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Demonstration of the principles of enzyme‐catalysed reactions using alkaline phosphatase
Author(s) -
C.Price Nicholas,
Newman Bethan L.
Publication year - 2000
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1111/j.1539-3429.2000.tb00147.x
Subject(s) - alkaline phosphatase , substrate (aquarium) , enzyme , chemistry , reaction rate , enzyme assay , phosphatase , biochemistry , chromatography , catalysis , biology , ecology
The activity of the enzyme alkaline phosphatase can be conveniently measured spectrophotometrically using 4‐nitrophenylphos‐phate as substrate. This article describes an integrated series of experiments involving the enzyme in which a number of valuable experimental and analytical skills can be developed. A suitable assay procedure for the enzyme is established and this is then used to explore a number of features of the enzyme‐catalysed reaction. These include the dependence of the rate of reaction on substrate concentration, pH and presence of inhibitors. Studies of the temperature‐dependence of the reaction rate in the presence and absence of alkaline phosphatase provide valuable insights into the degree of rate enhancement brought about by the enzyme. © 2000 IUBMB. Published by Elsevier Science Ltd. All rights reserved.