbile, M., et al., Use of Citrobacter koseri whole cells for the production of arabinonucleosides: A larger scale approach. Process Biochemistry, 2012. 47(12): p. 2182-2188. [14] Fern#225;ndez-Lucas J, Fresco-Taboada A, De l M I, et al. One-step enzymatic synthesis of nucleosides from low water-soluble purine bases in non-conventional media.[J]. Bioresource Technology, 2012, 115(115):63-9. [15] Vande V J, Liekens S, Mcguigan C, et al. The cytostatic activity of NUC-3073, a phosphoramidate prodrug of 5-fluoro-2'-deoxyuridine, is independent of activation by thymidine kinase and insensitive to degradation by phosphorolytic enzymes.[J]. Biochemical Pharmacology, 2011, 82(5):441-52. [16] Zhu S, Song D, Gong C. Biosynthesis of nucleoside analogues via thermostable nucleoside phosphorylase[J]. Applied Microbiology and Biotechnology, 2013, 97(15):6769-78.

3. 毕业设计（论文）进程安排

2017.12～2018.1 查阅、收集文献资料 2018.3～2018.3 理论学习及总体方案规划 2018.3～2018.4 嘧啶核苷磷酸化酶和嘌呤核苷磷酸化酶最适反应温度，最适反应pH的测定 2018.4～2018.4 双酶温度稳定性，pH稳定性，有机溶剂耐受性的测定 2018.4～2018.5 6-巯基嘌呤核苷合成条件的优化 2018.5～2018.5 论文撰写