期刊
ELECTROPHORESIS
卷 41, 期 3-4, 页码 175-182出版社
WILEY
DOI: 10.1002/elps.201900334
关键词
Capillary electrophoresis; Grapheme oxide; Immobilized enzyme microreactor; L- lactate dehydrogenase; Pyruvate
资金
- National Natural Science Foundation of China [21775017]
- Natural Science Foundation of Jilin Province, China [20180101174JC]
- Jilin Provincial Department of Education and Jilin Provincial Key Laboratory of Micro-Nano Functional Materials (Northeast Normal University)
We report the first application of hydrophobic interaction between graphene oxide (GO) and negatively charged enzymes to fabricate CE-integrated immobilized enzyme microreactors (IMERs) by a simple and reliable immobilization procedure based on layer by layer assembly. L-lactate dehydrogenase (L-LDH), which is negatively charged during the enzymatic reaction, is selected as the model enzyme. Various spectroscopic techniques, including SEM, FTIR, and UV-vis are used to characterize the fabricated CE-IMERs, demonstrating the successful immobilization of enzymes on the negatively charged GO layer in the capillary surface. The IMER exhibits excellent repeatability with RSDs of inter-day and batch-to-batch less than 3.49 and 6.37%, respectively, and the activity of immobilized enzymes remains about 90% after five-day usage. The measured K-m values of pyruvate and NADH of the immobilized L-LDH are in good agreement with those obtained by free enzymes. The results demonstrate that the hydrophobic interactions and/or pi-pi stacking is significant between the GO backbone and the aromatic residues of L-LDH and favorable to fabrication of CE-integrated IMERs. Finally, the method is successfully applied to the determination of pyruvate in beer samples.
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