1. 毕业设计（论文）的内容和要求

在酶法合成嘌呤核苷类似物时，主要是用嘧啶核苷磷酸化酶水解嘧啶类核苷提供核糖供体，再加入嘌呤碱基，在嘌呤核苷磷酸酶的作用下催化合成嘌呤核苷。

其原因如下：1、嘧啶和嘌呤核苷磷酸化酶在热动力学方面分别偏向于水解合成，双酶催化有利于产物的累积。

2、第一步反应所产生的嘧啶碱基，不会参与第二步反应，不会因为积累而抑制第二步反应。

2. 参考文献

[1] Robak, T. and P. Robak, Purine nucleoside analogs in the treatment of rarer chronic lymphoid leukemias. Current Pharmaceutical Design, 2012. 18(23): p. 3373-3388. [2] Herschel Wallen, J.A.T.J., Fludarabine Modulates Immune Response and Extends In Vivo Survival of Adoptively Transferred CD8 T Cells in Patients with Metastatic Melanoma. Plos One, 2009. 4(3): p. e4749. [3] Bastin-Coyette, L., et al., Mechanisms of cell death induced by 2-chloroadenosine in leukemic B-cells. Biochemical Pharmacology, 2008. 75(75): p. 1451-1460. [4] Montgomery, J. and K. Hewson, Nucleosides of 2-Fluoroadenine. Journal of Medicinal Chemistry, 1969. 12(12): p. 498-504. [5] Lapponi, M.J., et al., New developments in nucleoside analogues biosynthesis: A review. Journal of Molecular Catalysis B Enzymatic, 2016. 133: p. 218-233. [6] Shugar D. Purine nucleoside phosphorylases: properties, functions, and chemotherapeutic applications[J]. Cellular Molecular Biology Letters, 1999, 04. [7] Serra I, Daly S, Alcantara A R, et al. Redesigning the synthesis of Vidarabine via a multienzymatic reaction catalyzed by immobilized nucleoside phosphorylases[J]. Rsc Advances, 2015, 5(30):23569-23577. [8] Trinh R, Gurbaxani B, Morrison S L, et al. Optimization of codon pair use within the (GGGGS) linker sequence results in enhanced protein expression[J]. Molecular Immunology, 2004, 40(10):717-722. [9] Sivaraj Sivaramakrishnan, James A. Spudich. Systematic control of protein interaction using a modular ER/K α-helix linker[J]. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(51):20467-20472. [10] Pugmire M J, Ealick S E. Structural analyses reveal two distinct families of nucleoside phosphorylases[J]. Biochemical Journal, 2002, 361(Pt 1):1-25. [11] 高鑫. 多酶在胞内外的超分子自组装研究[D]. 华东理工大学, 2016. [12] Zhu S, Song D, Gong C, et al. Biosynthesis of nucleoside analogues via thermostable nucleoside phosphorylase[J]. Applied Microbiology Biotechnology, 2013, 97(15):6769-6778. [13] Zhou, Xinrui, Szeker, Kathleen, Jiao, Lin‐Yu, et al. Synthesis of 2,6‐Dihalogenated Purine Nucleosides by Thermostable Nucleoside Phosphorylases[J]. Advanced Synthesis Catalysis, 2015, 357(6):1237-1244.

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 查阅、收集文献资料 2019.3.1～2019.3.30 融合蛋白linker的选择，以及引物的设计 2019.4.1～2019.4.30 融合蛋白表达菌株的构建以及表达 2019.5.1～2019.5.29 融合蛋白活性的测定及在合成中的应用 2019.5.30～2019.6.6 论文撰写及答辩