a Institute of Physical Chemistry , PAS , Warsaw , Poland.
b Institute of Organic Chemistry , PAS , Warsaw , Poland.
Prep Biochem Biotechnol. 2019;49(7):727-734. doi: 10.1080/10826068.2019.1605527. Epub 2019 Apr 24.
The accurate estimation of kinetic parameters is of fundamental importance for biochemical studies for research and industry. In this paper, we demonstrate the application of a modular microfluidic system for execution of enzyme assays that allow determining the kinetic parameters of the enzymatic reactions such as - the maximum rate of reaction and - the Michaelis constant. For experiments, the fluorogenic carbonate as a probe for a rapid determination of the kinetic parameters of hydrolases, such as lipases and esterases, was used. The microfluidic system together with the method described yields the kinetic constants calculated from the concentration of enzymatic product changes via a Michaelis-Menten model using the Lambert function . This modular microfluidic system was validated on three selected enzymes (hydrolases).
准确估计动力学参数对于研究和工业的生化研究至关重要。在本文中,我们展示了一种模块化微流控系统在执行酶测定中的应用,该系统允许确定酶反应的动力学参数,例如最大反应速率和米氏常数。对于实验,使用了荧光碳酸盐作为探针,用于快速确定脂肪酶和酯酶等水解酶的动力学参数。微流控系统与所描述的方法一起使用,通过使用 Lambert 函数从酶产物浓度变化计算出米氏-门登模型计算得到的动力学常数。该模块化微流控系统已在三种选定的酶(水解酶)上进行了验证。