[1] Sakaguchi K, Murao S. A new enzyme,penicillin-amidase [J]. J Agril Chem Soc Japan, 1950, 23: 411-416.

[2] Rolisonm G N. Formation of 6-aminopenicillin acid fron penicillin by enzymatic hydrolysis [J]. Nature, 1960, 187: 236-237.

[3] Vadamme E J. Enzyme involved in β-lactam antibiotic biosynthesis [J]. Adv Appl Microbiol, 1977, 21: 89-123.

[4] Vadamme E J, Votes J P. Microbial penicillin acylase [J]. Adv Appl Microbiol, 1974, 17: 311-369.

[5] Martin L, M.A.Prieto, E.Cortes, et al. Cloning and sequencing of the pac gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 [J].FEMS Microbiol Lett, 1995, 125: 2879-2923.

[6] Ohashi H, Katsuta Y, Hashizaume T, et al. Molecular cloning of the penicillin G acylase gene from Arthrobater viscosus [J]. Appl Environ Microbiol, 1988, 54(11):2 603-26063.

[7] Schumacher G, D Sizmann,H Haug, P Buckel, et al. Penicillin acylase from E.coli:unique gene-protein relation [J]. Nucleic Acids Res, 1986, 14: 5713-5727.

[8] Verhaert R M, Riemens A M, van der Laan J M, et al. Molecular cloning and analysis of the gene encoding the thermostable penicillin G acylase from Alcaligenes faecalis [J].Appl Microbiol, 2003, 62: 507-516.

[9] Plhackova K, Becka S, Skrob F, et al. Isolation and chataterization of a new strain of Achromobacter sp. with β-lactam antibiotic acylase activity [J]. Appl Environ Microbiol, 1997, 63(9): 3412-3418.

[10] Barbero J L, Buesa J M, de Buitrago GG, et al. Complete nucleotide sequence of the penicillin acylase gene from Kluyvera citrophila [J]. Gene, 1986, 49: 69-80.

[11] Ljubijankic G M, Konstantinovic, and Glisin V. The primary structure of Providencia rettgeri penicillin G amidase gene and its relationship to other gram negative amidases [J]. DNA Seq, 1992, 3: 195-200.

[12] A Matsuda, K I Komatsu. Molecular cloning and structure of the gene for 7-beta-(4-carboxybutanamido) cephalosporanic acid acylase from a Pseudomonas strain [J]. J.Bacteriol, 1985, 163(3): 1222-1228.

[13] Be#269;ka S, #352;těp#225;nek V, Vyasarayani R W, et al. Penicillin G acylase from Achromobacter sp. CCM 4824[J]. Applied microbiology and biotechnology, 2014, 98(3): 1195-1203.

2015.12～2016.1

查阅、收集文献资料

阅读文献综述、开题报告。

2016.3～2016.3

理论学习及总体方案规划

了解实验的目的，规划最佳实验方案。

2016.3～2016.4

青霉素酰化酶连接6个组氨酸标签构建

构建青霉素酰化酶基因重组质粒，导入大肠杆菌宿主内高效表达。

2016.4～2016.4

发酵

发酵通过破碎得到青霉素酰化酶。

2016.4～2016.5

青霉素酰化酶的蛋白纯化

摸索出Ni纯化的最佳条件，对青霉素酰化酶进行蛋白纯化。

2016.5～2016.5

论文撰写

完成论文。