Slow motion picture of protein inactivation during single-droplet drying: a study of inactivation kinetics of L-glutamate dehydrogenase dried in an acoustic levitator.
Slow motion picture of protein inactivation during single-droplet drying: a study of inactivation kinetics of L-glutamate dehydrogenase dried in an acoustic levitator.
复制标题
单滴干燥过程中蛋白质失活的慢动作：声悬浮器中干燥的 L-谷氨酸脱氢酶失活动力学的研究。
DOI：
10.1002/jps.23129
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发表时间：
2012
影响因子：
3.8
通讯作者：
Geoffrey Lee
中科院分区：
文献类型：
--
作者：
E. Lorenzen;Geoffrey Lee
提出了一种新的技术，可以测量在声悬浮单个液滴干燥过程中蛋白质失活的动力学。在干燥过程中，经过不同的时间从声场中去除液滴/颗粒，并分析液滴/颗粒中蛋白质的状态。研究了干燥空气温度、相对湿度、缓冲液浓度和底物存在对谷氨酸脱氢酶失活的影响。失活动力学表现为三个明显的阶段。在恒速干燥的第一阶段，液滴中的蛋白质几乎没有失活。在干燥临界点后，当表面温度急剧上升时，可以区分出第二相，但固体颗粒中的蛋白质仍然只有很少的失活。蛋白质快速失活的起始点标志着第三阶段的开始，该阶段以大约一级速率动力学进行。在L谷氨酸脱氢酶的情况下，证据表明，决定蛋白质失活起点的是固体的残余水分含量，而不是温度本身。
A novel technique is presented to allow measurement of the kinetics of protein inactivation during drying of an acoustically levitated single droplet. Droplets/particles are removed from the acoustic field after various times during drying, and the state of the protein within them is analyzed. The influence of drying air temperature, relative humidity, buffer concentration, and the presence of a substrate on the inactivation of glutamate dehydrogenase is described. The kinetics of inactivation showed three distinct phases. The first phase of constant drying rate demonstrated little protein inactivation in the solution droplet. After the critical point of drying, a second phase was distinguishable when the surface temperature has risen sharply, but there is still only little inactivation of the protein in the solid particle. An onset point of rapid inactivation of the protein marked the start of the third phase that proceeded with approximately first-order rate kinetics. In the case of L-glutamate dehydrogenase, the evidence suggests that the residual moisture content of the solid and not the temperature alone determines the point of onset of protein inactivation.