The citric acid cycle

Overview and steps of the citric acid cycle, also known as the Krebs cycle or
tricarboxylic acid (TCA) cycle.
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Introduction
How important is the citric acid cycle? So important that it
has not one, not two, but three different names in common
usage today!
The name we'll primarily use here, the citric acid cycle, refers
to the first molecule that forms during the cycle's reactions
citrate, or, in its protonated form, citric acid. However, you
may also hear this series of reactions called the tricarboxylic
acid (TCA) cycle, for the three carboxyl groups on its first two
intermediates, or the Krebs cycle, after its discoverer, Hans
Krebs.
[See a picture of the first two intermediates]

\text {H}^+H, start superscript, plus, end superscript

Whatever you prefer to call it, the citric cycle is a central

driver of cellular respiration. It takes
acetyl \text{CoA}CoAC, o, Aproduced by the oxidation of
pyruvate and originally derived from glucoseas its starting
material and, in a series of redox reactions, harvests much of

its bond energy in the form of \text{NADH}NADHN, A, D,

H,\text{FADH}_2FADH2F, A, D, H, start subscript, 2, end
subscript, and \text{ATP}ATPA, T, P molecules. The
reduced electron carriers\text{NADH}NADHN, A, D,
H and\text{FADH}_2FADH2F, A, D, H, start subscript, 2,
end subscriptgenerated in the TCA cycle will pass their
electrons into the electron transport chain and, through
oxidative phosphorylation, will generate most of the ATP
produced in cellular respiration.
Below, well look in more detail at how this remarkable cycle
works.

Overview of the citric acid cycle

In eukaryotes, the citric acid cycle takes place in the matrix
of the mitochondria, just like the conversion of pyruvate to
acetyl \text{CoA}CoAC, o, A. In prokaryotes, these steps
both take place in the cytoplasm. The citric acid cycle is a
closed loop; the last part of the pathway reforms the
molecule used in the first step. The cycle includes eight major
steps.

Simplified diagram of the citric acid cycle. First, acetyl CoA

combines with oxaloacetate, a four-carbon molecule, losing
the CoA group and forming the six-carbon molecule citrate.
After citrate undergoes a rearrangement step, it undergoes

an oxidation reaction, transferring electrons to NAD+ to form

NADH and releasing a molecule of carbon dioxide. The fivecarbon molecule left behind then undergoes a second, similar
reaction, transferring electrons to NAD+ to form NADH and
releasing a carbon dioxide molecule. The four-carbon
molecule remaining then undergoes a series of
transformations, in the course of which GDP and inorganic
phosphate are converted into GTPor, in some organisms,
ADP and inorganic phosphate are converted into ATPan FAD
molecule is reduced to FADH2, and another NAD+ is reduced
to NADH. At the end of this series of reactions, the fourcarbon starting molecule, oxaloacetate, is regenerated,
allowing the cycle to begin again.
In the first step of the cycle, acetyl

\text{CoA}CoAC, o,

A combines with a four-carbon acceptor molecule,

oxaloacetate, to form a six-carbon molecule called citrate.
After a quick rearrangement, this six-carbon molecule
releases two of its carbons as carbon dioxide molecules in a
pair of similar reactions, producing a molecule
of \text{NADH}NADHN, A, D, H each time^11start
superscript, 1, end superscript. The enzymes that catalyze
these reactions are key regulators of the citric acid cycle,
speeding it up or slowing it down based on the cells energy
needs^22start superscript, 2, end superscript.
The remaining four-carbon molecule undergoes a series of
additional reactions, first making an \text{ATP}ATPA, T,
P moleculeor, in some cells, a similar molecule

