An electrometric method for the determination of tyrosinase activity.

The pathway of dopachrome formation from L-dopa involves the net release of one proton for each molecule of dopachrome formed. The protons produced as a consequence of the enzymic step catalysed by tyrosinase can be measured by an electrometric device able to monitor changes in H+ concentration below 1 microM. This electrometric recording can be used as a simple, sensitive and continuous method for determining tyrosinase activity.

Inhibition of enzyme-catalysed reactions by excess substrate. A theoretical and Monte Carlo study of turning points in v(S) graphs.

Some difficulties associated with observing and defining substrate inhibition curves are discussed. Then a new method for measuring the steepness of substrate inhibition curves is derived and a parameter, lambda, is defined to measure the steepness of descent from a maximum or steepness of ascent from a minimum in v(S) plots. A mathematical theorem is presented to show how the magnitude of lambda is limited by the degree of the rate equation.

Non-proteolytic solubilization of bovine thyroid peroxidase: thermodynamic parameters of the thermoinactivation.

1. A subcellular fractionation from bovine thyroid gland homogenate was carried out by differential centrifugation. The maximal peroxidase activity was found in the microsomal fraction.

Steady-state study of the mechanism of dopa-oxidase activity of tyrosinase.

The mechanism of the dopa-oxidase activity of frog epidermis tyrosinase has been studied. Initial reaction rates have been measured as function of substrate concentrations, L-dopa and oxygen, in the presence and absence of an inhibitor, product of the reaction. Initial reaction rates versus substrate concentrations, without inhibitor, show a linear dependence in the double-reciprocal space, that discarded Ordered and Random mechanisms.