EVOLUTIONARY IMPLICATIONS OF FEATURES OF AROMATIC AMINO-ACID BIOSYNTHESIS IN THE GENUS ACINETOBACTER

Key enzymes of aromatic amono acid biosynthesis were examined in the genus Acinetobacter. Members of this genus belong to a suprafamilial assemblage of Gram-negative bacteria (denoted Superfamily B) for which a phylogenteic tree based upon oligonucleotide cataloging of 16S rRNa exists. Since the Acinetobacter lineage diverged at an early evolutionary time from other lineages within Superfamily B, examination of aromatic biosynthesis in members of this genus has suppled important clues for the deduction of major evolutionary events leading to the contemporary aromatic pathways that now exist within Superfamily B. Together with Escherichia coli, Pseudomonas aeruginosa and Xanthomonas campestris, four well-spaced lineages have now been studied in comprehensive detail with respect to comparative enzymological features of aromatic amino acid biosynthesis. A. calcoaceticus and A. lwoffi both possess two chorismate mutase isozymes: one a monofunctional isozyme (chorismate mutase-F), and the other (chorismate mutase-P) a component of a bifunctional P-protein (chorismate mutase-prephenate dehydratase). While both P-protein activities were feedback inhibited by L-phenylalanine, the chorismate mutase-P activity was additionally inhibited by prephenate. Likewise, chorismate mutase-F was product inhibited by prephenate. Two isozymes of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase were detected. The major isozyme (> 95%) was sensitive to feedback inhibition to L-tyrosine, whereas the minor isozyme was apparently insensitive to allosteric control. Prephenate dehydrogenase and arogenate dehydrogenase activities were both detected, but could not be chromatographically resolved. Available evidence favors the existence of a single dehydrogenase enzyme, exhibiting substrate ambiguity for prephenate and L-arogenate. Dehydrogenase activity with either of the latter substrates was specific for NADP+, NAD+ being ineffective. Consideration of the phylogney of Superfamily-G organisms suggests that the stem ancestor of the Superfamily possessed a single dehydrogenase enzyme having ambiguity for both substrate and pyridine nucleotide cofactor. Since all other members of Superfamily B have NAD+-specific dehydrogenases, specialization for NADP+-specific dehydrogenases, specialization for NADAP+ must have occurred following the point of Acinetobacter divergence, leading to the dichotomy seen in present-day Superfamily-B organisms.