Correlation of the kinetics of aggregation and inactivation of L-glutamate dehyrogenase during drying and particle formation of a levitated microdroplet
Correlation of the kinetics of aggregation and inactivation of L-glutamate dehyrogenase during drying and particle formation of a levitated microdroplet
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干燥过程中 L-谷氨酸脱氢酶聚集和失活动力学与悬浮微滴颗粒形成的相关性
DOI：
10.1080/07373937.2018.1445097
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发表时间：
2018
影响因子：
3.3
通讯作者：
Lee. G.
中科院分区：
文献类型：
--
作者：
Lee. G.
声悬浮器中单液滴干燥的测量已用于检测干燥和颗粒形成过程中模型蛋白 L-谷氨酸脱氢酶 (GDH) 的聚集动力学和失活之间的相关性。液滴干燥在一组条件下进行：干燥气体温度≥60℃；干燥气体相对湿度 = 1%，干燥气体流速 = 0.5 L/min。使用离心技术从从悬浮室取出的再水合微粒中去除不溶性聚集物。然后可以使用尺寸排阻色谱来定量非聚集的天然蛋白质的含量。在恒速干燥期间直至干燥临界点，保留了非聚集的天然结构。仅在达到干燥的临界点后，由于不溶性聚集体的形成，才观察到非聚集的天然含量迅速下降。没有发现可溶性 GDH 三聚体的迹象。这种行为的动力学与文献中悬浮微滴中相同酶的失活动力学密切相关，即活性保留到临界点，而在临界点之后活性迅速丧失。这项工作的新颖之处在于，可以在干燥临界点两侧的干燥和颗粒形成过程中检测到聚集和失活之间的这种相关性。
Measurement of single droplet drying in the acoustic levitator has been used to detect a correlation between the kinetics of aggregation and inactivation of the model protein l-glutamate dehydrogenase (GDH) during drying and particle formation. The droplet drying was performed at one set of conditions: drying gas temperature = 60°C; drying gas relative humidity = 1%, and drying gas velocity = 0.5 L/min. A centrifugation technique was used to remove insoluble aggregates from the rehydrated microparticles removed from the levitator chamber. Size exclusion chromatography could then be used to quantify the contents of nonaggregated native protein. During the constant-rate drying period up to the critical point of drying, there was retention of nonaggregated native structure. Only after the critical point of drying has been reached, a rapid decline in nonaggregated native content was observed owing to formation of insoluble aggregates. No sign of a soluble trimer of GDH was found. The kinetics of this behavior correlates closely with the kinetics of inactivation of the same enzyme in levitated microdroplets from the literature, i.e., retention of activity up to the critical point and rapid loss of activity after the critical point. The novelty of this work is that such a correlation between aggregation and inactivation can be detected during drying and particle formation on both sides of the critical point of drying.
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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。
影响因子：
3.8
作者：
E. Lorenzen;Geoffrey Lee
通讯作者：
Geoffrey Lee
DOI：
--
发表时间：
2007
期刊：
影响因子：
--
作者：
H. Yoshii;T-L. Neoh;M. Ohkawara and T.Furuta
通讯作者：
M. Ohkawara and T.Furuta
DOI：
--
发表时间：
1975
期刊：
Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine
影响因子：
--
作者：
K. S. Rogers
通讯作者：
K. S. Rogers
碳水化合物和蛋白质溶液的干燥
DOI：
--
发表时间：
1994
期刊：
影响因子：
--
作者：
Shuichi Yamamoto;Y. Sano
通讯作者：
Y. Sano
脲酶的热诱导失活和聚集：实验和群体平衡模型
DOI：
--
发表时间：
2012
期刊：
影响因子：
--
作者：
Peter Grančič;V. Illeová;M. Polakovič;J. Sefcik
通讯作者：
J. Sefcik