摘  要目前利用发酵工程菌培养重组大肠杆菌是生产肠激酶的主要方法。发酵培养条件对肠激酶产量的提高有重要的影响。本实验对生产所用的菌种进行一系列的形态鉴定和生理生化检测，验证菌种的可用性；然后对可用的菌种进行发酵培养，通过对硫酸铵浓度、发酵温度、接种量、补料时间、诱导点的优化，对整个发酵条件进行优化，为增加肠激酶产量提供一些数据和方法。本实验得出的最优硫酸铵浓度是0.5%;最优接种量是5%；最优发酵温度是37℃；最佳补料时间是8h；最适诱导点是第14小时开始诱导。在此条件下进行发酵培养，可以使大肠杆菌的菌体浓度达到157g/L，包涵体的含量可以达到31g/L之高，很好的达到了优化的目的。69179

  该论文有图15幅，表8个，参考文献14篇。

毕业论文关键词：重组大肠杆菌  发酵培养  肠激酶    

Optimization of Production Conditions of Recombinant Intestinal Kinase

Abstract

At present, the main method of producing intestinal kinase by using recombinant Escherichia coli. The fermentation conditions had an important influence on the increase of the output of the intestinal kinase. In this experiment, the production of the strains were a series of morphological and physiological and biochemical tests, availability of validation strains; then the available strain fermentation, through the optimization of ammonium sulfate concentration optimization, inoculation amount, the optimization of fermentation temperature, fill time optimization, induced point of optimization, to optimize the fermentation process, provide some data and methods to increase the yield of enterokinase.Finally, the optimal concentration of ammonium sulfate is 0.5%, the optimal inoculation amount is 5%, the optimal fermentation temperature is 37 degrees C, the optimal feeding time is 8h; the optimal inducing point is fourteenth hours to start induction. Under the conditions of fermentation, the content of the inclusion body can reach 157g/L, and the content of the inclusion body can reach 31g/L, which is good to achieve the purpose of optimization.

Key words：recombinant E. coli  Fermentation culture  Intestinal kinas 

目   录

摘要 Ⅰ 

Abstract Ⅱ

目录 Ⅲ

图清单 Ⅳ

表清单 Ⅳ

1 绪论 5

1.1重组大肠杆菌生产肠激酶发展现状 5

1.2 研究目的及主要内容 5

2 材料和方法 6

2.1 试剂 6

2.2 仪器 6

2.3 实验菌种 7

2.4实验方法 7

3   结果与分析 11

3.1大肠杆菌菌种鉴定试验的结果和分析 11

3.2 重组大肠杆菌稳定性试验 15

3.3重组大肠杆菌生长曲线 15

3.4 重组大肠杆菌发酵过程条件优化 16

4 结论 24

参考文献 25

致谢 26 

图清单

图序号 图名称