An Improved Method for Studying the Enzyme-Catalyzed Oxidation of Glucose Using Luminescent Probes

A new method is presented for measuring the rate of the oxidation of glucose in the presence of glucose oxidase. The improved method employs luminescence measurements to directly determine the concentration of oxygen in real time, thus obviating complicated reaction schemes employed in previous meth...

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Bibliographic Details
Published in:Journal of chemical education 2007-09, Vol.84 (9), p.1511
Main Authors: Cuber, Matthew, Demas, J. N, Bare, William D, Pham, Chi V
Format: Article
Language:English
Subjects:
Biochemistry
Catalytic oxidation
Chemical reactions
Chemistry
Glucose
Kinetics
Laboratories
Laboratory Equipment
Laboratory Procedures
Laboratory tests
Lighting
Measurement Equipment
Measurement Techniques
Methods
Oxidation
Probes
Reaction kinetics
Science Activities
Science education
Science Instruction
Science Laboratories
Teaching Methods
Undergraduate Study
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Summary:A new method is presented for measuring the rate of the oxidation of glucose in the presence of glucose oxidase. The improved method employs luminescence measurements to directly determine the concentration of oxygen in real time, thus obviating complicated reaction schemes employed in previous methods. Our method has been used to determine kinetic parameters including V max, K m, and k cat, all in good agreement with published values. The method can also be used to investigate specificity and inhibition. Information is included pertaining to the construction of a low-cost fluorescence instrument suitable for the measurements in this protocol.
ISSN:0021-9584
1938-1328
DOI:10.1021/ed084p1511