Abstract

A rate equation for two-substrate enzyme reaction has been proposed by Ingraham & Makower (1954), and by Dixon & Webb (1958). This paper attempts to rearrange the equation for the purpose of practical application. Both the double-reciprocal relation and that of “ a/v versus a” are represented by hyperbolas indicating substrate effects, either substrate-acceleration or substrate-inhibition. The substrate effects appear as a pair of reverse phenomena, that is, substrate-acceleration by one of the two substrates and substrate-inhibition by the other one, according to mathematical expressions of these. A cyclic sequence of the intermediate complexes is indicated by an interrelation of the partial rate-constants. A graphical method to estimate both the maximum rate of the reaction and a dissociation constant of the EAB ternary complex is achieved by plotting 1/ v-ordinates of the double-reciprocal hyperbolas at the 1/ v-axis against the 1/ b-values, reciprocals of the concentrations of the second substrate B.

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