Occurrence of succinyl derivatives in the catabolism of arginine in Pseudomonas cepacia (original) (raw)
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N2-Succinylated Intermediates in an Arginine Catabolic Pathway of Pseudomonas aeruginosa
Proceedings of The National Academy of Sciences, 1986
Arginine-nonutilizing (aru) mutants of Pseudomonas aeruginosa strain PAO converted L-arginine to N2succinylarginine or N-succinylglutamate, which were identified by high-voltage electrophoresis and HPLC. Addition of aminooxyacetate, an inhibitor of pyridoxal phosphate-dependent enzymes, to resting cells of the wild-type PAO1 in arginine medium led to the accumulation of N2-succinylornithine. Enzyme assays with crude P. aeruginosa extracts established the
European Journal of Biochemistry, 1994
The arginine and ornithine succinyltransferase from Pseudomonas aeruginosa, a bifunctional enzyme involved in the aerobic utilization of arginine and ornithine, has been purified to homogeneity. The apparent molecular mass of the native enzyme was 150 kDa by gel filtration and 140 kDa by polyacrylamide gel electrophoresis under non-denaturing conditions. After SDSPAGE two subunits of 35 kDa and 37 kDa were evident, indicating that the enzyme is a heterotetramer. Microsequence analysis of the electroblotted protein bands gave two different but well-conserved N-terminal amino acid sequences.
Catabolism of Arginine, Citrulline and Ornithine by Pseudomonas and Related Bacteria
Microbiology-sgm, 1987
The distribution of the arginine succinyltransferase pathway was examined in representative strains of Pseudomonas and related bacteria able to use arginine as the sole carbon and nitrogen source for growth. The arginine succinyltransferase pathway was induced in arginine-grown cells. The accumulation of succinylornithine following in vivo inhibition of succinylornithine transaminase activity by aminooxyacetic acid showed that this pathway is responsible for the dissimilation of the carbon skeleton of arginine. Catabolism of citrulline as a carbon source was restricted to relatively few of the organisms tested. In P. putida, P . cepacia and P . indigofera, ornithine was the main product of citrulline degradation. In most strains which possessed the arginine succinyltransferase pathway, the first step of ornithine utilization as a carbon source was the conversion of ornithine into succinylornithine through an ornithine succinyltransferase. However P . cepacia and P. putida used ornithine by a pathway which proceeded via proline as an intermediate and involved an ornithine cyclase activity.
L-Arginine Utilization by Pseudomonas Species
Microbiology, 1984
The utilization of arginine was studied in several different Pseudomonas species. The arginine decarboxylase and agmatine deiminase pathways were found to be characteristic of Pseudomonas species of group I as defined by Palleroni et al. (1 974). Pseudomonas putida strains had three distinct arginine catabolic pathways initiated by arginine decarboxylase, arginine deiminase and arginine oxidase, respectively. The two former routes were also present in P. Jluorescens and P. mendocina and in P. aeruginosa which also used arginine by a further unknown pathway. None of these pathways occurred in P. cepacia strains; agmatine catabolism seemed to follow an unusual route involving guanidinobutyrate as intermediate. Chemicals. Arginine, ornithine, citrulline, putrescine and guanidinobutyrate, and L-amino acid oxidase from Crotalus adamenreus, were obtained from Sigma. DL[ l-'JC]Arginine. HCl was purchased from CEA-IRE, Fleurus (Belgium). Agmatine was obtained from Aldrich and found to be free from putrescine. N-Carbamoylputrescine and N-[carbamoy/-'4C]carbamoylputrescine were prepared as previously described (Mercenier et a/., 1 9 8 0~). 2-Ketoarginine was synthesized according to Meister (1952, 1953). Arginine was oxidized enzymically by L-amino acid oxidase for 12 h, after which the solution was filtered using a Diaflo apparatus, the enzyme recovered, and the filtrate was concentrated under reduced pressure, 2-Ketoarginine was crystallized at 4 "C and the precipitate redissolved in hot water. Residual arginine was separated from 2-ketoarginine by chromatography on Dowex 50 WX8 (H + form). The column (2.5 x 30 cm) was washed with 1 I water, which eluted the 2-ketoarginine, the arginine remaining adsorbed. The eluate was concentrated and 2-ketoarginine was recrystallized at 4 "C. A 5% (w/v) solution was found essentially free of arginine as determined by amino acid analysis (Mercenier et al.,
Regulation of enzyme synthesis in the arginine biosynthetic pathway of Pseudomonas aeruginosa
Journal of general microbiology, 1978
In Pseudomonas aeruginosa the synthesis of only two out of eight arginine biosynthetic enzymes tested was regulated. Comparisons were made between the specific activities of these enzymes in bacteria grown on arginine or on its precursor, glutamate. N2-Acetylornithine 5-aminotransferase (ACOAT), an enzyme involved in both the biosynthesis and catabolism of arginine, was induced about 14-fold during growth of the organism on arginine as the only carbon and nitrogen source, and the anabolic ornithine carbamoyltransferase (aOTC), a strictly biosynthetic enzyme, was repressed 18-fold. Addition of various carbon sources to the arginine medium led to repression of ACOAT and to derepression of aOTC. Fructose, which supported only slow growth of P. aeruginosa, had a weak regulatory effect on the synthesis of the two arginine enzymes while citrate, a good carbon source for this organism, had a strong effect. The repression of ACOAT by citrate was not relieved by adding cyclic AMP to the medi...
