Citrate cycle carboxylic acids

The primary technology for citric acid recovery from fermentation broths is through precipitation with CaCOs, although other technologies include ion exchange, carbon adsorption, membrane filtration, chromatography, and liquid extraction [29]. Unlike other TCA cycle carboxylic acids, citric acid is excreted in the acid form rather than precipitated as citrate salt [30], which has the implication that the cell mass can first be filtered, followed by crystallization with iime with subsequent separation by filtration. The filter cake is next acidified with sulfuric acid, which precipitates the calcium as gypsum and the aqueous citric acid can next be decolourized and recrystallized by evaporation [29]. 

The formation of acetate CH3C02 + H+ from C02 and CH4. The acetyl group CH3CO- is the original building block of other carboxylic acids, by the reverse citrate cycle (Figure 4.4), and of lipids in cells. 

The addition of ammonia to the variety of acids derivable from either the breakdown of glucose, glycolysis, or of the pentose shunt reaction products, ribose and NADPH, and from the citrate cycle, gives the amino acids (see Table 4.7 and Figure 4.4) Polymerisation of amino acids in cells gives proteins. In some of the amino acids sulfur and selenium can be incorporated easily. We assume NH3 was present. (Note that Se is in a coded amino acid not in Table 4.7.) Some selective metal-binding properties can be seen in Table 4.7, but amino acid carboxylates can bind all. 

During the formation of carboxylic acid like lA, there will be shuttling of metabolites within the intracellular compartments, having the capability to utilize the enzymes of the respective compartments. Jaklitsch et al. (1991) reported that the CadA, which is e key enzyme for the biosynthesis of lA, is located in cytosol. Otiier important enzymes, such as citrate synthase and aconitase, are found in the mitochondria, but some residual level of these enzymes are also found in the cytosolic fraction. The depicted mechanism is that the ds-aconitate is transported to cytosol assisted by the malate-citrate antiporter. The biosynthetic pathway of LA in the citric acid cycle is illustrated in Fig. 10.5. 

When compounds enriched in the heavy-carbon isotope 13C and the radioactive carbon isotopes nC and 14C became available about 60 years ago, they were soon put to use in tracing the pathway of carbon atoms through the citric acid cycle. One such experiment initiated the controversy over the role of citrate. Acetate labeled in the carboxyl group (designated [1-14C] acetate) was incubated aerobically with an animal tissue preparation. Acetate is enzymatically converted to acetyl-CoA in animal tissues, and the pathway of the... 

Three modifications of the conventional oxidative citric acid cycle are needed, which substitute irreversible enzyme steps. Succinate dehydrogenase is replaced by fumarate reductase, 2-oxoglutarate dehydrogenase by ferredoxin-dependent 2-oxoglutarate oxidoreductase (2-oxoglutarate synthase), and citrate synthase by ATP-citrate lyase [3, 16] it should be noted that the carboxylases of the cycle catalyze the reductive carboxylation reactions. There are variants of the ATP-driven cleavage of citrate as well as of isocitrate formation [7]. The reductive citric acid... 

Pyruvate can be converted to acetyl-CoA via the pyruvate dehydrogenase complex. Pyruvate can also be carboxylated via pyruvate carboxylase to produce oxaloacetate. So, two molecules of pyruvate can form the precursors of citrate, which can be converted to succinate within the citric acid cycle. 

This finding ruled out citrate as a direct intermediate in the tricarboxylic acid cycle since isotopic citric acid, which is a symmetrical compound, could only give rise to a-ketoglutaric acid containing isotope distributed equally in both carboxyl groups (Fig. 5). Addition of citrate to their system (Evans and Slotin), furthermore, did not affect the specific radioactivity of the o-ketoglutarate. This effect would have been obtained had citrate been an intermediate. 

The second metabolic pathway which we have chosen to describe is the tricarboxylic acid cycle, often referred to as the Krebs cycle. This represents the biochemical hub of intermediary metabolism, not only in the oxidative catabolism of carbohydrates, lipids, and amino acids in aerobic eukaryotes and prokaryotes, but also as a source of numerous biosynthetic precursors. Pyruvate, formed in the cytosol by glycolysis, is transported into the matrix of the mitochondria where it is converted to acetyl CoA by the multi-enzyme complex, pyruvate dehydrogenase. Acetyl CoA is also produced by the mitochondrial S-oxidation of fatty acids and by the oxidative metabolism of a number of amino acids. The first reaction of the cycle (Figure 5.12) involves the condensation of acetyl Co and oxaloacetate to form citrate (1), a Claisen ester condensation. Citrate is then converted to the more easily oxidised secondary alcohol, isocitrate (2), by the iron-sulfur centre of the enzyme aconitase (described in Chapter 13). This reaction involves successive dehydration of citrate, producing enzyme-bound cis-aconitate, followed by rehydration, to give isocitrate. In this reaction, the enzyme distinguishes between the two external carboxyl groups... 

Aerobic glycolysis first involves a ten-step conversion of glucose to pyruvic acid or pyruvate, called the Embden-Meyerhoff-Pamas pathway, followed by its further conversion to carbon dioxide and water via what is variously called the tricarboxylic acid cycle, or citric acid cycle, or Krebs cycle after its discoverer. The net products discharged from the cycle are carbon dioxide and water, with recycle of a further product called oxaloacetic acid or oxaloacetate. Successive organic acids that contain three carboxyl groups (-COOH), are initially involved in the cycle starting with citric acid or a neutral salt of citric acid (citrate). Hence the designator tricarboxylic. 

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