<p>In this paper horseradish peroxidase has been employed to study the inhibitor potential of boronic acid derivative 2,4,6-trifluorophenylboronic acid in three concentrations. Temperature and pH effect were also studied to get a closer look on this possible inhibitor behaviour. The activation energy determined for the inhibited and uninhibited reaction gave same results (21.2&#xa0;kJ&#xa0;mol⁻¹ in the absence and 20.7 and 21.2&#xa0;kJ&#xa0;mol⁻¹ in the presence of the inhibitor at concentrations of 8 and 16&#xa0;mM, respectively). Energy activation in the presence of 24&#xa0;mM concentration of inhibitor was significantly lower compared to others, and the Arrhenius plot for this concentration also yielded unusually small value for collision factor. Values of activation energy are similar for uninhibited and also inhibited reactions and it is observed that inhibition percentage was the highest at 37&#xa0;°C. Activation energies values could be explained, that at a given enzyme concentration according to different parameters such as temperature, substrate concentration, and presence of inhibitor usually the apparent velocity of the reaction varies which is mainly dependent on the probability of efficient collision between substrate and enzyme. According to our data Arrhenius constant <i>A</i> is decreased, while the inhibitor concentration is increased, which is in an accordance with the good enzyme–inhibitor relationship “logic”. This type of inhibitor is usually affecting HRP conformation. The activity of enzyme is being reduced consequently. Inhibition of horseradish peroxidase activity by inhibitor 2,4,6-trifluorophenylboronic acid follows a mixed type kinetics. The efficiency of the inhibitor was the concentration dependent.</p> Graphical abstract <p></p>