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2.4. Biotransformation condition and product isolation For each fungal species, ten 250 mL Erlenmeyer flasks, each containing 150 mL of sterilized liquid medium were inoculated with freshly obtained spores from agar slope cultures. The flasks were incubated at 26 ?C under constant shaking on an orbital shaker at 120 rpm. After 3 days of growth, 100 mg of substrate dissolved in 1 mL of absolute ethanol was added to each of the 150 mL cultures. Incubations were continued and the progress of the reaction was monitored by TLC. The cultures were generally sampled every day after addition of substrate. In order to study the catalytic efficiencyof microorganisms, substrate controls consisted of sterile medium were similarly incubated but without microorganism. At the end of the incubation period, the products were filtered and extracted three times with 20 mL of chloroform. After evaporating the organic solvent layer, each of the extracts was further purified separately. |
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jiangguofeng
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bxzc123(RXMCDM代发): 金币+20, 多谢应助！ 2015-07-14 09:03:17
bxzc123(RXMCDM代发): 金币+20, 多谢应助！ 2015-07-14 09:03:17
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2.4. 生物转化条件和产物分离 对于每种真菌,每个装有150mL经灭菌的液体培养基的250毫升锥形烧瓶十个,将刚刚由琼脂斜面培养物得到的孢子分别注入每个烧瓶中。烧瓶于一个轨道摇床上以120rpm不断振摇下26℃中培育。培养三天之后,将溶于1mL无水乙醇中的100mg底物加至每个150 mL培养物中。继续培育，通过薄层色谱法监控反应进程。添加底物后每天对培养物进行常规取样。为了研究微生物的催化效率,由无菌培养基组成的底物对照物也同样地进行培育，只是不加微生物。在培育期结束时，过滤产物并用20 mL氯仿提取三次。 有机溶剂层经蒸发后,进一步分别精制每个提取物。 |
2楼2015-03-30 12:04:23