called \text{GTP}GTPG, T, Pthen reducing the electron

carrier \text{FAD}FADF, A, D to \text{FADH}_2FADH2F,
A, D, H, start subscript, 2, end subscript, and finally
generating another \text{NADH}NADHN, A, D, H. This set
of reactions regenerates the starting molecule, oxaloacetate,
so the cycle can repeat.
Overall, one turn of the citric acid cycle releases two carbon
dioxide molecules and produces
three \text{NADH}NADHN, A, D, H,
one \text{FADH}_2FADH2F, A, D, H, start subscript, 2, end
subscript, and one \text{ATP}ATPA, T,
P or \text{GTP}GTPG, T, P. The citric acid cycle goes
around twice for each molecule of glucose that enters cellular
respiration because there are two pyruvatesand thus, two
acetyl\text{CoA}CoAC, o, Asmade per glucose.

Steps of the citric acid cycle

You've already gotten a preview of the molecules produced
during the citric acid cycle. But how, exactly, are those
molecules made? Well walk through the cycle step by step,
seeing how \text{NADH}NADHN, A, D,
H, \text{FADH}_2FADH2F, A, D, H, start subscript, 2, end
subscript, and \text{ATP}ATPA, T, P/\text{GTP}GTPG, T,
P are produced and where carbon dioxide molecules are
released.
Step 1. In the first step of the citric acid cycle,
acetyl \text{CoA}CoAC, o, A joins with a four-carbon

molecule, oxaloacetate, releasing the

\text{CoA}CoAC, o,

A group and forming a six-carbon molecule called citrate.

Step 2. In the second step, citrate is converted into its
isomer, isocitrate. This is actually a two-step process,
involving first the removal and then the addition of a water
molecule, which is why the citric acid cycle is sometimes
described as having nine stepsrather than the eight listed
here^33start superscript, 3, end superscript.
Step 3. In the third step, isocitrate is oxidized and releases a
molecule of carbon dioxide, leaving behind a five-carbon
molecule-ketoglutarate. During this
step, \text{NAD}^+NAD+N, A, D, start superscript, plus,
end superscript is reduced to form \text{NADH}NADHN,
A, D, H. The enzyme catalyzing this step, isocitrate
dehydrogenase, is important in regulating the speed of the
citric acid cycle.
Step 4. The fourth step is similar to the third. In this case,
its -ketoglutarate thats oxidized,
reducing \text{NAD}^+NAD+N, A, D, start superscript,
plus, end superscript to \text{NADH}NADHN, A, D, H and
releasing a molecule of carbon dioxide in the process. The
remaining four-carbon molecule picks up Coenzyme A,
forming the unstable compound succinyl \text{CoA}CoAC,
o, A. The enzyme catalyzing this step, -ketoglutarate
dehydrogenase, is also important in regulation of the citric
acid cycle.

Detailed diagram of the citric acid cycle, showing the

structures of the various cycle intermediates and the
enzymes catalyzing each step.
Step 1. Acetyl CoA combines with oxaloacetate in a reaction
catalyzed by citrate synthase. This reaction also takes a water
molecule as a reactant, and it releases a SH-CoA molecule as
a product.
Step 2. Citrate is converted into isocitrate in a reaction
catalyzed by aconitase.
Step 3. Isocitrate is converted into -ketoglutarate in a
reaction catalyzed by isocitrate dehydrogenase. An NAD+
molecule is reduced to NADH + H+ in this reaction, and a
carbon dioxide molecule is released as a product.
Step 4. -ketoglutarate is converted to succinyl CoA in a
reaction catalyzed by -ketoglutarate dehydrogenase. An
NAD+ molecule is reduced to NADH + H+ in this reaction,
which also takes a SH-CoA molecule as reactant. A carbon
dioxide molecule is released as a product.
Step 5. Succinyl CoA is converted to succinate in a reaction
catalyzed by the enzyme succinyl-CoA synthetase. This
reaction converts inorganic phosphate, Pi, and GDP to GTP
and also releases a SH-CoA group.
Step 6. Succinate is converted to fumarate in a reaction
catalyzed by succinate dehydrogenase. FAD is reduced to
FADH2 in this reaction.

