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Enzyme-Catalyzed Hydrolysis of Lipids in Immiscible Microdroplets Studied by Contained-Electrospray Ionization
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-09-15 , DOI: 10.1021/acs.analchem.1c02785 Benjamin J Burris 1 , Abraham K Badu-Tawiah 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-09-15 , DOI: 10.1021/acs.analchem.1c02785 Benjamin J Burris 1 , Abraham K Badu-Tawiah 1
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Enzyme-catalyzed hydrolysis of lipids was monitored directly in immiscible microdroplet environments using contained-electrospray mass spectrometry. Aqueous solution of the lipase enzyme from Pseudomonas cepacia and the chloroform solution of the lipids were sprayed from separate capillaries, and the resultant droplets were merged within a reaction cavity that is included at the outlet of the contained-electrospray ionization source. By varying the length of the reaction cavity, the interaction time between the enzyme and its substrate was altered, enabling the quantification of reaction product as a function of time. Consequently, enhancement factors were estimated by comparing rate constants derived from the droplet experiment to rate constants calculated from solution-phase conditions. These experiments showed enhancement factors greater than 100 in favor of the droplet experiment. By using various lipid types, two possible mechanisms were identified to account for lipase reactivity in aerosols: in-droplet reactions for relatively highly soluble lipids and a droplet coalescence mechanism that allows interfacial reactions for the two immiscible systems.
中文翻译：
含电喷雾电离研究不混溶微滴中脂质的酶催化水解
使用包含的电喷雾质谱法在不混溶的微滴环境中直接监测酶催化的脂质水解。洋葱假单胞菌脂肪酶的水溶液脂质的氯仿溶液从单独的毛细管中喷出，所得液滴在包含在封闭式电喷雾电离源出口处的反应腔内合并。通过改变反应腔的长度，酶与其底物之间的相互作用时间发生了变化，从而能够将反应产物定量为时间的函数。因此，通过比较从液滴实验得出的速率常数与从溶液相条件计算的速率常数来估计增强因子。这些实验显示出大于 100 的增强因子有利于液滴实验。通过使用各种脂质类型，确定了两种可能的机制来解释气溶胶中的脂肪酶反应性：
更新日期：2021-09-28
中文翻译：
含电喷雾电离研究不混溶微滴中脂质的酶催化水解
使用包含的电喷雾质谱法在不混溶的微滴环境中直接监测酶催化的脂质水解。洋葱假单胞菌脂肪酶的水溶液脂质的氯仿溶液从单独的毛细管中喷出，所得液滴在包含在封闭式电喷雾电离源出口处的反应腔内合并。通过改变反应腔的长度，酶与其底物之间的相互作用时间发生了变化，从而能够将反应产物定量为时间的函数。因此，通过比较从液滴实验得出的速率常数与从溶液相条件计算的速率常数来估计增强因子。这些实验显示出大于 100 的增强因子有利于液滴实验。通过使用各种脂质类型，确定了两种可能的机制来解释气溶胶中的脂肪酶反应性：
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