化学生物学实验“酶米氏常数测定”的改进与创新  

Improvements and innovations of chemical biology experiment“michaelis constant determination of enzyme”

作　　者：包婷[1] 高雅 伍珍[1] 段瑞雪 熊成义 翁玥 何汉平[1] 王升富[1] Bao Ting;Gao Ya;Wu Zhen;Duan Rui-xue;Xiong Cheng-yi;Weng Yue;He Han-ping;Wang Sheng-fu(College of Chemistry and Chemical Engineering,Hubei University,Wuhan 430062,China)

机构地区：[1]湖北大学化学化工学院,湖北武汉430062

出　　处：《化学传感器》2024年第2期62-66,共5页Chemical Sensors

基　　金：湖北大学2022年度教学改革研究项目“基于SPOC线上线下混合式教学的地方性高校化学类综合实验的重塑和优化”(编号2022008);湖北省高等学校省级教学研究项目“以问题为导向的分析化学开放式实验教学模式研究与实践”(编号206)的支持

摘　　要：酶米氏常数测定是化学生物学一个经典的实验项目,通过测定酶催化过程的动力学曲线来评估酶对底物的亲和力及催化活性。通常在实验教学中使用天然酶进行催化反应,如辣根过氧化物酶、过氧化氢酶、碱性磷酸酶等,尽管具有反应高效、底物专一、反应条件温和等优点,但天然酶的成本较高,且其活性易受外界环境影响而导致催化能力降低。为了解决上述问题,该文提出利用二硫化铁(FeS_(2))纳米酶代替传统的天然酶进行催化反应并测定其米氏常数。与天然酶相比,纳米酶不仅具备纳米材料的独特性能,又能够模拟天然酶的催化活性,具有易于合成、催化效率高、稳定性好、成本低及可规模化制备等突出优势。通过指导学生合成不同结构、形貌和催化活性的纳米酶并测定其米氏常数,有助于学生深入学习纳米酶的合成、表征和测试方法,促进学生对纳米材料研究前沿的理解和掌握。Michaelis constant determination experiment of enzyme is a classical experimental project in chemical biology,which is used to assess the affinity and catalytic activity towards a substrate by determining the kinetic curve of the enzyme catalytic process.Usually,natural enzymes including horseradish peroxidase,catalase,alkaline phosphatase are used for the catalytic reactions in experimental teaching.Although natural enzymes hold the advantages of high reaction efficiency,substrate specificity and mild reaction conditions,however,the cost of natural enzymes is high and they are susceptible to the influence of the external environment,leading to the decrease of catalytic ability.In order to solve the above problem,this paper proposes to use nanozymes instead of traditional natural enzymes to catalyze the reaction and determine the Michaelis constant.Compared with natural enzymes,nanozymes not only possess the unique properties of nanomaterials,but also can mimic the catalytic activity of natural enzymes,which hold the outstanding advantages of easy synthesis,high catalytic efficiency,good stability,low cost and scalable preparation.By instructing students to synthesize nanozymes with different structures,morphologies and catalytic activities and determine the Michaelis constant of nanozymes,which can contribute to the in-depth learning about the synthesis,characterization and testing methods of nanozymes,and promotes their understanding and mastery for the research frontiers of nanomaterials.

关 键 词：FeS_(2)纳米酶 结构调控 催化活性 米氏常数 实验改进与创新 

分 类 号：G642[文化科学—高等教育学] TQ426.97-4[文化科学—教育学]