Step 7. Fumarate is converted to malate in a reaction

catalyzed by the enzyme fumarase. This reaction requires a
water molecule as a reactant.
Step 8. Malate is converted to oxaloacetate in a reaction
catalyzed by malate dehydrogenase. This reaction reduces an
NAD+ molecule to NADH + H+.
image credit: modified from "Oxidation of pyruvate and citric acid cycle: Figure 2" by
OpenStax College, Biology, CC BY 3.0

Step 5. In step five, the \text{CoA}CoAC, o, A of

succinyl \text{CoA}CoAC, o, A is replaced by a phosphate
group, which is then transferred to \text{ADP}ADPA, D,
P to make \text{ATP}ATPA, T, P. In some
cells,\text{GDP}GDPG, D, Pguanine diphosphateis
used instead of \text{ADP}ADPA, D, P,
forming \text{GTP}GTPG, T, Pguanine triphosphateas
a product. The four-carbon molecule produced in this step is
called succinate.
[Learn more about GTP.]

\text{GTP}G, T, P\text{ATP}A, T, P\text{ATP}A, T,

P\text{GTP}G, T, P
Step 6. In step six, succinate is oxidized, forming another
four-carbon molecule called fumarate. In this reaction, two
hydrogen atomswith their electronsare transferred
to \text{FAD}FADF, A, D,
producing \text{FADH}_2FADH2F, A, D, H, start subscript,
2, end subscript. The enzyme that carries out this step is
embedded in the inner membrane of the mitochondrion,
so \text{FADH}_2FADH2F, A, D, H, start subscript, 2, end

subscriptcan transfer its electrons directly into the electron

transport chain.
[Why use FAD here?]

\text{FAD}F, A, D\text{NAD}^+N, A, D, start

superscript, plus, end superscript\text{NAD}^+N, A, D,
start superscript, plus, end superscript\text{FAD}F, A,
D^{4,5}start superscript, 4, comma, 5, end superscript
Step 7. In step seven, water is added to the four-carbon
molecule fumarate, converting it into another four-carbon
molecule called malate.
Step 8. In the last step of the citric acid cycle, oxaloacetate
the starting four-carbon compoundis regenerated by
oxidation of malate. Another molecule
of \text{NAD}^+NAD+N, A, D, start superscript, plus, end
superscript is reduced to \text{NADH}NADHN, A, D, H in
the process.

Products of the citric acid cycle

Lets take a step back and do some accounting, tracing the
fate of the carbons that enter the citric acid cycle and
counting the reduced electron carriers
\text{NADH}NADHN, A, D,
H and \text{FADH}_2FADH2F, A, D, H, start subscript, 2,
end subscriptand \text{ATP}ATPA, T, P produced.
In a single turn of the cycle,

two carbons enter from acetyl

\text{CoA}CoAC, o, A,

and two molecules of carbon dioxide are released;

\text{NADH}NADHN, A, D, H and
one molecule of \text{FADH}_2FADH2F, A, D, H, start
three molecules of

subscript, 2, end subscript are generated; and

\text{ATP}ATPA, T,
P or \text{GTP}GTPG, T, P is produced.
one molecule of

These figures are for one turn of the cycle, corresponding to

one molecule of acetyl \text{CoA}CoAC, o, A. Each
glucose produces two acetyl \text{CoA}CoAC, o,
A molecules, so we need to multiply these numbers by

222 if

we want the per-glucose yield.

Two carbonsfrom acetyl

\text{CoA}CoAC, o, Aenter the

citric acid cycle in each turn, and two carbon dioxide

molecules are released. However, the carbon dioxide
molecules dont actually contain carbon atoms from the
acetyl \text{CoA}CoAC, o, A that just entered the cycle.
Instead, the carbons from acetyl \text{CoA}CoAC, o, A are
initially incorporated into the intermediates of the cycle and
are released as carbon dioxide only during later turns. After
enough turns, all the carbon atoms from the acetyl group of
acetyl \text{CoA}CoAC, o, A will be released as carbon
dioxide.

