In summer, the concentrations of dissolved organic nitrogen compounds
are often higher than that of inorganic nitrogen. Under such
conditions it would be advantageous if phytoplankton species could utilize
organic nitrogen sources, including free or combined amino
acids, in addition to inorganic nitrogen. This study focused on histidine,
the degradation of which potentially yields three nitrogen
atoms for each molecule of histidine. In this work, histidase from Dunaliella
tertiolecta, a deaminating enzyme catalysing the first steps of
histidine degradation, was purified 4000-fold and partially characterized.
The molecular weight of the native enzyme was estimated to be
155 kDa, corresponding to four subunits of 38 kDa. D. tertiolecta
histidase is stable in the presence of dithiothreitol and is inactivated
by
cyanide. Histidinol phosphate, histidine and Mn2+ are effective
protectors against cyanide inactivation. The enzyme did not exhibit
classical Michaelis-Menten kinetics but showed a relationship between the
rate of catalysis (V) and the concentration of substrate (S)
that
was characteristic of negative allosteric behaviour. A Hill coefficient
of 4 was measured for histidine concentrations higher than 22.5 mM.
Guanine, xanthine and cytosine nucleotides are inhibitors of D. tertiolecta
histidase.