THE CITRIC ACID CYCLE TRICARBOXYLIC ACID(TCA) CYCLE KREBS CYCLE

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1 THE CITRIC ACID CYCLE TRICARBOXYLIC ACID(TCA) CYCLE KREBS CYCLE Production of Acetyl-CoA Reactions of the citric acid cycle Regulation of the citric acid cycle

2 Sir Hans Adolf Krebs A medical doctor and then professor of biochemistry Awarded Nobel Prize in Discovered the citric acid cycle also called TCA or Krebs cycle.

3 The Citric acid cycle)krebs cycle) The Citric acid cycle is universal central catabolic pathway in which compounds derived from the breakdown of carbohydrates, fats, proteins are oxidized to CO 2. Acetyl-CoA enters the citric acid cycle and condensate with oxaloacetate to citric acid. The citric acid cycle has 8 steps. 3 NADH + FADH 2 and GTP (ATP) are produced. Gives rise of 4C and 5C intermediates

4

5 Pyruvate is oxidized to acetyl-coa and CO 2 by pyruvate dehydrogenase complex (a cluster of enzymes, multiple copies of each of 3 enzymes located in mitochondria). Pyruvate dehydrogenase catalyzed an oxidation decarboxylation. Requires five coenzymes 1- TPP, 2- FAD, 3- Coenzyme A, 4- NAD, 5-Lipoate. Pyruvate dehydrogenase complex consists of enzymes 1- pyruvate dehydrogenase (E 1 ) 2- dihydrolipoyl transacetylase (E 2 ) 3- dihydrolipoyl dehydrogenase (E 3 )

6 Coenzyme A chemical structure the SH (thiol group of the mercaptoethylamine moiety form a thioester with acetate in acetyl CoA

7 Oxidation decarboxylation of pyruvate to acetyl-coa by pyruvate dehydrogenase E 1 Catalyzes first the decarboxylation 0f pyruvate producing hydroxyethyl-tpp, then the oxidation of the hydroxyethyl group to an acetyl group. E 2 Catalyzes the transfer of the acetyl group to coenzyme A forming acetyl-coa. E 3 Catalyzes the regeneration of disulfide (oxidized) form of lipoate electrons pass to FAD then to NAD.

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10 1) Formation of citrate The first step of cycle. Condensation of acetyl-coa with oxaloacetate to form citrate, catalyzed citrate synthase. Note 2C + 4C 6C with 3 carboxyl group COO -

11 2) Formation of isocitrate via cis-aconitate Dehydration and hydration step. Aconitase catalyzes reversible transformation of citrate to isocitrate. Cis-Aconitate is intermediate which does not dissociate from active site. Aconitase can promote the reversible addition of H 2 O to double bond in cis-aconitate lead to isocitrate.

12 3) Oxidation of isocitrate to α-ketoglutarate and CO 2 Oxidation decarboxylation step. Isocitrate dehydrogenase catalyzes irreversible oxidation decarboxylation of isocitrate to α-ketoglutarate. There are 2 form of Isocitrate dehydrogenase, one requiring NAD + as electron acceptor and other requiring NADP +. NADPH may be generated which essential for reductive anabolic reaction.

13 4) Oxidation of α ketoglutarate to succinyl-coa and CO 2 Oxidation decarboxylation step. α-ketoglutarate is converted to succinyl-coa and CO 2 by the action of α-ketoglutarate dehydrogenase complex. NAD + serves as electron acceptor. CoA serves as carrier of the succinyl group. α-ketoglutarate dehydrogenase resembles the pyruvate dehydrogenase in both structure and function.

14 5) Conversion of succinyl-coa to succinate Substrate level phosphorylation step. Reversible step. Succinyl-CoA synthetase (or succinic thiokinase) catalyzes conversion of succinyl-coa to succinate with production of GTP.

15 6) Oxidation of succinate to fumarate Dehydrogenation step. Reversible step. Succinate dehydrogenase catalyzed the oxidation of succinate to fumarate with forming of FADH 2. Succinate dehydrogenase is tightly bond to inner mitochondrial membrane.

16 7) Hydration of fumarate to malate Hydration step. Reversible step. Fumarase catalyzes the hydration of fumarate to malate. Fumarase is highly stereospecific,it catalyzes hydration of trans double bond of fumarate but not the cis double bond of maleate(the cis isomer of fumarate)

17 8) Oxidation of malate to oxaloacetate The last reaction of the citric acid cycle. Reversible step. Oxidation dehydration step. NAD-linked L-malate dehydrogenase catalyzes the oxidation of L-malate to oxaloacetate (4C) NADH produced.

18 Products of one turn of citric acid cycle 3 NADH 1 FADH 2 1 GTP 2 CO 2

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21 Regulation of metabolite flow from the pyruvate dehydrogenase complex through the citric cycle

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