海狸磁珠固定化腈水解酶及其催化水解1-氰基-环己基乙腈的研究
毕业论文关键词:腈水解酶;固定化; 磁性材料
Studies on nitrilase immobilized by beaver magnetic particles and catalyze hydrolyze 1-Cyanocyclohexaneacetonitrile
Abstract：Nitrilase is such an enzyme that can convert nitriles into carboxylic acid or ammonium. Compared with the harsh condition of chemical synthesis, such as high pressure, high temperature, poisonous heavy metal catalysis and alkali, catalysis through enzyme brings about high value and wide application under mild condition in organic synthesis and industrial production. Magnetic immobilized enzyme not only help to improve the stability (operation stability and convenience , storage stability , heat stability) , but also it is conducive to the subsequent separation of the reaction product of a continuous process , etc., and the reuse of the enzyme is to solve waste. After systematic optimization, the best reaction condition is the following: Buffer A, immobilization time was 30 min, Amount of beaver magnetic particles is 3 ml, nitrilase concentration is 6 g/L ,the optimum pH value of 7.0.
Key words: nitrilase; immobilization; magnetic materials
目 录
1. 研究背景 1
1.1 腈水解酶简介 1
1.2 生物催化 1
1.3固定化 1
1.4 加巴喷丁 2
1.5酶固定化的方法 2
1.6腈水解酶的应用 3
1.7磁性高分子微球的特点 3
1.8磁性微球固定化研究现状 3
1.9 研究的意义和内容 4
1.9.1研究意义 4
1.9.2研究内容 4
2. 实验原理和流程 5
3. 试验方法与材料 6
3.1 实验材料 6
3.1.1 菌种 6
3.1.2 培养基 6
3.1.3 自配试剂 6
3.1.4 主要试剂 6
3.1.5 主要仪器 7
3.2 实验方法 8
3.2.1 分析方法 8
3.2.2 菌体培养 8
3.2.3 测定游离细胞的酶活 8
3.2.4 大肠杆菌的破碎 9
3.2.5腈水解酶的固定化 9
3.2.6 磷酸盐缓冲液与BufferA缓冲液固定化腈水解酶的研究 9
3.2.7固定化时间的优化 9
3.2.8 固定化中磁珠量的优化 10
3.2.9粗酶浓度的研究 10
3.2.10 固定化酶最适PH的研究 10
3.2.11 固定化酶pH稳定性的研究 10
4. 实验结果和讨论 11
4.1固定化缓冲液的优化 11
4.2固定化时间的优化 13
4.3磁珠量的研究 14
4.4粗酶浓度的研究 15
4.5最适pH研究 16
4.6 pH稳定性研究 17
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