AN ANOMALOUS TRICARBOXYLIC ACID CYCLE IN ACETOBACTER MELANOGENUM

1960 ◽  
Vol 38 (3) ◽  
pp. 193-203 ◽  
Author(s):  
D. H. Bone ◽  
R. M. Hochster
Keyword(s):  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Oxidase System ◽  
Isocitric Dehydrogenase ◽  
Highly Active ◽  
Carbon Compounds ◽  
Condensing Enzyme

Extracts of gluconate-grown Acetobacter melanogenum contain condensing enzyme and DPN-isocitric dehydrogenase of low specific activity. No evidence could be found for the presence of phosphotransacetylase, aconitase, or TPN-isocitric dehydrogenase. Since the organism or its extracts cannot synthesize the necessary four carbon compounds from pyruvate and from acetate, it is concluded that the tricarboxylic acid cycle does not function in extracts of this organism in the usually accepted manner.The pyruvic oxidase system was found to be highly active, acetaldehyde being the chief intermediate and acetate the end product. The mechanism for the slow incorporation of acetate into other cell constituents is, at present, unknown.

AN ANOMALOUS TRICARBOXYLIC ACID CYCLE IN ACETOBACTER MELANOGENUM

1960 ◽  
Vol 38 (1) ◽  
pp. 193-203 ◽  
Author(s):  
D. H. Bone ◽  
R. M. Hochster
Keyword(s):  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Oxidase System ◽  
Isocitric Dehydrogenase ◽  
Highly Active ◽  
Carbon Compounds ◽  
Condensing Enzyme

Extracts of gluconate-grown Acetobacter melanogenum contain condensing enzyme and DPN-isocitric dehydrogenase of low specific activity. No evidence could be found for the presence of phosphotransacetylase, aconitase, or TPN-isocitric dehydrogenase. Since the organism or its extracts cannot synthesize the necessary four carbon compounds from pyruvate and from acetate, it is concluded that the tricarboxylic acid cycle does not function in extracts of this organism in the usually accepted manner.The pyruvic oxidase system was found to be highly active, acetaldehyde being the chief intermediate and acetate the end product. The mechanism for the slow incorporation of acetate into other cell constituents is, at present, unknown.

scholarly journals IN VITRO EFFECTS OF FLUORIDE ON TRICARBOXYLIC ACID CYCLE DEHYDROGENASES AND OXIDATIVE PHOSPHORYLATION: PART I

1967 ◽  
Vol 15 (4) ◽  
pp. 195-201 ◽  
Author(s):  
C. JAMES LOVELACE ◽  
GENE W. MILLER
Keyword(s):  
Oxidative Phosphorylation ◽  
Competitive Inhibition ◽  
Tricarboxylic Acid Cycle ◽  
Succinic Dehydrogenase ◽  
Tricarboxylic Acid ◽  
Initial Increase ◽  
Acid Cycle ◽  
Oxidase System ◽  
Vitro Effect

Studies were conducted on the in vitro effect of fluoride on the succinic oxidase system utilizing mitochondria obtained from cauliflower. Preincubation of mitochondria with fluoride did not increase inhibition of succinic oxidase. Various other tricarboxylic acid cycle substrates were used to determine their sensitivity to fluoride; only succinate oxidation was affected. A series of succinate concentrations in the presence and in the absence of fluoride showed increased activity of succinic dehydrogenase, which indicated competitive inhibition. Various concentrations of phosphate in the absence of fluoride showed that phosphate had only slight effects on the succinic 2,6-dichlorophenolindophenol reductase component of the succinic oxidase system. In the absence of phosphate, various concentrations of fluoride showed an initial increase in activity followed by a decrease in activity of succinic 2,6-dichlorophenolindophenol reductase. In the presence of phosphate, fluoride caused marked inhibition of succinic 2,6-dichlorophenolindophenol reductase. It is believed that this inhibition results from an enzyme-fluorophosphate complex which has a lower dissociation constant than that of the enzyme-substrate complex. An oxidative phosphorylation study indicated that both respiration and phosphorylation were inhibited.

METABOLISM OF C14 AMINO ACIDS AND AMIDES IN DETACHED LEAVES

1956 ◽  
Vol 34 (4) ◽  
pp. 423-433 ◽  
Author(s):  
C. D. Nelson ◽  
G. Krotkov
Keyword(s):  
Amino Acids ◽  
Double Bond ◽  
Glutamic Acid ◽  
Broad Bean ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Activity Data ◽  
Acid Cycle ◽  
Bean Leaves

Detached broad bean leaves were placed with their petioles in 0.01 M ammonium nitrate and allowed to carry on photosynthesis in C14O2 for various periods from 12 to 125 min. The radioactivities of the various amino acids formed from C14O2 were determined. In addition, these amino acids were degraded by decarboxylation with ninhydrin. From the specific activity data it was concluded that the amino acid closest to the site of carbon dioxide fixation in photosynthesis was alanine, followed by aspartic and glutamic acids, with the amides farthest removed. From the intramolecular distribution of label it was concluded that asparagine and glutamine were formed from their corresponding amino acids. The labelling in aspartic and glutamic acids was not consistent with the view that these two amino acids are formed from their corresponding α-keto acids produced by operation of the conventional tricarboxylic acid cycle. A C2 plus C2 condensation is postulated for the formation of aspartic acid. A shift in the double bond in the aconitase reaction of the tricarboxylic acid cycle would account for the observed labelling in glutamic acid. When acetate-1-C14 was fed to detached broad bean leaves in the light or dark, the distribution of label in glutamic acid supported the suggestion that there is such a. shift in the double bond in the aconitase reaction. Sodium arsenite, infiltrated into tobacco leaves, inhibited the biosynthesis of asparagine but not that of glutamine.

