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Determination of Kinetic Parameters, Km and kcat, with a Single Experiment on a Chip

Title
Determination of Kinetic Parameters, Km and kcat, with a Single Experiment on a Chip
Author
Jong Wook Hong
Keywords
Reagents; Chemical reactions; Biotechnology; Peptides and proteins; Fluid dynamics
Issue Date
2009-04
Publisher
AMER CHEMICAL SOC
Citation
ANALYTICAL CHEMISTRY, v. 81, No. 9, Page. 3239-3245
Abstract
We have demonstrated a multistep enzyme reaction on a chip to determine the key kinetic parameters of enzyme reaction. We designed and fabricated a fully integrated microfluidic chip to have sample metering, mixing, and incubation functionalities. The chip generates a gradient of reagent concentrations in 11 parallel processors. We used β-galactosidase and its substrate, resorufin-β-d-galactopyranoside, as the model system of the enzyme reaction. With a single experiment on the chip, we determined the key parameters for the enzyme kinetics, Km and kcat, and evaluated the effect of inhibitor concentrations on the reaction rates. This study provides a new tool for evaluating various effectors, such as inhibitors and cofactors, on the initial rate of an enzyme reaction, and it could be applied to a comprehensive bio/chemical reaction study.
URI
https://pubs.acs.org/doi/abs/10.1021/ac8020938https://repository.hanyang.ac.kr/handle/20.500.11754/166485
DOI
10.1021/ac8020938.s002
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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