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摘要：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，几乎所有甾体激素药物都可以以ADD为起始原料进行生产。随着薯蓣皂素等天然资源的日渐枯竭，利用化学合成制备ADD的方法已不再适用。植物甾醇以其成本低廉、获取便利逐渐成为制备ADD的新原料。近年来，利用分枝杆菌降解植物甾醇制备ADD的研究已经广泛开展。本文以实验室保藏的一株分枝杆菌为出发菌株，以大豆甾醇为底物，对发酵生产ADD的工艺流程进行了初步探索，主要完成了以下几个方面的工作：对培养基的碳源、氮源、的选择以及铁离子和及其抑制剂进行了探究，对培养温度、底物加入量等因素进行了考察，然后利用正交试验对碳源浓度、氮源浓度、底物加入时间、诱导物浓度进行了优化。初步确定培养基最适碳源为葡萄糖，浓度为7.5g/L，最适氮源为磷酸二氢铵，浓度为2g/L，培养基中加入0.005g/L柠檬酸铁铵，为菌体生长补充铁离子，同时在底物加入前加入0.025g/L751水合氯化钴，最适培养温度为30℃，最佳底物加入量为2g/L，最适底物加入时间为24h，采用AD作为诱导物在发酵培养基接种时加入，诱导物浓度为0.2g/L。此时，ADD最高产量为0.5g/L，转化率达到25%，发酵时间为8天。20241
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毕业论文关键词：植物甾醇；分枝杆菌；ADD；雄烯二酮
Preliminary Study on the Process of Manufacture Medication Intermediate ADD Using Plant Steroid
Abstract: 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gradual depletion of diosgenin and other natural resources, the method of producing ADD by chemical synthesis system is no longer applicable. It is well-known that phytosterols are new suitable raw materials for microbial degradation to ADD because of low cost and easy availability. In recent years, studies on preparation of ADD by mycobacteria degradation of plant sterols system have been widely carried out. In this paper, a strain preserved in our laboratory was used as the starting strain and soybean sterols as substrate. The process on production of ADD was explored. The main work of this study was as follows: the choice of carbon source, nitrogensourceof medium, iron ion and its inhibitor, the culture temperature, and adding amount of substrate were investigated. Then the concentrations of carbon source, nitrogen source, time for adding substrate, inducer concentration were optimized by orthogonal test. Preliminary determination of the optimum carbon source of medium was glucose, and its concentration was 7.5 g/L, the optimum nitrogen source was ammonium dihydrogen phosphate, the concentration was 2 g/L, medium cultured with 0.005 g/L ammonium ferric citrate, supplementary iron ions for cell growth, and add 0.025 g/L of six hydrate cobalt chloride before the substrate was added, the most suitable culture temperature was 30 ℃, the optimum substrate adding amount was 2g/L, the optimum substrate addition time was 24h, using AD as the inducer in fermentation culture medium, inducer concentration was 0.2 g/L. At this time, the highest yield of ADD was 0.5g/L, the conversion rate reacheed 25%, fermentation time was 8 days.
KeyWords：phytosterol；Mycobacterium；androstenedione
目录
1 前言 1
1.1 植物甾醇概述 1
1.1.1 植物甾醇理化性质 1
1.1.2 植物甾醇的种类及功能 1
1.1.3 植物甾醇的应用现状 2
1.2 甾体激素药物的发展及现状 2
1.2.1 甾体激素药物概述 3
1.2.2 甾体激素类药物的应用现状 3
1.2.3 雄甾二烯二酮（ADD）的研究及应用 4
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