title: Catabolism of aromatic biogenic amines by Pseudomonas aeruginosa PAO1 via meta cleavage of homoprotocatechuic acid.
abstract: Pseudomonas aeruginosa PAO1 catabolized the aromatic amines tyramine and octopamine through 4-hydroxyphenylacetic acid and 3,4-dihydroxyphenylacetic acid (HPA). meta ring cleavage was mediated by 3,4-dihydroxyphenylacetate 2,3-dioxygenase (HPADO), producing 2-hydroxy-5-carboxymethylmuconic semialdehyde (MSA). An NAD-dependent dehydrogenase caused the disappearance of the yellow MSA product, probably forming 2-hydroxy-5-carboxymethylmuconic acid. Induction studies with extracts from mutant cells indicated that the inducer of HPADO was HPA and/or MSA. Strains PAO1.221 (tynC1) and PAO1.303 (tynD1) have chromosomal mutations causing a deficiency in the activity necessary for conversion of 4-hydroxyphenylacetic acid to HPA. Genetic analyses showed that the mutations were in different loci. Strains PAO1.197 (tynE1) and PAO1.185 (tynF1) are deficient in HPADO and the NAD-dependent dehydrogenase, respectively. Plasmid pRO1853 was constructed by cloning approximately 7.3 kilobases of PAO1 chromosomal DNA into the BamHI site of the vector plasmid pRO1614. This recombinant plasmid complemented the tynD1, tynE1, and tynF1 mutations. A putative repressor-binding site involved in the regulation of HPADO synthesis was observed for a subcloned fragment of pRO1853. This recombinant plasmid, pRO1863, failed to complement tynE1 or tynF1 but still complemented tynD1. Another construct, pRO1887, contained 9.2 kilobases of PAO1 chromosomal DNA inserted in the PstI site of the vector pRO1727. Plasmid pRO1887 complemented only the tynC1 mutation. Mapping experiments performed with the chromosome-mobilizing plasmid R68.45 located the mutations described above in a cluster at about 35 to 40 min of the PAO1 chromosome map. The mutations were linked to the proA, thr-48, lys-9015, argF10, and argG markers.
mesh_terms: D015102:3,4-Dihydroxyphenylacetic Acid / Q000378:metabolism*; D001679:Biogenic Amines / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D009154:Mutation; D010648:Phenylacetates / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D001679:Biogenic Amines; D010648:Phenylacetates; D015102:3,4-Dihydroxyphenylacetic Acid