Oxalacetate transamination

The procedure reported in Scheme 13.11 describes deracemization of an amino acid involving oxidation with an L-specific enzyme and transamination with a D-amino transferase using D-aspartate 10, which is generated from L-aspartate 11 by aspartate racemase, as the amino donor. The oxidative enzyme is defined as an L-amino acid deaminase, a flavoprotein from Proteus myxofadens [34]. The transamination reaction is shifted towards the product since the oxalacetate 12 formed decarboxylates spontaneously to give pyruvate and carbon dioxide. 

In the hydrogenolytic asymmetric transamination between a benzylic amine and oxalacetic acid the Schiff s base was hydrogenated in ethanol over palladium, but alanine rather than aspartic acid was the... 

It is apparent that methodological problems limit the usefulness of the kinetic approach to studying mechanisms. However, data of an entirely different nature argue against a sequential model for the aspartate transporter. Studies of the metabolism of cysteine sulfinic acid show that it is transported by the glutamate/aspartate transporter and that it transaminates with a-ketoglutarate or oxalacetate to yield glutamate or aspartate and jS-sulfinyl pyruvate, which spontaneously hydrolyzes into sulfite and pyruvate [148,149]. 

The urea cycle is the most important process in biological ammonia detoxication, (s. pp 57, 266) (s. figs. 3.12, 3.13) It is directly linked with amino-acid metabolism and thus also with NH2 donors and precursors through specific amino acids and transamination processes. Here, the major transamination processes are those involving glutamate and oxalacetate as well as a-ketoglutarate and aspartate. 

C Amino acid s Asn is hydrolyzed in one step to aspaartate, which in turn is transaminated in one step to oxalacetate. Threonine feeds into the TCA cycle through succinyl-CoA instead of oxalacetate. Thr is first deaminated via a dehydratase as seen earlier, then decarboxylated by Pyruvate DH Complex to give propionyl-CoA, which is then transformed via a series of steps to give succinyl-CoA. 

Kumagai and coworkers11131 developed an enzymatic procedure to produce d-alanine from fumarate by means of aspartase (E. C. 4.3.1.1), aspartate racemase, and D-amino acid aminotransferase (Fig. 17-12). Aspartase catalyzes conversion of fumarate into L-aspartate, which is racemized to form D-aspartate. D-Amino acid aminotransferase catalyzes transamination between D-aspartate and pyruvate to produce D-alanine and oxalacetate. This 2-oxo acid is easily decarboxylated spontaneously to form pyruvate in the presence of metals. Thus, the transamination proceeds exclusively toward the direction of D-alanine synthesis, and total conversion of fumarate into D-alanine was achieved. 

Deamination of amino acids in animal tissue is generally effected by transamination with an a-keto-acid. In the majority of cases, this is 2-oxoglutarate formed by the citric acid cycle. Aspartate aminotransferase and alanine aminotransferase are examples of this kind of reaction. In Figure 2.7, transamination involving these enzymes is depicted as it is known to occur in mammalian liver. Note that the scheme shown here requires participation of oxalacetate and pyruvate and thus is intimately connected with metabolic pathways considered earlier. Serine and glycine are readily interconvertible in animal tissue by the enzyme serine hydroxymethyltransferase. It is worth noting also that decarboxylation of serine to ethanolamine as mentioned above can be followed by A -methylation to yield choline. Choline is both an essential component of many... 

In a similar way, transamination from aspartate (Asp, D) to a-ketoglutarate, which, like fumarate, is produced in the tricarboxylic acid cycle (Scheme 11.89), then yields glutamate (Glu, E) and, exactly as in Scheme 12.2, oxalacetate (Equation 12.6). 

