Ornithine decarboxylase (ODC) from Lactobacillus 30a catalyses the cleavage of α-methylornithine into ammonia and 2-methyl-1-pyrroline; glutamate decarboxylase (GAD) from Escherichia coli catalyses the cleavage of α-methylglutamate into ammonia and laevulinic acid. In our analyses, 2-methyl-1-pyrroline and laevulinic acid were identified by HPLC and mass spectroscopic analysis, and ammonia was identified by means of glutamate dehydrogenase. Molecular oxygen was consumed during these reactions in a 1:2 molar ratio with respect to the products. The catalytic efficiencies (kcat/Km) of the reactions catalysed by ODC and GAD were determined as 12500 and 9163 M-1˙min-1 respectively. When the reactions were performed under anaerobic conditions, no ammonia, 2-methyl-1-pyrroline or laevulinic acid was produced to a significant extent. The formation of ammonia and O2 consumption (in a 1:2 molar ratio with respect to ammonia) were also detected during the reaction of ODC and GAD with putrescine and γ-aminobutyrate respectively. Taken together, these findings clearly indicate that ODC and GAD catalyse an oxidative deamination of their decarboxylation products, a reaction similar to that catalysed by dopa decarboxylase (DDC) with α-methyldopa [Bertoldi, Dominici, Moore, Maras and Borri Voltattorni (1998) Biochemistry 37, 6552-6561]. Furthermore, this reaction was accompanied by a decarboxylation-dependent transamination occurring for GAD, DDC and ODC with a frequency of approx. 0.24%, 1% and 9% respectively compared with that of oxidative deamination.
Ornithine and glutamate decarboxylases catalyse an oxidative deamination of their α-methyl substrates
- Views Icon Views
- Share Icon Share
Mariarita BERTOLDI, Virginia CARBONE, Carla BORRI VOLTATTORNI; Ornithine and glutamate decarboxylases catalyse an oxidative deamination of their α-methyl substrates. Biochem J 15 September 1999; 342 (3): 509–512. doi: https://doi.org/10.1042/bj3420509
Download citation file: