Oxidation of Amino Acids and Compounds Associated With the Tricarboxylic Acid Cycle by Neisseria Gonorrhoeae

Tonhazy, Nicholas E.; Pelczar, Michael J.

doi:10.1128/jb.65.4.368-377.1953

Cited by 14 publications

(4 citation statements)

References 22 publications

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“…Results somewhat similar to ours were obtained by Tonhazy and Pelczar (8) with N. gonorrhoeae. Respiration was elicited by glutamate only after a prolonged lag period.…”

Section: Andsupporting

confidence: 92%

Catabolic Activities of Neisseria meningitidis: Utilization of Glutamate

Mallavia

Weiss

1970

J Bacteriol

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Glutamate was catabolized at a rapid rate by Neisseria meningitidis, group B. Surprisingly, there was a lag of 5 to 30 min in respiration, but not in CO2 production from C1, and an appreciable amount of succinate accumulated. The eventual rapid rate of respiration was not prevented by the addition of chloramphenicol. The lag period was eliminated by combinations of substrates that favored the activity of a glutamate-oxaloacetate transaminase. It is suggested that with glutamate as the sole substrate, the reaction terminated at succinate, required only moderate 02 uptake, and did not result in the transport of succinate to enzymatic sites. The lag period represented the time required for the accumulation of succinate and its transport to enzymatic sites by energy provided by the metabolism of the remaining glutamate. When the transaminase was operative, on the other hand, successive products of the reaction were immediately placed in contact with enzymatic sites.During the course of experiments designed to characterize, by simple metabolic tests, strains of Neisseria meningitidis of different origins, it was noted that greatest variation occurred in the utilization of glutamate and a-ketoglutarate. The respiration of one of the strains, M 2091, group B, was greatly stimulated by glutamate, but only after a lag period, which was apparently due to a temporary block of succinate utilization. MATERIALS AND METHODSThe strain M 2091, group B, of N. meningitidis was obtained from the collection of Sara E. Branham, who in turn received it from the Massachusetts Department of Health on 29 October 1947 as a nasal isolate (1). In our laboratory, the microorganism was passed twice and seed cultures were maintained at -70 C.For each experiment, the organisms were cultivated for 24 hr on Mueller-Hinton agar slants at 37 C, the growth was suspended in 5 ml of Trypticase Soy Broth (TSB), and a 0.3-ml amount of the suspension was inoculated into 100 ml of TSB. Cultures inoculated in this manner and incubated at 32 C for 16 to 18 hr were near the end of the period of exponential growth. The cells were washed once and suspended in 0.1 M phosphate buffer (disodium and monopotassium), pH 7.4. The cell density was adjusted spectrophotometrically to 0.7 to 1.5 mg of cell protein or 1.5 X 109 to 7.5 X 109 viable cells per ml.Oxygen uptake was measured by standard manometric techniques. CO2 production was determined by . 99163.using '4C-labeled substrates in 25-ml flasks fitted with plastic cups containing Hyamine (Packard Instrument Co., La Grange, Ill.) to trap the radioactive CO2 as described by Weiss (9). In some cases, the two determinations were combined by using labeled substrates in Warburg flasks and trapping the CO2 in 0.2 ml of 10 N KOH placed in the center well. At the end of the incubation period, 0.2 ml of concentrated sulfuric acid or citric acid was tipped into the reaction mixture, and the trapping of CO2 was continued for an additional 0.5 hr. Unless otherwise indicated, the flasks contained 1.0 ml of the cell suspension...

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“…Results somewhat similar to ours were obtained by Tonhazy and Pelczar (8) with N. gonorrhoeae. Respiration was elicited by glutamate only after a prolonged lag period.…”

Section: Andsupporting

confidence: 92%

Catabolic Activities of Neisseria meningitidis: Utilization of Glutamate

Mallavia

Weiss

1970

J Bacteriol

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“…This pattern of acetate oxidation is indicative of a functioning tricarboxylic cycle. These results were contrary to those of Tonhazy and Pelczar (41) who utilized resting-cell suspensions ofglucose-grown N. gonorrhoeae and concluded that of the members of the conventional tricarboxylic acid cycle. only a-ketoglutarate, succinate, fumarate, malate, and oxaloacetate could be oxidized.…”

contrasting

confidence: 99%

“…In spite of a functional tricarboxylic acid cycle, the gonococcus cannot grow in medium supplemented with tricarboxylic acid cycle intermediates. The lack of permeability does not appear to be a factor (20,41). An 6.…”

