1. 毕业设计（论文）的内容和要求

论文内容 本论文包括对于纯酶pgapx02进行酶学性质研究，包括最适ph、ph稳定性、最适温度、温度稳定性以及有机溶剂耐受性。

并且研究不同的反应条件对其合成头孢羟氨苄的影响，包括ph，温度，底物摩尔比，加酶量等，确认最优的反应条件，得到产率。

论文要求 1．文献查阅 掌握文献查阅的一般方法，学会使用计算机在中国期刊网，维普数据库，超星数字图书馆，美国化学学会（acs）数据库，ca，sci等检索资源上查阅关于青霉素合成半合成类抗生素的相关文献。

2. 参考文献

[1] Illanes A, Altamirano C, Fuentes M, et al. Synthesis of cephalexin in organic medium at high substrate concentrations and low enzyme to substrate ratio[J]. Journal of Molecular Catalysis B: Enzymatic, 2005, 35(1): 45-51. [2] Maxin I Youshko, Luuk M van Langen, et al. Penicillin acylase-catalyzed synthesis of amipicillin in ”aqueous solution-precipitate”system.high substrate concentration and supersaturation effect [J]. J Mol Catal B:Enzym, 2000, 10: 509-515. [3] Terreni M, Tchamkam J, Sarnataro U, et al. Influence of Substrate Structure on PGA-Catalyzed Acylations. Evaluation of Different Approaches for the Enzymatic Synthesis of Cefonicid[J]. Advanced Synthesis Catalysis, 2005, 347(1):121#8211;128. [4] 孔佳佳. 青霉素酰化酶、其高产菌株及应用: CN 103184182 A [P]. 2013-04-11. [5] 施晓疌, 王永红, 张嗣良, 等. NIPAB法测定青霉素G酰化酶活力的分析和改进 [J]. 中国医药工业杂志, 2008, 39(9): 659-661. [6] Srirangan K, Orr V, Akawi L, et al. Biotechnological advances on penicillin G acylase: pharmaceutical implications, unique expression mechanism and production strategies[J]. Biotechnology advances, 2013, 31(8): 1319-1332. [7] Van Langen L M, Oosthoek N H P, et al. Penicillin acylase catalysed of ampicillin in hydrophilic organic solvents [J]. Biochem J, 1969, 115(4): 733-739 [8] Diender M B, Straathol A J J, et al. Course of pH during the formation of amoxicillin by suspension-to-suspesion reaction [J]. Enzyme Microb Technol, 2000, 27: 576-582. [9] Illanes A, Wilson L, Corrotea O, et al. Synthesis of cephalexin with immobilized penicillin acylase at very high substrate concentrations in fully aqueous medium [J]. J Mol Catal B:Enzym, 2007, 47(1-2): 72-78. [10] Goncalves L R B, Sousa R Jr, Fernandez-Lafuente R, et al.Enzymatic synthesis of amoxicillin avoiding limitations of the mechanistic approach for reaction kinetics [J]. Biotechnol Bioeng, 2002, 80(6): 622-631. [11] Fern#225;ndez-Lafuente R, Rosell C M, Piatkowska B, et al. Synthesis of antibiotics (cephaloglycin) catalyzed by penicillin G acylase: Evaluation and optimization of different synthetic approaches [J]. Enzyme Microbial Technology, 1996, 19(1):9-14. [12] Terreni M, Ubiali D, Bavaro T, et al. Enzymatic synthesis of cephalosporins. The immobilized acylase from Arthrobacter viscosus: a new useful biocatalyst. [J]. Applied Microbiology Biotechnology, 2008, 77(3):579-87. [13] Bazzini C, Brovelli S, Caronna T, et al. Characterization and study of the orientation of immobilized enzymes by tryptic digestion and HPLC-MS: design of an efficient catalyst for the synthesis of cephalosporins. [J]. Biomacromolecules, 2010, 11(6):1623-32. [14] Cecchini D A, Pavesi R, Sanna S, et al. Efficient biocatalyst for large-scale synthesis of cephalosporins, obtained by combining immobilization and site-directed mutagenesis of penicillin acylase.[J]. Applied Microbiology Biotechnology, 2012, 95(6):1491-500. [15] Franti#353;ek #352;krob, Stanislav Be#269;ka, Kamila Plh#225;#269;kov#225;, et al. Novel penicillin G acylase from Achromobacter, sp. CCM 4824[J]. Enzyme Microbial Technology, 2003, 32(6):738-744. [16] Be#269;ka S, #352;těp#225;nek V, Vyasarayani R W, et al. Penicillin G acylase from Achromobacter sp. CCM 4824: an efficient biocatalyst for syntheses of beta-lactam antibiotics under conditions employed in large-scale processes. [J]. Applied Microbiology Biotechnology, 2014, 98(3):1195-203.

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 2017.1-2017.2 查阅文献，了解课题内容 2017.2-2017.3 酶学性质测定 2017.3-2017.4 部分合成条件优化，pH、温度、加酶量 2017.4-2017.5 其余合成条件优化，底物摩尔比，有机溶剂等 2017.5-2017.6 论文攥写