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Lagtime: a program for calculating coupled enzyme assay parameters.

S P Brooks, C H Suelter

    International Journal of Bio-Medical Computing
    |September 1, 1986
    PubMed
    Summary
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    This program calculates enzyme lagtime or required auxiliary enzyme units for coupled enzyme assays. It optimizes assay costs when using two auxiliary enzymes and accounts for intermediate mutarotation.

    Area of Science:

    • Biochemistry
    • Enzyme Kinetics
    • Computational Biology

    Background:

    • Coupled enzyme assays are crucial for determining enzyme activity.
    • Accurate determination of lagtime is essential for reliable kinetic measurements.
    • Auxiliary enzyme concentrations significantly impact assay performance.

    Purpose of the Study:

    • To develop a program for calculating enzyme lagtime or required auxiliary enzyme units.
    • To provide a tool for optimizing coupled enzyme assay conditions.
    • To incorporate mutarotation and cost minimization into enzyme assay calculations.

    Main Methods:

    • Utilized equations from S.P.J. Brooks et al. for coupled enzyme systems.
    • Developed a computational program for kinetic calculations.

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  • Incorporated rate constants for mutarotation of intermediates.
  • Included cost minimization for assays using two auxiliary enzymes.

    • Main Results:

    • The program accurately calculates the time required for observed rates to approximate enzyme rates (lagtime).
    • It determines the necessary units of auxiliary enzymes for a desired lagtime.
    • The program successfully accounts for mutarotation in coupled reactions.
    • Cost optimization for dual auxiliary enzyme assays is achieved.

    Conclusions:

    • The developed program offers a versatile tool for enzyme kinetic studies.
    • It enhances the accuracy and efficiency of coupled enzyme assays.
    • The program's flexibility accommodates complex reaction systems, including those with mutarotation.