通过简单且低成本的实验室活动介绍代谢控制分析,Journal of Chemical Education

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通过简单且低成本的实验室活动介绍代谢控制分析
Journal of Chemical Education ( IF 2.5 ) Pub Date : 2023-11-08 , DOI: 10.1021/acs.jchemed.3c00384
Nicolás Andrés Saffioti _{1,

2,

3} , Carla Angelani ₁ , José María Delfino _{1,

3} , Silvia Fernández Villamil _{1,

4} , Gabriela Elena Gómez _{1,

3} , Irene Mangialavori _{1,

3} , María Florencia Pignataro _{1,

5} , Pablo Julio Schwarzbaum _{1,

3}

Affiliation

代谢控制分析 (MCA) 是一种系统生物学方法，用于测量代谢步骤对代谢途径总体通量的控制。在生物化学本科课程中，可以通过展示 MCA 理论实际应用的实验活动来鼓励 MCA 的学习。在这项工作中，我们提出了一项实验室活动，旨在通过量化密封注射器中CO _{2 的产生来测量}酿酒酵母的糖酵解通量。在不同浓度的聚维酮碘（一种糖酵解抑制剂）下测量糖酵解通量。然后，学生分析实验结果，计算糖酵解通量对抑制剂聚维酮碘的响应系数和敏感性。2022年88名本科生顺利完成了此次活动，提交了实验室报告进行修改。对学生报告的评估表明，该活动鼓励了 MCA 所需技术程序的学习，并有助于识别学生经常持有的误解。实验方法的简单性以及无害且低成本试剂的可用性使学生能够专注于 MCA 框架内理论概念的应用。

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Introduction to Metabolic Control Analysis by a Simple and Low-Cost Laboratory Activity

Metabolic Control Analysis (MCA) is a systems biology approach that measures the control of a metabolic step on the overall flux of a metabolic pathway. In undergraduate courses in biochemistry, learning of MCA can be encouraged by experimental activities demonstrating practical applications of the MCA theory. In this work, we present a laboratory activity designed to measure the glycolytic flux of Saccharomyces cerevisiae by quantifying the CO₂ generation in hermetically sealed syringes. Glycolytic flux was measured at different concentrations of povidone-iodine, an inhibitor of glycolysis. Then, the students analyzed the experimental results to calculate the response coefficient and sensitivity of the glycolytic flux to the inhibitor povidone-iodine. The activity was satisfactorily carried out by 88 undergraduate students in 2022 who submitted a laboratory report for revision. Evaluation of students’ reports indicated that the activity encouraged the learning of technical procedures required for MCA and facilitated the identification of misconceptions frequently held by students. The simplicity of the experimental approach and the availability of harmless and low-cost reagents allow students to focus on the application of theoretical concepts within the MCA framework.

更新日期：2023-11-08

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