THE TRICARBOXYLIC ACID CYCLE, THE GLYOXYLATE CYCLE, AND THE ENZYMES OF GLUCOSE OXIDATION IN PSEUDOMONAS AERUGINOSA

1966 ◽  
Vol 12 (5) ◽  
pp. 1015-1022 ◽  
Author(s):  
Margaret von Tigerstrom ◽  
J. J. R. Campbell
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Glucose Oxidation ◽  
Nadh Oxidase ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Glyoxylate Cycle ◽  
Succinic Dehydrogenase ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
The Glyoxylate Cycle

The enzymes of the glyoxylate cycle, the tricarboxylic acid cycle, glucose oxidation, and hydrogen transport were measured in extracts of Pseudomonas aeruginosa grown with glucose, α-ketoglutarate, or acetate as sole carbon source. The specific activity of isocitritase was increased 25-fold by growth on acetate whereas malate synthetase was increased only 4-fold. All of the enzymes of glucose metabolism, operative at the hexose level, were inducible. The enzymes of the tricarboxylic acid cycle were present under all conditions of growth but extracts from acetate-grown cells contained only one-quarter of the fumarase and pyruvic oxidase activity and half the malate-oxidizing activity of the other extracts. Transhydrogenase, NADH oxidase, and NADPH oxidase activities were similar in each type of extracts. Most of the enzymes were present in the soluble cytoplasm, exceptions being glucose oxidase, succinic dehydrogenase, and NADH oxidase.

Studies on wheat plants using 14C-labelled compounds. XXII. Incorporation into wheat protein

1969 ◽  
Vol 47 (1) ◽  
pp. 19-23 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Glutamic Acid ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Radioactive Tracers ◽  
Wheat Protein ◽  
Acid Cycle ◽  
Late Stages ◽  
Kernel Protein

Thatcher wheat plants were labelled with 14C by injecting radioactive tracers into the top internode of the stem during late stages of plant growth. The distribution of 14C in fully mature plants was examined, emphasis being placed on the labelling of kernel-protein amino acids.Glutamine was only slightly more effective than glutamic acid for labelling glutamic acid isolated from the gluten hydrolysate, indicating that glutamic acid and glutamine are extensively interconverted in the wheat plants. Proline and glutamic acid also are readily interconverted, proline-14C yielding protein in which the glutamic acid has a higher specific activity than does the proline. By contrast, arginine-5-14C did not yield highly labelled glutamic acid.14C from glyoxylate-1-14C was widely distributed among kernel components but it produced glycine and serine with carboxyl carbons of exceptionally high specific activity.Succinate-1,4-14C, succinate-2,3-14C, and aspartic acid-14C all labelled glutamic acid of kernel protein more extensively than the other amino acids of the protein. The role of the tricarboxylic acid cycle in utilizing these tracers is discussed.

scholarly journals The Prevalence of Carbon-13 in Respiratory Carbon Dioxide As an Indicator of the Type of Endogenous Substrate

1970 ◽  
Vol 55 (1) ◽  
pp. 1-17 ◽  
Author(s):  
Bruce S. Jacobson ◽  
Bruce N. Smith ◽  
Samuel Epstein ◽  
George G. Laties
Keyword(s):  
Isotope Ratio ◽  
Tricarboxylic Acid Cycle ◽  
Carbon Isotope Ratio ◽  
Tricarboxylic Acid ◽  
Long Chain Fatty Acids ◽  
Tuber Tissue ◽  
Acid Cycle ◽  
Carbon Compounds ◽  
Endogenous Substrate ◽  
Long Chain

Isotope discrimination is a common feature of biosynthesis in nature, with the result that different classes of carbon compounds frequently display different 13C/12C ratios. The 13C/12C ratio of lipid in potato tuber tissue is considerably lower than that for starch or protein. We have collected respiratory CO2 from potato discs in successive periods through 24 hr from the time of cutting—an interval in which the respiration rate rises 3–5-fold. The 13C/12C ratio of the evolved CO2 was determined for each period, and compared with the 13C/12C ratios of the major tissue metabolites. In the first hours the carbon isotope ratio of the CO2 matches that of lipid. With time, the ratio approaches that typical of starch or protein. An estimation has been made of the contribution of lipid and carbohydrate to the total respiration at each juncture. In connection with additional observations, it was deduced that the basal, or initial, respiration represents lipid metabolism—possibly the α-oxidation of long chain fatty acids—while the developed repiration represents conventional tricarboxylic acid cycle oxidation of the products of carbohydrate glycolysis. The true isotopic composition of the respiratory CO2 may be obscured by fractionation attending the refixation of CO2 during respiration, and by CO2 arising from dissolved CO2 and bicarbonate preexisting in the tuber. Means are described for coping with both pitfalls.

