Determination of enzyme substrate concentration

Details Determination of enzyme substrate concentration Determination of enzyme substrate concentration

1978-12-07

1980-08-19

Naff, David M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving urea or urease

435 14, 435 25, 435183, 435188, 435189, 435195, 435227, G01N 3114, C07G 702

Patent

active

042185351

ABSTRACT:
An enzymatic reagent is provided for use in the kinetic measurement of the concentration of an enzyme substrate present in a sample and possessing the Michaelis-Menten relationship between reaction rate and substrate concentration. The reagent includes an enzyme such as glucose oxidase, uricase or urease for reacting with corresponding enzyme substrates such as glucose, uric acid or urea. In addition, the reagent includes a nonreacting competitive inhibitor of the enzyme. The inhibitor is present in an amount sufficient to improve the linearity of the reaction rate-concentration relationship associated with the enzyme substrate reaction at concentrations which are low relative to the Michaelis-Menten constant, Km, for the particular enzyme substrate reaction. The improved linearity, in turn, improves the accuracy of the kinetic measurements of substrate concentration made by linear output analyzers especially in an upper portion of the predetermined range where the relationship is particularly nonlinear.

REFERENCES:
patent: 3977944 (1976-08-01), Muller-Matthesius et al.
Dixon et al., Enzymes. Academic Press Inc., N.Y., 1958 (pp. 171-181).
Zollner et al., Heparin, Methods of Enzymatic Analysis, Academic Press, N.Y., 1965 (pp. 79-83).
Fritz et al., Protease Inhibitors, Methods of Enzymatic Analysis, Academic Press, Inc. N.Y., 1974 (pp. 1064-1068 and 1085).

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