Citric Acid Cycle

MUHAMMAD AWAIS

M PHIL BIOCHEMISTRY, PHARM-D

LECTURER, AIHS NAROWAL

Conversion of pyruvate to acetyl CoA
• Conversion of pyruvate to acetyl CoA occurs by oxidative decarboxylation.
• This is an irreversible reaction, catalysed by a multienzyme complex,
known as pyruvate dehydrogenase complex (PDH), which is found only
in the mitochondria.
• High activities of PDH are found in cardiac muscle and kidney.
• The enzyme PDH requires five cofactors (coenzymes), namely—TPP,
lipoamide, FAD, coenzyme A and NAD+
• The overall reaction of PDH is
PDH complex
• The PDH complex contains three enzymes
• Pyruvate dehydrogenase (E1)
• Dihydrolipoyl transacetylase (E2) and
• Dihydrolipoyl dehydrogenase (E3)
• Each present in multiple copies.
PDH complex reactions
• The sequence of reactions brought about by different enzymes of PDH
complex
• Pyruvate is decarboxylated to give hydroxyethyl TPP, catalysed by PDH
(decarboxylase activity)
• Dihydrolipoyl transacetylase brings about the formation of acetyl
lipoamide (from hydroxethyl-TPP) and then catalyses the transfer of acetyl
group to coenzyme A to produce acetyl CoA.
• The cycle is complete when reduced lipoamide is converted to oxidized
lipoamide by dihydrolipoyl dehydrogenase, transferring the reducing
equivalents to FAD.
• FADH2, in turn, transfers the reducing equivalents to NAD+ to give NADH +
H+, which can pass through the respiratory chain to give 3 ATP
PDH complex regulation
• Pyruvate dehydrogenase is a
good example for end product
(acetyl CoA, NADH) inhibition.
• PDH is also regulated by
phosphorylation and
dephosphorylation
• PDH is active as a
dephosphoenzyme while it is
inactive as a phosphoenzyme.
Citric acid cycle
• Krebs cycle or tricarboxylic acid—TCA cycle
• It is the most important metabolic pathway for the energy supply to the
body.
• About 65-70% of the ATP is synthesized in Krebs cycle.
• Citric acid cycle essentially involves the oxidation of acetyl CoA to
CO2 and H2O.
• This cycle utilizes about two thirds of total oxygen consumed by the body.
• There is no direct participation of oxygen in Krebs cycle. However, the
cycle operates only under aerobic conditions.
• The name TCA cycle is used, since, at the outset of the cycle, tricarboxylic
acids (citrate, cisaconitate and isocitrate) participate.
TCA cycle - central metabolic pathway
• The citric acid cycle is the final common oxidative pathway for
carbohydrates, fats and amino acids.
• This cycle not only supplies energy but also provides many
intermediates required for the synthesis of amino acids, glucose,
heme etc.
• Krebs cycle is the most important central pathway connecting
almost all the individual metabolic pathways (either directly or
indirectly).
TCA cycle
• The enzymes of TCA cycle are located in
mitochondrial matrix, in close
proximity to the electron transport chain.
This enables the synthesis of ATP by
oxidative phosphorylation without any
hindrance.
• Krebs cycle basically involves the
combination of a two carbon acetyl CoA
with a four carbon oxaloacetate to
produce a six carbon tricarboxylic
acid, citrate.
• The two carbons are oxidized to CO2
and oxaloacetate is regenerated and
Reactions of TCA cycle
1) Formation of citrate
• Krebs cycle proper starts with the condensation of acetyl CoA and
oxaloacetate, catalysed by the enzyme citrate synthase.
2a, 2b) Citrate is isomerized to isocitrate
• by the enzyme aconitase. This is achieved in a two stage reaction of
dehydration followed by hydration through the formation of an
intermediate—cis-aconitate.
Reactions of TCA cycle
3) Formation of alpha-ketoglutarate
• The enzyme isocitrate dehydrogenase (ICD) catalyses the conversion
(oxidative decarboxylation) of isocitrate to Alpha-ketoglutarate. The
formation of NADH and the liberation of CO2 occur at this stage.
4) Conversion of alpha-ketoglutarate to succinyl CoA
• occurs through oxidative decarboxylation, catalysed by alpha-
ketoglutarate dehydrogenase complex. This enzyme is dependent on five
cofactors—TPP, lipoamide, NAD+, FAD and CoA. The mechanism of the
reaction is analogous to the conversion of pyruvate to acetyl CoA
Reactions of TCA cycle
5) Formation of succinate
• Succinyl CoA is converted to succinate by succinate thiokinase. This
reaction is coupled with the phosphorylation of GDP to GTP. This is a
substrate level phosphorylation.
6) Conversion of succinate to fumarate :
• Succinate is oxidized by succinate dehydrogenase to fumarate. This
reaction results in the production of FADH2 and not NADH.
Reactions of TCA cycle
7) Formation of malate
• The enzyme fumarase catalyses the conversion of fumarate to malate with
the addition of H2O.
8) Conversion of malate to oxaloacetate
• Malate is then oxidized to oxaloacetate by malate dehydrogenase. The third
and final synthesis of NADH occurs at this stage. The oxaloacetate is
regenerated which can combine with another molecule of acetyl CoA,
and continue the cycle
• TCA cycle can be summarized as
Thank you 