Wheres all the

P?

\text{ATP}ATPA, T,

You may be thinking that the

\text{ATP}ATPA, T, P output

of the citric acid cycle seems pretty unimpressive. All that

work for just one \text{ATP}ATPA, T,

P or \text{GTP}GTPG, T, P?
Its true that the citric acid cycle doesnt produce
much \text{ATP}ATPA, T, P directly. However, it can make
a lot of \text{ATP}ATPA, T, P indirectly, by way of
the \text{NADH}NADHN, A, D,
H and\text{FADH}_2FADH2F, A, D, H, start subscript, 2,
end subscript it generates. These electron carriers will
connect with the last portion of cellular respiration,
depositing their electrons into the electron transport chain to
drive synthesis of ATP molecules through oxidative
phosphorylation.

Citric acid cycle

From Wikipedia, the free encyclopedia

Overview of the citric acid cycle (click to enlarge)

The citric acid cycle also known as the tricarboxylic acid (TCA) cycle or the Krebs cycle[1][2] is
a series ofchemical reactions used by all aerobic organisms to generate energy through
the oxidation of acetyl-CoA derived from carbohydrates, fats and proteins into carbon dioxide and
chemical energy in the form of adenosine triphosphate. In addition, the cycle provides precursors of
certain amino acids as well as the reducing agent NADH that is used in numerous other biochemical
reactions. Its central importance to many biochemical pathways suggests that it was one of the
earliest established components of cellular metabolism and may have originated abiogenically.[3][4]
The name of this metabolic pathway is derived from citric acid (a type of tricarboxylic acid) that is
consumed and then regenerated by this sequence of reactions to complete the cycle. In addition, the
cycle consumes acetate (in the form of acetyl-CoA) and water, reduces NAD+ to NADH, and
produces carbon dioxide as a waste byproduct. The NADH generated by the TCA cycle is fed into
the oxidative phosphorylation (electron transport) pathway. The net result of these two closely linked
pathways is the oxidation of nutrients to produce usable chemical energy in the form of ATP.
In eukaryotic cells, the citric acid cycle occurs in the matrix of the mitochondrion. In prokaryotic cells,
such as bacteria which lack mitochondria, the TCA reaction sequence is performed in
the cytosol with the proton gradient for ATP production being across the cell's surface (plasma
membrane) rather than the inner membrane of the mitochondrion.
Contents
[hide]

1Discovery

2Evolution

3Overview

4Steps

5Products

6Efficiency

7Variation

8Regulation

9Major metabolic pathways converging on the TCA cycle

10Citric acid cycle intermediates serve as substrates for biosynthetic processes

11Interactive pathway map

12See also

13References

14External links

Discovery[edit]
Several of the components and reactions of the citric acid cycle were established in the 1930s by the
research of the Nobel laureate Albert Szent-Gyrgyi, for which he received the Nobel Prize in 1937
for his discoveries pertaining to fumaric acid, a key component of the cycle.[5] The citric acid cycle
itself was finally identified in 1937 by Hans Adolf Krebswhile at the University of Sheffield, for which
he received the Nobel Prize for Physiology or Medicine in 1953.[6]

Evolution[edit]
Components of the TCA cycle were derived from anaerobic bacteria, and the TCA cycle itself may
have evolved more than once.[7] Theoretically there are several alternatives to the TCA cycle;
however, the TCA cycle appears to be the most efficient. If several TCA alternatives had evolved
independently, they all appear to have converged to the TCA cycle.[8][9]

Overview[edit]

Structural diagram of acetyl-CoA. The portion in blue, on the left, is the acetyl group; the portion in black
is coenzyme A.

