Reductive TCA cycle

A reversed, reductive TCA cycle would require energy input to drive it. What might have been the thermodynamic driving force for such a cycle Wachtershanser hypothesizes that the anaerobic reaction of FeS and H9S to form insoluble FeS9 (pyrite, also known as fool s gold) in the prebiotic milieu could have been the driving reaction ... 

The only cultivated AOA, Nitrospumilus maritimus, depends on CO2 as its only carbon source and the presence of even low levels of organic carbon were inhibitory to growth. The pathway of CO2 fixation is, however, unknown. Hyperthermophilic Crenarchaeota generally utilize a 3-hydroxypropionate pathway or a reductive TCA cycle for autotrophic carbon fixation. Another cultivated marine Crenarchaeota strain, Cenarchaeum symbiosum, a sponge symbiont, appears to use the 3-hydroxypropionate pathway. It cannot be concluded on this basis which pathway is used by the AOA, but it very likely that is not the Calvin cycle. N. maritimus had a minimal generation time of 21 h, longer but roughly on the same scale as AOB. 

Grice et al., 1996b). This isotopic enrichment is typical for the reductive TCA cycle, the pathway of CO2 fixation followed by Chlorobiaceae (Section 8.03.6.1.4). 

Reductive TCA cycle in an aerobic bacterium, Hydrogenobacter thermophilus strain TK-6... 

Purification and characterization of components involved in the reductive TCA cycle in H. thermophilus strain TK-6 confirmed the operation of the cycle. 

Figure 18.1. The reductive TCA cycle (bold roman font) as an engine of synthesis of the major classes of biomolecules (bold italic). Synthesis of categories usually begins with a specific molecule (lightface).

The reductive TCA cycle uses CO2 for producing acetyl-CoA [28, 29]. Two CO2 units are converted in each cycle into acetyl-CoA using one ATP and four NAD(P) H units. The full cycle was first reported to be found in a green sulfur photosynthetic bacterium Chlorobium limicola) and was later also found to operate in Aquificales, Archeal Crenarcheota, and various types of proteobacteria. 

Figure 17.15 Major metabolic pathways involved in SA production in Saccbaromyces cerevisiae. Bold arrows indicate the major routes for succinate synthesis starting from glucose (a) via the reductive TCA cycle and (b) via the giyoxyiate cycle. PEP, phos-phoenolpyruvate OAA, oxaloacetate MAL, malate FUM, fumarate Suc-CoA, sucdnyl-CoA cr-KG, cr-ketoglutarate ICT, isodtrate CIT, citrate, ppc, PEP carboxykinase pyc, pyruvate carboxylase pyk, pyruvate kinase ...

The TCA cycle can now be completed by converting succinate to oxaloacetate. This latter process represents a net oxidation. The TCA cycle breaks it down into (consecutively) an oxidation step, a hydration reaction, and a second oxidation step. The oxidation steps are accompanied by the reduction of an [FAD] and an NAD. The reduced coenzymes, [FADHg] and NADH, subsequently provide reducing power in the electron transport chain. (We see in Chapter 24 that virtually the same chemical strategy is used in /3-oxidation of fatty acids.)... 

One of these alternate models, postulated by Gunter Wachtershanser, involves an archaic version of the TCA cycle running in the reverse (reductive) direction. Reversal of the TCA cycle results in assimilation of CO9 and fixation of carbon as shown. For each turn of the reversed cycle, two carbons are fixed in the formation of isocitrate and two more are fixed in the reductive transformation of acetyl-CoA to oxaloacetate. Thus, for every succinate that enters the reversed cycle, two succinates are returned, making the cycle highly antocatalytic. Because TCA cycle intermediates are involved in many biosynthetic pathways (see Section 20.13), a reversed TCA cycle would be a bountiful and broad source of metabolic substrates. 

Complex II is perhaps better known by its other name—succinate dehydrogenase, the only TCA cycle enzyme that is an integral membrane protein in the inner mitochondrial membrane. This enzyme has a mass of approximately 100 to 140 kD and is composed of four subunits two Fe-S proteins of masses 70 kD and 27 kD, and two other peptides of masses 15 kD and 13 kD. Also known as flavoprotein 2 (FP2), it contains an FAD covalently bound to a histidine residue (see Figure 20.15), and three Fe-S centers a 4Fe-4S cluster, a 3Fe-4S cluster, and a 2Fe-2S cluster. When succinate is converted to fumarate in the TCA cycle, concomitant reduction of bound FAD to FADHg occurs in succinate dehydrogenase. This FADHg transfers its electrons immediately to Fe-S centers, which pass them on to UQ. Electron flow from succinate to UQ,... 

Wachtcrshauser s prime candidate for a carbon-fixing process driven by pyrite formation is the reductive citrate cycle (RCC) mentioned above. Expressed simply, the RCC is the reversal of the normal Krebs cycle (tricarboxylic acid cycle TCA cycle), which is referred to as the turntable of metabolism because of its vital importance for metabolism in living cells. The Krebs cycle, in simplified form, can be summarized as follows ... 

A carrier molecule containing four carbon atoms (the C4 unit) takes up a C2 unit (the activated acetic acid ), which is introduced into the cycle. The product is a six-carbon molecule (the C6 unit), citric acid, or its salt, citrate. CO2 is cleaved off in a cyclic process, so that a C5 unit is left this loses a further molecule of CO2 to give the C4 unit, oxalacetate. In the living cell, this process involves ten steps, which are catalysed by eight enzymes. However, the purpose of the TCA cycle is not the elimination of CO2, but the provision of reduction equivalents, i.e., of electrons, and... 

If nitrogen (in the form of ammonia) is growth limiting, the potential applications of acetyl-CoA and NAD(P)H are restricted. Liberated NAD(P)H cannot be consumed for reductive syntheses, for instance of amino acids, it remains available and starts to inhibit citrate synthase [45, 46]. To the extent that the TCA cycle is thereby inhibited, acetyl-CoA should become available for the 3-ketothiolase, and could flow into poly(3HB) (Fig. 1, Table 1). 

The situation is simpler for odd numbered fatty acyl derivatives as [3-oxidation proceeds normally until a 5-carbon unit remains, rather than the usual 4-carbon unit. The C5 moiety is cleaved to yield acetyl-CoA (C2) and propionyl-CoA (C3). Propionyl CoA can be converted to succinyl CoA and enter the TCA cycle so the entire molecule is utilized but with a slight reduction in ATP yield as the opportunity to generate two molecules of NADH by isocitrate dehydrogenase and 2-oxoglutarate dehydrogenase is lost because succinyl-CoA occurs after these steps in the Krebs cycle (Figure 7.18). 

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