青霉素V钾的有机半合成及其抑菌效果表征——推荐一个化学生物学综合实验

Improving the Semi-Synthesis and Antibacterial Study of Penicillin V Potassium as a Chemical Biology Comprehensive Experiment

青霉素的有机半合成及其抑菌效果表征是一个涵盖知识点众多的综合性实验,目前相关实验仅有美国麻省理工学院(MIT)等少数高校开设,国内尚未有关于该实验教学方面的报道。为了将研究型的环境引入教学实验,培养学生科学思维能力,我们进行了如下改进:1)设计并搭建了循环冷凝装置,引入低温控制,提高产物的稳定性,有效减少副产物的生成;2)添加无水乙醇后处理步骤,利用溶解度差异除去副产物;3)引入微量二倍稀释法测定青霉素最低抑菌浓度,利用牛津杯法测定青霉素抑菌圈,提高实验的展示度;4)将科研成果融入教学,使用小麦胚芽凝集素荧光标记结合荧光共聚焦显微镜表征青霉素对细菌细胞壁完整度的影响,展示青霉素的抑菌机制。该实验交叉融合有机合成与化学生物学的实验内容,引入科研表征手段,使得学生切实体验科学研究的过程,激发学生的探索精神和创新意识。

The semi-synthesis and antibacterial study of penicillin V potassium is primarily an interdisciplinary experiment.Currently,only a few foreign universities such as the Massachusetts Institute of Technology(MIT)allow undergraduate students to conduct this experiment.To incorporate a research-like environment into teaching experiments and enable students to develop scientific thinking,the following improvements were made to the experiment:1)A circulating condensing device was designed and built,and low-temperature control was introduced to improve the stability of the products and effectively reduce the formation of by-products.2)Absolute ethyl alcohol was added to remove the by-products based on solubility difference.3)The minimum inhibitory concentration of penicillin was determined using the microbroth dilution method,and antibiotic sensitivity was determined using the Oxford penicillin cup method.4)The effects of penicillin on the integrity of bacterial cell walls were characterized via wheat germ agglutinin fluorescent labeling combined with confocal fluorescence microscopy.This comprehensive experiment,which combines several important multidisciplinary concepts,can develop students’skills in integrated analysis and increase their interest in scientific research.