The basic construction of the mathematical model using simplified metabolic networks to describe the reactions of the citric acid cycle and associated transamination reactions between pyruvate and alanine, oxalacetate and aspartate and a-ketoglutarate and glutamate, and the use of the FACSIMILE program (Chance et al., 1977) to solve the rather large number of simultaneous differential equations generated by the model was the same as previously described (Chance etal., 1983). For the present experiments the model was expanded to include an input flux at the level of succinate to represent propionate metabolism to succinyl-CoA, and a dilution of the aspartate pool to represent net proteolysis. These input fluxes required an output flux of carbon from the citric acid cycle in order to maintain a steady state carbon balance, for which the conversion of malate to pyruvate via malic enzyme was chosen. The model calculates the unknown flux parameters to provide a minimum least squares fit of the C fractional enrichments of specific carbon atoms of metabolic intermediates as measured by C NMR spectroscopy. 

Assay of Transamination. Since the sum of keto acids and amino acids does not change in a transamination, specific reactions are required to assay the reaction products. Some of the methods used are oxidation of a-ketoglutarate to succinate and determination of succinate with succinic dehydrogenase decarboxylation of oxalacetate with aniline citrate decarboxylation with specific amino acid decarboxylases separation of products on paper chromatograms and spectrophotometric determination of those keto acids that exhibit specific absorption. 

Sulfinylpyruvic acid accumulates as a result of the transaminating activity. It is an analog of oxalacetic acid, and like this compound, it is decomposed to pyruvate and SOs under the influence of Mn ". The reaction is analogous to the j3-decarboxylation of oxalacetate by Mn++. Mn++ also catalyzes the oxidation of SOj to S04 . As a consequence, reactions 5 and 6 of Fig. 2 are assumed to be nonenzymatic. 

The reported molecular weight is about 110,000 and the enzyme has 2 mol pyridoxal phosphate per mol of enzyme. It is reasonable to assume that the aspartate formed during CAM metabolism is by glutamate transamination to oxalacetate. 

The mechanism of transamination has been investigated by Jenkins and Sizer (498) vdio describe the preparation of distinct pyridoxal phosphate and pyridoxamine phosphate forms of glutamic-oxalacetic transaminase. Mason (499) has studied the inhibition of rat kidney kynurenine trans-... 

In the degradation of aspartic acid, the major pathway in the vertebrate organism appears to be its transamination to oxalacetic acid the latter is then oxidized by means of the TCA cycle. 

After weighing the different posibilities to counter the lack of equal labeling in carbons 3 and 6 of lysine, Strassman and Weinhouse 180) propose as the more likely synthetic mechanism, a condensation of an acetyl methyl carbon with the carbonyl carbon of a-ketoglutarate, similar to the condensation of acetyl CoA and oxalacetate to yield citrate. This reaction would form homocitric acid. Upon oxidation and decarboxylation of the latter there would be obtained a-ketoadipic acid. Transamination of a-ketoadipic acid produces a-aminoadipic acid, which can be converted to lysine by reduction to the corresponding semialdehyde, followed by transamination. 

A role for a-keto-c-aminopimelic acid on the pathway of lysine syntheds is indicated by the discovery of a transaminase that acts on both diaminopimelic acid and on lysine 194). Cell suspendons of acetone-dried bacteria at pH 5 were able to transaminate all three stereoisomers of diaminopimelic acid and also d- and L-lysine in the presence of pyridoxal phosphate, with pyruvate, oxalacetate, and a-ketoglutarate serving as amino group acceptors. 

Figures 4 and 5 illustrates the use of NMR spectroscopy to study the metabolism of (l-i C)glu-cose in primary cultures of neurons and astrocytes. A simplified scheme of the metabolism of (l-i C)glu-cose in neural cells is given in Figure 4. Briefly, (1-i C)glucose is metabolized to (3- C)pyruvate through the Embden Meyerhoff glycolytic pathway. The (3-i C)pyruvate produced can be transaminated to (3- C)alanine, reduced to (3-i C)lactate or enter the tricarboxylic acid (TCA) cycle through the pyruvate dehydrogenase (PDH) or pyruvate carboxylase (PC) activities. A net increase in (3-i C)lactate reveals increased aerobic glycolysis and is normally observed under hypoxic conditions in normal cells. If (3-i C)pyruvate enters the TCA cycle though PDH it produces (2-i C)acetyl-coenzyme A first, and subsequently (4-i C)a-ketoglutarate. In contrast, if (3-i C)pyruvate is carboxylated to (3-i C)oxalacetate by...

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