The Krebs cycle (also known as the citric acid cycle or TCA cycle) is a key metabolic

pathway that plays a crucial role in cellular respiration. It occurs in the mitochondrial
matrix of eukaryotic cells and the cytoplasm of prokaryotes. The main function of the Krebs
cycle is to generate energy through the oxidation of acetyl-CoA, a derivative of
carbohydrates, fats, and proteins. This energy is stored in the form of high-energy molecules
such as NADH, FADH₂, and ATP.

Overview of the Krebs Cycle:

The Krebs cycle begins with the molecule acetyl-CoA entering the cycle and ultimately
results in the production of energy-rich molecules. These molecules are then used to produce
ATP, the primary energy currency of the cell. The cycle consists of a series of enzyme-
catalyzed reactions. Each turn of the cycle processes one molecule of acetyl-CoA, and since
two molecules of acetyl-CoA are produced per glucose molecule (through glycolysis), the
Krebs cycle runs twice for each glucose molecule.

Detailed Step-by-Step Process:

1. Formation of Citrate (Step 1):

o The cycle begins with acetyl-CoA (a 2-carbon molecule) combining with
oxaloacetate (a 4-carbon molecule).
o This forms citrate (a 6-carbon molecule), catalyzed by the enzyme citrate
synthase.
o Reaction: Acetyl-CoA + Oxaloacetate → Citrate
2. Formation of Isocitrate (Step 2):
o Citrate undergoes a reversible isomerization reaction. The enzyme aconitase
catalyzes the conversion of citrate into isocitrate, a rearranged form of citrate.
o Reaction: Citrate → Isocitrate
3. Oxidation to Alpha-Ketoglutarate (Step 3):
o Isocitrate is oxidized and decarboxylated (loses a CO₂ molecule) to form
alpha-ketoglutarate (a 5-carbon molecule).
o During this step, NAD⁺ is reduced to NADH.
o The enzyme responsible for this is isocitrate dehydrogenase.
o Reaction: Isocitrate → Alpha-Ketoglutarate + CO₂ + NADH
4. Formation of Succinyl-CoA (Step 4):
o Alpha-ketoglutarate undergoes another decarboxylation reaction, forming
succinyl-CoA (a 4-carbon molecule).
o This step also produces NADH from NAD⁺ and releases another molecule of
CO₂.
o The enzyme alpha-ketoglutarate dehydrogenase catalyzes this reaction.
o Reaction: Alpha-Ketoglutarate → Succinyl-CoA + CO₂ + NADH
5. Formation of Succinate (Step 5):
o Succinyl-CoA is converted into succinate. During this reaction, the high-
energy bond in succinyl-CoA is broken to form GTP (which is readily
converted to ATP).
o This is a substrate-level phosphorylation, where energy is directly used to
form ATP (or GTP, depending on the cell type).
o The enzyme involved is succinyl-CoA synthetase.
o Reaction: Succinyl-CoA → Succinate + GTP (or ATP)
 6. Oxidation to Fumarate (Step 6):
o Succinate is oxidized to form fumarate.
o In this step, the enzyme succinate dehydrogenase reduces FAD to FADH₂
(another high-energy electron carrier).
o Reaction: Succinate → Fumarate + FADH₂
7. Hydration to Malate (Step 7):
o Fumarate undergoes a hydration reaction, where water is added, forming
malate.
o The enzyme responsible is fumarase.
o Reaction: Fumarate → Malate
8. Oxidation to Oxaloacetate (Step 8):
o Finally, malate is oxidized to regenerate oxaloacetate (a 4-carbon molecule)
and reduce NAD⁺ to NADH.
o The enzyme malate dehydrogenase catalyzes this reaction.
o Reaction: Malate → Oxaloacetate + NADH

After the completion of these 8 steps, oxaloacetate is regenerated, and the cycle is ready to
begin again if another molecule of acetyl-CoA enters.

Products of the Krebs Cycle (per 1 turn):

 3 NADH (used in the electron transport chain for ATP production)

 1 FADH₂ (also used in the electron transport chain)
 1 GTP (or ATP) (direct energy transfer)
 2 CO₂ (waste products of the cycle)
 1 oxaloacetate (which is regenerated and can combine with another acetyl-CoA to
continue the cycle)

Significance of the Krebs Cycle:

 The Krebs cycle is the central hub of metabolism. It oxidizes fuel molecules (such as
glucose, fatty acids, and amino acids) to produce high-energy electron carriers, which
are later used in the electron transport chain to generate ATP, the primary energy
source for the cell.
 The cycle also provides important intermediates that can be used for biosynthesis
(e.g., amino acids, nucleotides, etc.).
 In addition to energy production, the Krebs cycle plays a role in maintaining the
balance of cellular metabolites and regulating various metabolic processes.

