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Using the Glucose Oxidase/Peroxidase System in Enzyme Kinetics

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Robert C. Bateman Jr. and Jeffrey A. Evans

Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406

J. Chem. Educ., 1995, 72 (12), p A240

DOI: 10.1021/ed072pA240

Publication Date: December 1995

Abstract

An expeimental protocol is described for a biochemistry laboratory on enzyme kinetics using glucose oxidase coupled to a horseradish peroxidase/colored dye system to yield an easily visualized and quantitated product. The experiment is designed to illustrate the concepts of specific activity, substrate specificity and kinetic parameters. Optional extensions of the protocol could explore parameters of coupled enzyme systems, reversible inhibition and enzyme immobilization.

Keywords (Audience):

Upper-Division Undergraduate

Keywords (Domain):

Biochemistry

Keywords (Feature):

The Modern Student Laboratory

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Enzymes

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Citing Articles

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This article has been cited by 11 ACS Journal articles (5 most recent appear below).

A Comprehensive Enzyme Kinetic Exercise for Biochemistry
Janice S. Barton
Journal of Chemical Education2011 Article ASAP
- A Comprehensive Enzyme Kinetic Exercise for Biochemistry
  Janice S. Barton
  Journal of Chemical Education2011 Article ASAP
  This article describes a comprehensive treatment of experimental enzyme kinetics strongly coupled to electronic data acquisition and use of spreadsheets to organize data and perform linear and nonlinear least-squares analyses, all in a manner that ...
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links
Homemade Bienzymatic-Amperometric Biosensor for Beverages Analysis
M. C. Blanco-López , M. J. Lobo-Castañón and A. J. Miranda-Ordieres
Journal of Chemical Education2007 84 (4), 677
- Homemade Bienzymatic-Amperometric Biosensor for Beverages Analysis
  M. C. Blanco-López , M. J. Lobo-Castañón and A. J. Miranda-Ordieres
  Journal of Chemical Education2007 84 (4), 677
  In this experiment students construct an amperometric glucose biosensor and use it to determine the glucose concentration in soft drinks. This experiment is appropriate for an advanced instrumental course; however, it could be adapted to elementary ...
  Abstract | Hi-Res PDF | PDF w/ Links
Advantages of the Pre-Immobilization of Enzymes on Porous Supports for Their Entrapment in Sol−Gels
Lorena Betancor, Fernando López-Gallego, Aurelio Hidalgo, Manuel Fuentes, Ondrej Podrasky, Gabriela Kuncova, José M. Guisán, and Roberto Fernández-Lafuente
Biomacromolecules2005 6 (2), 1027-1030
- Advantages of the Pre-Immobilization of Enzymes on Porous Supports for Their Entrapment in Sol−Gels
  Lorena Betancor, Fernando López-Gallego, Aurelio Hidalgo, Manuel Fuentes, Ondrej Podrasky, Gabriela Kuncova, José M. Guisán, and Roberto Fernández-Lafuente
  Biomacromolecules2005 6 (2), 1027-1030
  In this work, we have compared the entrapment of free or previously immobilized glucose oxidase using a solgel technique. The preimmobilization was carried out on Sepabeads (a porous support) derivatized with glutaraldehyde as the functional group. The ...
  Abstract | Full Text HTML | Hi-Res PDF
Application of Datalogger in Biosensing: A Glucose Biosensor
Martin M. F. Choi and Pui Shan Wong
Journal of Chemical Education2002 79 (8), 982
- Application of Datalogger in Biosensing: A Glucose Biosensor
  Martin M. F. Choi and Pui Shan Wong
  Journal of Chemical Education2002 79 (8), 982
  This article describes the use of a datalogger in conjunction with an oxygen sensor to fabricate a glucose biosensor. Glucose oxidase is first covalently immobilized on an eggshell membrane. The membrane is then used to cover the surface of an oxygen ...
  Abstract | Hi-Res PDF | PDF w/ Links
Factors affecting reaction kinetics of glucose oxidase
Kristin A. Johnson , Beth A. Kroa and Tony Yourey
Journal of Chemical Education2002 79 (1), 74
- Factors affecting reaction kinetics of glucose oxidase
  Kristin A. Johnson , Beth A. Kroa and Tony Yourey
  Journal of Chemical Education2002 79 (1), 74
  Basic principles of enzyme kinetics are demonstrated using the enzyme glucose oxidase. The glucose oxidase enzymatic reaction is coupled to horseradish peroxidase, which in turn catalyzes the oxidation of a dye to a bright blue-green color. The appearance ...
  Abstract | Hi-Res PDF | PDF w/ Links