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bioRxiv · 10.1101/2022.12.15.520621

Derivation of Steady-State First-order Rate Constant Equations for Enzyme-Substrate Complex Dissociation, as well as Zero-order Rate Constant Equations in Relation to Background Assumptions

Udema, I. I.
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Abstract

The maximum velocity (Vmax) of catalysis and the substrate concentration ([ST]) at half the Vmax, the KM, are regarded as steady-state (SS) parameters even though they are the outcomes of zero-order kinetics (ZOK). The research was aimed at disputing such a claim with the following objectives: To: 1) carry out an overview of issues pertaining to the validity of assumptions; 2) derive the needed steady-state (SS) equations distinct from Michaelian equations that can be fitted to both experimental variables and kinetic parameters; 3) calculate the SS first-order rate constant for the dissociation of enzyme-substrate complex (ES) to free substrate, S and enzyme, E; 4) derive the equation of rate constant as a function of the reciprocal of the duration of each catalytic event in the reaction pathway. The experimental values of the data were generated by Bernfeld and Lineweaver-Burk methods. The calculated SS 1st order-order rate constant was << the zero-order Michaelian value, and the difference is {approx} 97.59 % of the zero-order value; the SS catalytic rate differed from the zero-order catalytic rate by {approx} 76.41 % of the latter value; and it was {approx} 93.87 % with respect to the 2nd order rate constant for the formation of enzyme-substrate complex. The equations of time-dependent rate constants, KM, and dissociation constants were derived. The concentration [ST] of the S must be > the concentration ([E0]) of the E for the quasi-steady-state assumption (or approximation) to hold. The SS kinetic parameters are not equivalent to zero-order parameters.

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Udema, I. I.. 2022-12-16. Derivation of Steady-State First-order Rate Constant Equations for Enzyme-Substrate Complex Dissociation, as well as Zero-order Rate Constant Equations in Relation to Background Assumptions. https://doi.org/10.1101/2022.12.15.520621

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