Label-free direct visual analysis of hydrolytic enzyme activity using aqueous two-phase system droplet phase transitions.
Label-free direct visual analysis of hydrolytic enzyme activity using aqueous two-phase system droplet phase transitions.
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DOI：
10.1021/ac500657k
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发表时间：
2014-04-15
影响因子：
7.4
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Lai, David;Frampton, John P.;Tsuei, Michael;Kao, Albert;Takayama, Shuichi
利用葡聚糖水解法将聚乙二醇(PEG)-葡聚糖(DEX)双水相体系液滴转化为单相，可直接显示葡聚糖酶的活性。DEX液滴是通过手动微移液或在连续的聚乙二醇相中通过计算机控制的连接由背面扩散光光刻形成的圆形通道的孔的驱动而形成的。两相到单相转变所需的时间取决于葡聚糖酶的浓度、介质的pH和温度。根据已报道的催化常数、各温度下聚乙二醇-地塞米松相变的二节点聚合物浓度曲线和pH条件，估算了葡聚糖酶的表观米氏常数。微流控液滴系统与相变观测相结合，为酶活性的无标记直接测量提供了一种新的方法。
Dextran hydrolysis-mediated conversion of polyethylene glycol (PEG)-dextran (DEX) aqueous two-phase system droplets to a single phase was used to directly visualize Dextranase activity. DEX droplets were formed either by manual micropipetting or within a continuous PEG phase by computer controlled actuation of an orifice connecting rounded channels formed by backside diffused light lithography. The time required for the two-phase to one-phase transition was dependent on the Dextranase concentration, pH of the medium, and temperature. The apparent Michaelis constants for Dextranase were estimated based on previously reported catalytic constants, the binodal polymer concentration curves for PEG-DEX phase transition for each temperature, and pH condition. The combination of a microfluidic droplet system and phase transition observation provides a new method for label-free direct measurement of enzyme activity.
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影响因子：
--
作者：
Albertsson, P A
通讯作者：
Albertsson, P A
影响因子：
6.1
作者：
Basu, Amar S.
通讯作者：
Basu, Amar S.
影响因子：
6.1
作者：
Lai, David;Frampton, John P.;Takayama, Shuichi
通讯作者：
Takayama, Shuichi
影响因子：
3.3
作者：
El-Tanash, A. B.;El-Baz, E.;Sherief, A. A.
通讯作者：
Sherief, A. A.
DOI：
10.1073/pnas.0404353101
发表时间：
2004-11-09
影响因子：
11.1
作者：
Gu, W;Zhu, XY;Takayama, S
通讯作者：
Takayama, S