scholarly journals The pyruvate-oxidase system in brain and the tricarboxylic acid cycle*

1949 ◽  
Vol 45 (3) ◽  
pp. 320-325 ◽  
Author(s):  
R. V. Coxon ◽  
C. Liébecq ◽  
R. A. Peters
Keyword(s):  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Pyruvate Oxidase ◽  
Acid Cycle ◽  
Oxidase System

Use of 14CO2 in estimating rates of hepatic gluconeogenesis

1992 ◽  
Vol 263 (1) ◽  
pp. E36-E41 ◽  
Author(s):  
E. Esenmo ◽  
V. Chandramouli ◽  
W. C. Schumann ◽  
K. Kumaran ◽  
J. Wahren ◽  
...  
Keyword(s):  
Blood Glucose ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Hepatic Gluconeogenesis ◽  
Acid Cycle ◽  
Specific Activities ◽  
Glucose Formation

Estimating the rate of hepatic gluconeogenesis in vivo from the incorporation of 14C from 14CO2 into glucose requires determination of the rates in liver of equilibration of oxaloacetate with fumarate, conversion of oxaloacetate to phosphoenolpyruvate (PEP), and conversion of PEP to pyruvate, all relative to the rate of tricarboxylic acid cycle flux. With the use of a model of mitochondrial metabolism and gluconeogenesis, expressions are derived relating specific activity of carboxyl of PEP from 14CO2 to those rates and specific activity of mitochondrial CO2. If those rates and specific activity of mitochondrial CO2 are known, specific activity of PEP, calculated using the expressions, should, on a mole basis, be one-half the specific activity of the glucose formed. At steady state, in the 60-h fasted individual, where glucose formation is solely by gluconeogenesis, twice estimated specific activity of PEP should then approximate that of blood glucose. Estimates of relative rates in 60-h fasted humans, previously made from distribution of 14C in glutamate from phenylacetylglutamine excreted when [3-14C]lactate and phenylacetate were given, were applied to the expressions. Specific activity of mitochondrial CO2 was equated to that of CO2 expired by 60-h fasted subjects given NaH14CO3 and alpha-[1-14C]ketoisocaproate. Predicted specific activities approximated actual specific activities of blood glucose when NaH14CO3 was administered. alpha-[1-14C]ketoisocaproate administrations gave underestimates. This is attributable to differences between specific activities of hepatic mitochondrial CO2 and expired CO2, which is evidenced by higher incorporations of 14C in glucose than in expired CO2 from alpha-[1-14C]ketoisocaproate than from NaH14CO3.(ABSTRACT TRUNCATED AT 250 WORDS

Acetate metabolism in bovine ketosis

1959 ◽  
Vol 14 (6) ◽  
pp. 1029-1032 ◽  
Author(s):  
D. S. Kronfeld ◽  
Max Kleiber ◽  
J. M. Lucas
Keyword(s):  
Carbon Dioxide ◽  
Plasma Glucose ◽  
Milk Fat ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acetate Metabolism ◽  
Acid Cycle ◽  
The Mean ◽  
Fat Synthesis

The metabolism of intravenously injected acetate–1–C14 was compared in normal and ketotic cows. The mean standardized specific activity of milk citrate, casein, lactose, plasma glucose, respired carbon dioxide and urine acetone was greater in the ketotic than in the normal cows, while that of milk fat was markedly decreased. The total–C14 recovered in the milk fat within 48 hours was about 14% of the injected dose in the normal cows, but only about 3% in the ketotic cows. These results suggest that the proportion of acetate metabolized via the tricarboxylic acid cycle is increased, while milk fat synthesis from acetate is impaired during bovine ketosis. Submitted on December 15, 1958

STUDIES ON WHEAT PLANTS USING C14 COMPOUNDS: V. GERMINATION STUDIES WITH LABELLED WHEAT SEEDS

1957 ◽  
Vol 35 (1) ◽  
pp. 1259-1266 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Carbon Dioxide ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Active Compound ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Carbon 14 ◽  
Wheat Seeds

Radioactive wheat seeds, obtained by injecting acetate-C14 into the stems of the parent plants, were germinated in the absence of light and nutrient and the fate of the carbon-14 was observed. Carbon respired as carbon dioxide had a higher specific activity than any of the major seed components except protein. Variations were found in the patterns by which material was transferred from the kernel to new tissue as reflected in a comparison of the activity of various components. Glutamic acid was the most active compound isolated either from the original seeds or from the new tissues. This observation, together with similarities noted in the intramolecular distribution of carbon-14 in glutamic acid of new tissue and seed residues, indicated that glutamic acid was reutilized for the biosynthesis of seedling protein. Changes in the labelling of glutamic acid during transfer to new tissue are qualitatively in accord with the idea that at least some of the amino acid is used after re-entry into the tricarboxylic acid cycle.

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