The citric acid cycle is a key metabolic pathway that unifies carbohydrate, fat, and protein
metabolism. The reactions of the cycle are carried out by 8 enzymes that completely oxidize acetate,
in the form of acetyl-CoA, into two molecules each of carbon dioxide and water.
Through catabolism of sugars, fats, and proteins, a two-carbon organic product acetate in the form
of acetyl-CoA is produced which enters the citric acid cycle. The reactions of the cycle also convert
three equivalents ofnicotinamide adenine dinucleotide (NAD+) into three equivalents of reduced

NAD+ (NADH), one equivalent of flavin adenine dinucleotide (FAD) into one equivalent of FADH2, and
one equivalent each of guanosine diphosphate (GDP) and inorganicphosphate (Pi) into one
equivalent of guanosine triphosphate (GTP). The NADH and FADH2 generated by the citric acid
cycle are in turn used by the oxidative phosphorylation pathway to generate energy-rich adenosine
triphosphate (ATP).
One of the primary sources of acetyl-CoA is from the breakdown of sugars by glycolysis which
yield pyruvate that in turn is decarboxylated by the enzyme pyruvate dehydrogenase generating
acetyl-CoA according to the following reaction scheme:

CH3C(=O)C(=O)O (pyruvate) + HSCoA + NAD+ CH3C(=O)SCoA (acetyl-CoA) + NADH +

CO2

The product of this reaction, acetyl-CoA, is the starting point for the citric acid cycle. Acetyl-CoA may
also be obtained from the oxidation of fatty acids. Below is a schematic outline of the cycle:

The citric acid cycle begins with the transfer of a two-carbon acetyl group from acetyl-CoA to
the four-carbon acceptor compound (oxaloacetate) to form a six-carbon compound (citrate).

The citrate then goes through a series of chemical transformations, losing

two carboxyl groups as CO2. The carbons lost as CO2 originate from what was oxaloacetate, not
directly from acetyl-CoA. The carbons donated by acetyl-CoA become part of the oxaloacetate
carbon backbone after the first turn of the citric acid cycle. Loss of the acetyl-CoA-donated
carbons as CO2 requires several turns of the citric acid cycle. However, because of the role of
the citric acid cycle in anabolism, they might not be lost, since many TCA cycle intermediates are
also used as precursors for the biosynthesis of other molecules.[10]

Most of the energy made available by the oxidative steps of the cycle is transferred as
energy-rich electrons to NAD+, forming NADH. For each acetyl group that enters the citric acid
cycle, three molecules of NADH are produced.

Electrons are also transferred to the electron acceptor Q, forming QH 2.

At the end of each cycle, the four-carbon oxaloacetate has been regenerated, and the cycle
continues.

Steps[edit]
Two carbon atoms are oxidized to CO2, the energy from these reactions being transferred to other
metabolic processes by GTP (or ATP), and as electrons in NADH and QH2. The NADH generated in
the TCA cycle may later donate its electrons oxidative phosphorylation to drive ATP
synthesis; FADH2 is covalently attached to succinate dehydrogenase, an enzyme functioning both in
the TCA cycle and the mitochondrial electron transport chain in oxidative phosphorylation. FADH2,
therefore, facilitates transfer of electrons tocoenzyme Q, which is the final electron acceptor of the
reaction catalyzed by the Succinate:ubiquinone oxidoreductase complex, also acting as an
intermediate in the electron transport chain.[11]
The citric acid cycle is continuously supplied with new carbon in the form of acetyl-CoA, entering at
step 0 below.[12]
Substrates

Products

Enzyme

Reaction

Comment

type

0
/
1
0

Oxaloacetat
e+
Acetyl
CoA +
H2O

Citrate +
CoA-SH

Citrate

cis-Aconitate
+
H2 O

Citrate
synthase

Aldol
condensation

irreversible,
extends the 4C oxaloacetate to a 6C
molecule

Dehydration
Aconitase

reversible isomerisation

cisAconitate +
H2O

Isocitrate

Hydration

Isocitrate +
NAD+

Oxalosuccina
te+
NADH + H +

Oxidation

generates NADH (equivalent of 2.5 ATP)

