琥珀酸脱氢酶的竞争性抑制作用 综合实验设计

Design on Biochemistry Experiment Teaching of Competitive Inhibition in Succinate Dehydrogenase Activity

  • 摘要: 为加强学生的生物化学实验操作和分析能力,对琥珀酸脱氢酶的竞争性抑制作用进行了综合设计与探索,包括改进酶的提取方法、优化反应体系、确定酶促反应条件、增加实验对照。结果发现:将肌肉研磨且去除红细胞获得的肌肉糜作为改进后的粗酶提取液,无颜色干扰,使酶的竞争性抑制作用现象更直观。固定酶促反应体系中粗酶提取液、琥珀酸和亚甲基蓝的比例,于pH7.4下、37 ℃反应15 min并增设无琥珀酸对照,实验重复性较好。对琥珀酸脱氢酶的竞争性抑制作用实验的综合设计有利于学生理解酶的竞争性抑制作用原理和特点,认识影响酶活性的因素。

     

    Abstract: To strengthen students' experimental operation and experimental design capabilities, the competitive inhibition experiment of succinate dehydrogenase in biochemistry is designed and improved. The redesign includes improving the enzyme extraction method, adding experimental controls, optimizing reaction system, fixing reaction temperature and pH value. To be specific, crude succinate dehydrogenase extract is obtained by removing red blood cells from muscle. The pH value of enzymatic reaction system is 7.4, meanwhile the ratio of succinic acid and methylene blue were fixed and added to enzymatic reaction system. Enzyme reaction is operated at 37 ℃ for 15 minutes with additional succinate free control. In this way the experimental phenomenon of succinate dehydrogenase competitive inhibition becomes more repeatable. The comprehensive design of the experiment is beneficial to understand the principles and characteristics of competitive inhibition of enzymes, and the factors that affect enzyme activity for students.

     

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