摘要本文探究了纯菌株催化精氨酸脱亚胺产生瓜氨酸的工程菌的培养方法和固定化方法，同时利用硅胶薄层层析的方法检测瓜氨酸的生成和精氨酸的消耗。工程菌经过筛选和诱导培养后具有精氨酸脱亚胺活性，用海藻酸钠和聚乙烯醇固定化后的工程菌实现了催化精氨酸脱亚胺生成瓜氨酸。本实验中工程菌产生的产精氨酸脱亚胺酶是一种胞内酶，工程菌大量表达后形成包涵体，使得提取细胞内产精氨酸脱亚胺酶催化精氨酸脱亚胺效率较低，所以通过固定化工程菌的方法实现高效催化精氨酸脱亚胺并且回收固定化工程菌到达提高工程菌利用率的方法有较好的利用前景。同时，精氨酸和瓜氨酸的产品价格悬殊，催化生成的瓜氨酸在医疗，药物前体等领域的市场广大，所以产精氨酸脱亚胺酶工程菌的固定化开发也有较大的应用前景和经济价值。18901
关键字：工程菌 固定化 产精氨酸脱亚胺酶 精氨酸 瓜氨酸
毕业设计说明书（毕业论文）外文摘要
Title Production engineering arginine deiminase immobilized bacteria
Abstract
This paper explores the catalytic arginine deiminase pure strain culture method citrulline engineering bacteria and immobilization methods produce while using silica gel thin layer chromatography method for detecting the generation of citrulline and arginine consumption. Engineered bacteria after screening and induction training with arginine deiminase activity, polyvinyl alcohol, sodium alginate and immobilized engineering bacteria to achieve a catalytic arginine deiminase generate citrulline. The experiment project arginine producing bacteria produce deiminase is an intracellular enzyme, the formation of large inclusion bodies after expression of engineering bacteria, making the extraction of cells producing arginine deiminase Arginine deiminase catalytic less efficient, so efficient catalytic method immobilized by arginine deiminase engineered bacteria immobilized and recovery methods to improve engineering bacteria engineered bacteria to reach a better utilization utilization prospects. Meanwhile, arginine and citrulline price disparity, the catalytic generation of citrulline in the field of medical, pharmaceutical precursors, such as the majority of the market, so the production of arginine deiminase immobilized engineered bacteria have developed a greater application prospects and economic value.
Keywords: engineering bacteria, immobilized, arginine deiminase produced citrulline, Arginine, citrulline
目 次
1 引言 1
1.1 精氨酸介绍 1
1.2 瓜氨酸概况 1
1.3 本次研究的意义 3
2 精氨酸脱亚胺酶 4
2.1 精氨酸脱亚胺酶活性的检测 4
2.2 产精氨酸脱亚胺酶工程菌活性的检测 5
3 实验材料和试剂 6
3.1 主要仪器设备 6
3.2 主要原料试剂 7
3.3 实验菌株 8
3.4 常用溶液 8
4 实验方法和结果表征 8
4.1 筛选培养基的制备 8
4.2 工程菌的活化 9
4.3 工程菌的种子培养 9
4.4 发酵培养和诱导 10
4.5 工程菌产精氨酸脱亚胺酶活性的检测 10
4.6 工程菌发酵生产瓜氨酸的探究 10
5 产精氨酸亚胺酶工程菌的固定化 14
5.1 菌株 14
5.2 主要材料和试剂 15
5.3 工程菌固定化方法 15 产精氨酸脱亚胺酶工程菌的固定化:http://www.youerw.com/shengwu/lunwen_40204.html