Formation and operation of the histidine-degrading pathway in Pseudomonas aeruginosa.
Histidine ammonia lyase (histidase), urocanase, and the capacity to degrade formiminoglutamate, which are respectively involved in steps I, II, and IV in the catabolism of histidine, were induced during growth of Pseudomonas aeruginosa on histidine or urocanate, and were formed gratuitously in the presence of dihydro-urocanate. Urocanase-deficient bacteria formed enzymes I and IV constitutively; presumably they accumulate enough urocanate from the breakdown of endogenous histidine to induce formation of the pathway. Urocanate did not satisfy the histidine requirement of a histidine auxotroph, indicating that it probably acted as an inducer without being converted to histidine. The results imply that urocanate is the physiological inducer of the histidine-degrading enzymes in P. aeruginosa. Enzymes of the pathway were extremely sensitive to catabolite repression; enzymes I and II, but not IV, were coordinately repressed. Our results suggest a specific involvement of nitrogenous metabolites in the repression. Mutant bacteria with altered sensitivity to repression were obtained. The molecular weight of partially purified histidase was estimated at 210,000 by sucrose gradient centrifugation. Its K(m) for histidine was 2 x 10(-3)m in tris(hydroxymethyl)aminomethane chloride buffer. Sigmoid saturation curves were obtained in pyrophosphate buffer, indicating that the enzyme might have multiple binding sites for histidine. Under certain conditions, histidase appeared to be partially inactive in vivo. These findings suggest that some sort of allosteric interaction involving histidase may play a role in governing the operation of the pathway of histidine catabolism.
D000581:Amidohydrolases / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003470:Culture Media; D011756:Diphosphates / Q000494:pharmacology; D004790:Enzyme Induction; D004794:Enzyme Repression; D005365:FIGLU Test; D006639:Histidine / Q000378:metabolism*; D008190:Lyases / Q000032:analysis / Q000378:metabolism*; D009154:Mutation; D010101:Oxygen Consumption; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000378:metabolism*
D003470:Culture Media; D011756:Diphosphates; D006639:Histidine; D000581:Amidohydrolases; D008190:Lyases