微生物法合成11α-羟基-ADD的发酵工艺初步研究
关键词：赭曲霉；甾体；11α-羟基化；发酵；ADD
The preliminary study of microbial synthesis of
11α-hydroxy-ADD fermentation process
ABSTRCT: Steroid hormone drug is the second medicine whose production is just inferior to antibiotic. In the production of steroid hormone drug, some groups of steroid need modifying, 11α-hydroxylation is one of the most important ones. Therefore, the study of steroid 11α hydroxylation has the important theory and the practical significance. Microbiology enzymatic reaction is used to translate 1, 4-Androstadiene-3, 17-dione into 11-Hydroxy-1, 4 - Androstadiene-3, 17-dione. In this paper, the content is to optimize the process conditions of Aspergillus Ochraceus’s hydroxylation 1, 4 - Androstadiene-3, 17-dione. Study the influence of Aspergillus Ochraceus’s hydroxylation activity in different medium glucose concentration, substrate concentration, the concentration of starch, the time of Aspergillus Ochraceus’s inoculation and pH, further experiments to determine the optimum conditions orthogonal. Furthermore, orthogonal experiment is used to determine the optimal process conditions. The optimum conditions were: the concentration of glucose 20g/L, substrate concentration 9g/L, starch concentrations of 9g/L, pH pH 7.0.
Keywords: Aspergillus Ochraceus; steroid; 11α-hydroxy; ferment; ADD
目录
1 前言 1
1.1 本课题国内外研究现状概述 1
1.1.1 11α-羟基雄甾-1,4-二烯-3,17-二酮简述 1
1.1.2 雄甾-1,4-二烯-3,17-二酮简述 1
1.1.3 微生物转化合成甾体类药物研究现状 2
1.1.4 甾体微生物转化技术及工艺研究现状 3
1.2 本课题的研究目的和意义 5
2 材料和方法 7
2.1 材料试剂、仪器设备 7
2.1.1 仪器设备 7
2.1.2 材料试剂 7
2.2 检测及其计算方法 7
2.2.1 葡萄糖含量的测定 7
2.2.2 菌体浓度的测定 8
2.2.3 提取产物的方法——萃取 8
2.2.4 ADD、11α-OH ADD的薄层层析检测法（TLC） 8
2.2.5 ADD，11α-OH-ADD高效液相色谱（HPLC）检测法 8
2.2.6 质量转化率、转化效率的计算方法 8
2.3 实验方法 9
2.3.1 试剂制备 9
2.3.2 培养基配方 9
2.2.3 葡萄糖浓度对雄甾-1,4-二烯-3,17-二酮羟基化的影响 9
2.2.4 底物浓度对雄甾-1,4-二烯-3,17-二酮羟基化的影响 10
2.2.5 淀粉浓度对雄甾-1,4-二烯-3,17-二酮羟基化的影响 10
2.2.6 接种时间对雄甾-1,4-二烯-3,17-二酮羟基化的影响 11
2.2.7 pH值对雄甾-1,4-二烯-3,17-二酮羟基化的影响 11
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