Summary of the Cycle:

The overall reaction of the Krebs cycle is:

Acetyl-CoA + 3 NAD⁺ + FAD + GDP + Pi + 2 H₂O → 2 CO₂ + 3 NADH + FADH₂ +

GTP + CoA

This simplified equation reflects the products generated per turn of the cycle.
By continuously cycling through these reactions, the Krebs cycle is integral in converting
energy stored in nutrients into usable ATP and maintaining cellular function.

The Krebs Cycle: Full Biochemical Breakdown

The Krebs cycle is a series of eight enzyme-catalyzed reactions that occur in the mitochondrial
matrix of eukaryotic cells (or in the cytoplasm of prokaryotes). It functions as the primary pathway
for the oxidation of acetyl-CoA (a derivative of carbohydrates, fats, and proteins) to generate high-
energy molecules that power cellular processes.

Step 1: Condensation of Acetyl-CoA and Oxaloacetate to Form Citrate

 Enzyme: Citrate Synthase

 Reaction:

Acetyl-CoA+Oxaloacetate→Citrate SynthaseCitrate+CoA\text{Acetyl-CoA} + \
text{Oxaloacetate} \xrightarrow{\text{Citrate Synthase}} \text{Citrate} + CoAAcetyl-
CoA+OxaloacetateCitrate SynthaseCitrate+CoA

o Acetyl-CoA, a 2-carbon molecule, combines with oxaloacetate, a 4-carbon molecule,

to form citrate, a 6-carbon molecule. This reaction is a condensation reaction
(forming a new carbon-carbon bond) and involves the hydrolysis of the thioester
bond in acetyl-CoA, releasing CoA as a byproduct.
o This is the first committed step of the cycle, where the acetyl group is incorporated
into the cycle.

Step 2: Isomerization of Citrate to Isocitrate

 Enzyme: Aconitase
 Reaction:

Citrate→AconitaseIsocitrate\text{Citrate} \xrightarrow{\text{Aconitase}} \
text{Isocitrate}CitrateAconitaseIsocitrate

o Citrate undergoes isomerization via a two-step process. First, citrate is dehydrated

to form cis-aconitate, and then water is added back to form isocitrate.
o This reaction is necessary to prepare citrate for the next oxidation step.
o Aconitase uses an iron-sulfur cluster to mediate the dehydration-hydration process.

Step 3: Oxidative Decarboxylation of Isocitrate to Alpha-Ketoglutarate

 Enzyme: Isocitrate Dehydrogenase

 Reaction:

Isocitrate→Isocitrate DehydrogenaseAlpha-Ketoglutarate+CO2+NADH\text{Isocitrate} \
xrightarrow{\text{Isocitrate Dehydrogenase}} \text{Alpha-Ketoglutarate} + CO_2 +
NADHIsocitrateIsocitrate DehydrogenaseAlpha-Ketoglutarate+CO2+NADH
 o Isocitrate is oxidized to alpha-ketoglutarate (a 5-carbon molecule) by isocitrate
dehydrogenase.
o In this reaction, isocitrate is first oxidized, transferring electrons to NAD⁺, forming
NADH.
o The resulting alpha-ketoglutarate is decarboxylated, releasing one molecule of CO₂
as a waste product.
o This is a rate-limiting step of the Krebs cycle.

Step 4: Oxidative Decarboxylation of Alpha-Ketoglutarate to Succinyl-CoA

 Enzyme: Alpha-Ketoglutarate Dehydrogenase Complex

 Reaction:

Alpha-Ketoglutarate+NAD++CoA→Alpha-Ketoglutarate Dehydrogenase ComplexSuccinyl-

CoA+CO2+NADH\text{Alpha-Ketoglutarate} + NAD^+ + CoA \xrightarrow{\text{Alpha-
Ketoglutarate Dehydrogenase Complex}} \text{Succinyl-CoA} + CO_2 + NADHAlpha-
Ketoglutarate+NAD++CoAAlpha-Ketoglutarate Dehydrogenase ComplexSuccinyl-CoA+CO2
+NADH

o Alpha-ketoglutarate, a 5-carbon molecule, is decarboxylated and oxidized by the

alpha-ketoglutarate dehydrogenase complex, a multienzyme complex similar to
pyruvate dehydrogenase.
o The enzyme complex involves NAD⁺ being reduced to NADH, and a CoA group being
attached to the remaining 4-carbon fragment, forming succinyl-CoA.
o A second CO₂ is released during this step.

