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摘要：甾体类药物近年来被广泛的应用于治疗各种疾病。雄甾-4-烯-3,17-二酮（AD）和雄甾-1,4-二烯-3,17-二酮（ADD）是甾体激素类药物的重要中间体，ADD几乎是所有甾体类激素药物生产的原料。随着薯蓣皂素等天然资源的日益枯竭，以廉价的植物甾醇制备ADD如今愈发受到重视。本实验以实验室储藏的一株分枝杆菌为出发菌株，以植物甾醇为底物，对发酵生产ADD的工艺流程进行了初步探索从而得出最优发酵参数。38699
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主要研究内容包括：对培养时pH的不同；接种菌液量的不同；加入不同的表面活性剂；加入不同的有机溶剂等因素进行了探究。又通过正交实验对接种菌液量，pH值，碳源浓度，诱导物浓度进一步优化。
初步确定最适接种菌液量为12%；最适pH为7.0；最适表面活性剂是0.2g/L DMF；最适有机溶剂为2.0% 的乙醇。微生物转化时间为8天。
毕业论文关键词： 植物甾醇，分枝杆菌，雄甾二烯二酮，生物转化
Microbial transformation of phytosterol production of pharmaceutical raw materials to the ADD process research
Abstract: Steroid drugs have been widely used in the treatment of various diseases in recent years. Androst -4- ene -3,17- dione(AD) and androsta -1,4- diene -3,17- dione (ADD) are the important intermediate of steroid hormones. Almost all of the steroid hormone drugs can use ADD as the starting material for the production. With the gradual depletion of some natural resources ang diosgenin , using phytosterols, the cheap and new materials, to produce ADD is growing in importance. This study used one mycobacterium strain which stored in laboratory as the original strain and phytosterols as the substrate. And the fermentation productivity of ADD in different process conditions was firstly explored. To find the optimal fermentation parameters is the core.
The main contents of the research was as follows: the difference in the types of pH when producing, the difference in the amount of inoculum, the difference in the types of organic solvent, etc. And the amount of inoculum, the pH value, the concentration of carbon source, inducer concentration were optimized by orthogonal test.
Preliminary determination of the most suitable amount of inoculum was 12%. The type of pH is 7.0. The most suitable surface active agent was DMF, the concentration was 0.2 g/L. The most suitable organic solvent was ethanol, the amount was 2.0%. The microbial transformation time was 8 days.
KeyWords： phytosterol；mycobacterium；androstenedione；Biotransformation
目录
1前言 1
1.1甾体激素药物的介绍 1
1.1.1甾体激素药物概述 1
1.1.2甾体激素药物的应用现状和研究 1
1.2植物甾醇甾醇概述 2
1.2.1甾体化合物与植物甾醇 2
1.2.2植物甾醇的种类及来源 3
1.2.3植物甾醇的功能及应用领域 3
1.3雄甾二烯二酮（ADD）的研究及应用 4
1.3.1 ADD的结构及理化性质 4
1.3.2制备ADD的方法 5
1.4微生物法转化植物甾醇生成ADD 6
1.4.1 微生物转化法的概述 6
1.4.2优化微生物转化反应 7
1.5本课题的研究内容及意义 9
2材料和方法 10
2.1材料与试剂 10
2.1.1菌种 10
2.1.2主要试剂 10
2.1.3主要材料 11
2.1.4主要仪器和设备 11
2.1.5培养基 11
2.2实验方法 11
2.2.1 实验前期准备 11
2.2.2 培养条件 12