Section: Discussionmentioning

confidence: 99%

Physiology and metabolism of pathogenic neisseria: tricarboxylic acid cycle activity in Neisseria gonorrhoeae

Hebeler¹,

Morse²

1976

J Bacteriol

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Tricarboyxlic acid cycle activity was examined in Neisseria gonorrhoeae CS-7. The catabolism of glucose in N. gonorrheae by a combination of the Entner-Doudoroff and pentose phosphate pathways resulted in the accumulation of acetate, which was not further catabolized until the glucose was depleted or growth became limiting. Radiorespirometric studies revealed that the label in the 1 position of acetate was converted to CO2 at twice the rate of the label in the 2 position, indicating the presence of a tricarboxylic acid cycle. Growth on glucose markedly reduced the levels of all tricarboxylic acid cycle enzymes except citrate synthase (EC 4.1.3.7). Extracts of glucose-grown cells contained detectable levels of all tricarboxylic acid cycle enzymes except aconitase (EC 4.2.1.3), isocitrate dehydrogenase (EC 1.1.1.42), and a pyridine nucleotide-dependent malate dehydrogenase (EC 1.1.1.37). Extracts of cells capable of oxidizing acetate lacked only the pyridine nucleotide-dependent malate dehydrogenase. In lieu of this enzyem, a particulate pyridine nucleotide-independent malate oxidase (EC 1.1.3.3) was present. This enzyme required flavin adenine dinucleotide for activity and appeared to be associated with the electron transport chain. Radiorespirometric studies utilizing labeled glutamate demonstrated that a portion of the tricarboxylic acid cycle functioned during glucose catabolism. In spite of the presence of all tricarboxylic acid cycle enzymes, N. gonorrhoeae CS-7 was unable to grow in medium supplemented with cycle intermediates.

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“…Aspartase (EC 4.3.1.1) can also catalyze the de novo synthesis of a-amino groups, but this enzyme is generally repressed during cultivation in minimal media indicating that it is primarily associated with the catabolism of aspartate (13,34). Glutamate dehydrogenase can also function as a catabolic enzyme by catalyzing the oxidative deamination of glutamate, a reaction which serves as an important source of energy for a number of obligate and facultative intracellular bacterial parasites (2,5,19,21,22,29,31).…”

mentioning

confidence: 99%

Mutations Influencing the Assimilation of Nitrogen by Yersinia pestis

Brubaker

Sulen

1971

Infect Immun

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Cells of 20 isolates of Yersinia (Pasteurella) pestis exhibited an unusual nutritional requirement which could be fulfilled by glycine or L-threonine. Meiotrophic mutants which required neither of these amino acids (Gly/Thr+) were isolated from cultures of all 20 strains at a frequency of 10-7. Wild-type and Gly/Thr+ cells of 14 strains failed to utilize L-amino acids or urea (0.01 M) as primary sources of nitrogen and grew slowly in the presence of low concentrations of NH4+ (< 5 mM). Cells of six strains (termed N+) utilized certain L-amino acids and urea (0.01 M) as primary sources of nitrogen and grew rapidly in the presence of 5 mM NH4+. N+ but not Norganisms cultivated with NH4+ (0.01 M) as a primary source of nitrogen excreted a complete spectrum of naturally occurring amino acids; under this condition of growth the aspartase and particulate nicotinamide adenine dinucleotide phosphate transhydrogenase activities of N+ and N-cells were repressed. N+ meiotrophs arose at a frequency of 10t-in cultures of all 14 Nisolates, and urease-positive meiotrophs could be selected at a frequency of 10-7 from N+ but not N-cells of all 20 strains on a medium containing urea (0.01 M) as a primary source of nitrogen. These findings illustrate a reversible loss of genetic potential which has occurred during the evolution of Y. pestis as an obligate parasite and suggest that this organism is unable to efficiently remove dispensable deoxyribonucleic acid from its chromosome.

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Oxidation of Amino Acids and Compounds Associated With the Tricarboxylic Acid Cycle by Neisseria Gonorrhoeae

Cited by 14 publications

References 22 publications

Catabolic Activities of Neisseria meningitidis: Utilization of Glutamate

Catabolic Activities of Neisseria meningitidis: Utilization of Glutamate

Physiology and metabolism of pathogenic neisseria: tricarboxylic acid cycle activity in Neisseria gonorrhoeae

Mutations Influencing the Assimilation of Nitrogen by Yersinia pestis

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