Step 5: Substrate-Level Phosphorylation to Form Succinate

 Enzyme: Succinyl-CoA Synthetase

 Reaction:

Succinyl-CoA+GDP+Pi→Succinyl-CoA SynthetaseSuccinate+GTP+CoA\text{Succinyl-CoA} +
GDP + Pi \xrightarrow{\text{Succinyl-CoA Synthetase}} \text{Succinate} + GTP + CoASuccinyl-
CoA+GDP+PiSuccinyl-CoA SynthetaseSuccinate+GTP+CoA

o In this step, succinyl-CoA, a 4-carbon molecule, undergoes substrate-level

phosphorylation.
o The energy released from the breaking of the high-energy thioester bond in succinyl-
CoA is used to generate GTP (which can be converted to ATP).
o Succinate (a 4-carbon molecule) is produced, and the CoA group is released.
o This is the first step where ATP (or GTP, depending on the cell type) is directly
synthesized.

Step 6: Oxidation of Succinate to Fumarate

 Enzyme: Succinate Dehydrogenase

 Reaction:
 Succinate→Succinate DehydrogenaseFumarate+FADH2\text{Succinate} \xrightarrow{\
text{Succinate Dehydrogenase}} \text{Fumarate} +
FADH_2SuccinateSuccinate DehydrogenaseFumarate+FADH2

o Succinate (a 4-carbon molecule) is oxidized to fumarate (also a 4-carbon molecule)

by succinate dehydrogenase.
o In this reaction, FAD is reduced to FADH₂, which will later contribute electrons to the
electron transport chain for ATP production.
o Succinate dehydrogenase is unique in that it is part of both the Krebs cycle and the
electron transport chain (it is embedded in the inner mitochondrial membrane).

Step 7: Hydration of Fumarate to Malate

 Enzyme: Fumarase
 Reaction:

Fumarate+H2O→FumaraseMalate\text{Fumarate} + H_2O \xrightarrow{\text{Fumarase}} \

text{Malate}Fumarate+H2OFumaraseMalate

o Fumarate undergoes hydration, where water is added across the double bond,
forming malate.
o This is a simple addition of a water molecule, catalyzed by fumarase.

Step 8: Oxidation of Malate to Oxaloacetate

 Enzyme: Malate Dehydrogenase

 Reaction:

Malate→Malate DehydrogenaseOxaloacetate+NADH\text{Malate} \xrightarrow{\

text{Malate Dehydrogenase}} \text{Oxaloacetate} + NADHMalateMalate Dehydrogenase
Oxaloacetate+NADH

o Malate (a 4-carbon molecule) is oxidized to regenerate oxaloacetate (another 4-

carbon molecule), which is ready to combine with a new molecule of acetyl-CoA to
begin another round of the cycle.
o NAD⁺ is reduced to NADH in this final step of the cycle.

Products per Turn of the Krebs Cycle:

For each molecule of acetyl-CoA that enters the cycle, the following products are generated:

 3 NADH: These are used in the electron transport chain to produce ATP.
 1 FADH₂: This is also used in the electron transport chain for ATP production.
 1 GTP (or ATP): This is produced through substrate-level phosphorylation.
 2 CO₂: These are the waste products of the cycle and are expelled from the cell.
 1 Oxaloacetate: This is regenerated at the end of the cycle, allowing the process to continue.

The Overall Reaction of the Krebs Cycle (per acetyl-CoA):

Acetyl-CoA+3NAD++FAD+GDP+Pi+2H2O→2CO2+3NADH+FADH2+GTP+CoA\text{Acetyl-CoA} + 3
NAD^+ + FAD + GDP + Pi + 2 H_2O \rightarrow 2 CO_2 + 3 NADH + FADH_2 + GTP + CoAAcetyl-
CoA+3NAD++FAD+GDP+Pi+2H2O→2CO2+3NADH+FADH2+GTP+CoA

Since each glucose molecule generates 2 molecules of acetyl-CoA, the cycle runs twice for each
glucose molecule.

Importance of the Krebs Cycle:

1. Energy Production: The primary role of the Krebs cycle is to produce high-energy electron
carriers (NADH and FADH₂), which are used in the electron transport chain to generate ATP
through oxidative phosphorylation.
2. Intermediates for Biosynthesis: Many intermediates from the Krebs cycle serve as
precursors for the synthesis of amino acids, nucleotides, and other biomolecules.
3. Regulation of Metabolism: The cycle helps in the regulation of cellular energy levels. High
levels of ATP or NADH inhibit certain steps of the cycle, preventing excess production of
energy when it is not needed.

By continuously processing acetyl-CoA through the Krebs cycle, cells are able to efficiently harvest
energy from the oxidation of nutrients, ensuring proper cellular function and energy balance.