Single-Shot Characterization of Enzymatic Reaction Constants K m and k cat by an Acoustic-Driven, Bubble-Based Fast Micromixer

Yuliang Xie; Daniel Ahmed; Michael Ian Lapsley; Sz-Chin Steven Lin; Ahmad Ahsan Nawaz; Lin Wang; Tony Jun Huang

doi:10.1021/ac301590y

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Single-Shot Characterization of Enzymatic Reaction Constants K m and k cat by an Acoustic-Driven, Bubble-Based Fast Micromixer

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Single-Shot Characterization of Enzymatic Reaction Constants K m and k cat by an Acoustic-Driven, Bubble-Based Fast Micromixer

Yuliang Xie, Daniel Ahmed, Michael Ian Lapsley, Sz-Chin Steven Lin, Ahmad Ahsan Nawaz, Lin Wang and Tony Jun Huang

Analytical chemistry (Washington), Vol.84(17), pp.7495-7501

09/04/2012

DOI: 10.1021/ac301590y

PMID: 22880882

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Abstract

In this work we present an acoustofluidic approach for rapid, single-shot characterization of enzymatic reaction constants K m and k cat. The acoustofluidic design involves a bubble anchored in a horseshoe structure which can be stimulated by a piezoelectric transducer to generate vortices in the fluid. The enzyme and substrate can thus be mixed rapidly, within 100 ms, by the vortices to yield the product. Enzymatic reaction constants K m and k cat can then be obtained from the reaction rate curves for different concentrations of substrate while holding the enzyme concentration constant. We studied the enzymatic reaction for β-galactosidase and its substrate (resorufin-β-D-galactopyranoside) and found K m and k cat to be 333 ± 130 μM and 64 ± 8 s–1, respectively, which are in agreement with published data. Our approach is valuable for studying the kinetics of high-speed enzymatic reactions and other chemical reactions.

Details

Title: Subtitle: Single-Shot Characterization of Enzymatic Reaction Constants K m and k cat by an Acoustic-Driven, Bubble-Based Fast Micromixer
Creators: Yuliang Xie - Pennsylvania State University
Daniel Ahmed - Pennsylvania State University
Michael Ian Lapsley - Pennsylvania State University
Sz-Chin Steven Lin
Ahmad Ahsan Nawaz - Pennsylvania State University
Lin Wang - Ascent Bio-Nano Technologies
Tony Jun Huang - Pennsylvania State University
Resource Type: Journal article
Publication Details: Analytical chemistry (Washington), Vol.84(17), pp.7495-7501
Publisher: American Chemical Society
DOI: 10.1021/ac301590y
PMID: 22880882
ISSN: 0003-2700
eISSN: 1520-6882
Language: English
Date published: 09/04/2012
Academic Unit: Roy J. Carver Department of Biomedical Engineering
Record Identifier: 9984196987202771

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