N 2 -Succinylornithine in ornithine catabolism of Pseudomonas aeruginosa
Archives of Microbiology, 1988
Most Pseudomonas aeruginosa PAO mutants which were unable to utilize l-arginine as the sole carbon and nitrogen source (aru mutants) under aerobic conditions were also affected in l-ornithine utilization. These aru mutants were impaired in one or several enzymes involved in the conversion of N2-succinylornithine to glutamate and succinate, indicating that the latter steps of the arginine succinyltransferase pathway can be used for ornithine catabolism. Addition of aminooxyacetate, an inhibitor of the N2-succinylornithine 5-aminotransferase, to resting cells of P. aeruginosa in ornithine medium led to the accumulation of N2-succinylornithine. In crude extracts of P. aeruginosa an ornithine succinyltransferase (l-ornithine:succinyl-CoA N2-succinyltransferase) activity could be detected. An aru mutant having reduced arginine succinyltransferase activity also had correspondingly low levels of ornithine succinyltransferase. Thus, in P. aeruginosa, these two activities might be due to the same enzyme, which initiates aerobic arginine and ornithine catabolism.
Journal of bacteriology, 1975
In Pseudomonas aeruginosa N-2-acetylornithine 5-aminotransferase (ACOAT), the fourth enzyme of arginine biosynthesis is induced about 15-fold by cultivating the organism on a medium with L-arginine as the sole carbon and nitrogen source. Synthesis of the enzyme is subject to catabolite repression and nitrogen source. Synthesis of the enzyme is subject to catabolite repression by a variety of carbon sources. ACOAT from strain PAO 1 was purified over 40-fold to electrophoretic homogeneity. A molecular weight of approximately 110,000 was obtained by thin-layer gel filtration. Electrophoresis in sodium dodecyl sulfate gels gave a single band corresponding to a molecular weight of 55,000. Purified ACOAT catalyzes the transamination of N-2-acetyl-L-ornithine as well as of L-ornithine with 2-oxoglutarate (Km values of 1.1, 10.0, and 0.7 mM, respectively). With N-2-acetyl-L-ornithine as amino donor, the pH-optimum of the enzymatic reaction is 8.5; with L-ornithine as amino donor, 9.5. The c...
Regulation of enzyme synthesis in the arginine deiminase pathway of Pseudomonas aeruginosa
Journal of Bacteriology, 1980
The three enzymes of the arginine deiminase pathway in Pseudomonas aeruginosa strain PAO were induced strongly (50- to 100-fold) by a shift from aerobic growth conditions to very low oxygen tension. Arginine in the culture medium was not essential for induction, but increased the maximum enzyme levels twofold. The induction of the three enzymes arginine deiminase (EC 3.5.3.6), catabolic ornithine carbamoyltransferase (EC 2.1.3.3), and carbamate kinase (EC 2.7.2.3) appeared to be coordinate. Catabolic ornithine carbamoyltransferase was studied in most detail. Nitrate and nitrite, which can replace oxygen as terminal electron acceptors in P. aeruginosa, partially prevented enzyme induction by low oxygen tension in the wild-type strain, but not in nar (nitrate reductase-negative) mutants. Glucose was found to exert catabolite repression of the deiminase pathway. Generally, conditions of stress, such as depletion of the carbon and energy source or the phosphate source, resulted in induc...