Decarboxylat
ion

rate-limiting, irreversible stage,

generates a 5C molecule

Oxalosuccin
ate

Ketoglutarate
+
CO2

Ketoglutarat
e+
NAD+ +
CoA-SH

SuccinylCoA +
NADH +
H+ +
CO2

Isocitrate
dehydrogen
ase

Ketoglutarat Oxidative
e
decarboxylati
dehydrogen on
ase

SuccinylCoA +
GDP + Pi

Succinate +
CoA-SH +
GTP

SuccinylCoA
synthetase

substratelevel
phosphorylati
on

Succinate +

Fumarate +

Succinate

Oxidation

irreversible stage,
generates NADH (equivalent of 2.5
ATP),
regenerates the 4C chain (CoA excluded)

or ADPATP instead of GDPGTP,[11]

generates 1 ATP or equivalent
Condensation
reaction of GDP + Pi and hydrolysis of S
uccinyl-CoA involve the H2O needed for
balanced equation.
uses FAD as a prosthetic

group (FADFADH2 in the first step of

the reaction) in the enzyme,[11]
generates the equivalent of 1.5 ATP

ubiquinone (
Q)

ubiquinol (Q
H2 )

dehydrogen
ase

Fumarate +
H2O

L-Malate

Fumarase

Hydration

Hydration of C-C double bond

L-Malate +
NAD+

Oxaloacetate
+
NADH + H+

Malate
dehydrogen
ase

Oxidation

reversible (in fact, equilibrium favors

malate), generates NADH (equivalent of
2.5 ATP)

1
0
/
0

Oxaloacetat
e+
Acetyl
CoA +
H2O

Citrate +
CoA-SH

Citrate
synthase

Aldol
condensation

This is the same as step 0 and restarts the

cycle. The reaction is irreversible and
extends the 4C oxaloacetate to a 6C
molecule

Mitochondria in animals, including humans, possess two succinyl-CoA synthetases: one that
produces GTP from GDP, and another that produces ATP from ADP.[13] Plants have the type that
produces ATP (ADP-forming succinyl-CoA synthetase).[12] Several of the enzymes in the cycle may
be loosely associated in a multienzyme protein complex within the mitochondrial matrix.[14]
The GTP that is formed by GDP-forming succinyl-CoA synthetase may be utilized by nucleosidediphosphate kinase to form ATP (the catalyzed reaction is GTP + ADP GDP + ATP).[11]

Products[edit]
Products of the first turn of the cycle are: one GTP (or ATP), three NADH, one QH2, two CO2.
Because two acetyl-CoA molecules are produced from each glucose molecule, two cycles are
required per glucose molecule. Therefore, at the end of two cycles, the products are: two GTP, six
NADH, two QH2, and four CO2
Description

Reactants

Products

The sum of all reactions in the citric acid cycle is:

Acetyl-CoA + 3
NAD+ + Q + GDP +
Pi + 2 H2O

CoA-SH + 3
NADH + 3 H+ +
QH2 + GTP + 2 CO2

Combining the reactions occurring during the pyruvate

oxidation with those occurring during the citric acid cycle,
the following overall pyruvate oxidation reaction is
obtained:

Pyruvate ion + 4
NAD+ + Q + GDP +
Pi + 2 H2O

4 NADH + 4 H+ +
QH2 + GTP + 3 CO2

Combining the above reaction with the ones occurring in

the course of glycolysis, the following overall glucose
oxidation reaction (excluding reactions in the respiratory
chain) is obtained:

Glucose + 10 NAD+ +
2 Q + 2 ADP + 2 GDP
+ 4 Pi + 2 H2O

10 NADH + 10
H+ + 2 QH2 + 2 ATP
+ 2 GTP + 6 CO2

The above reactions are balanced if Pi represents the H2PO4 ion, ADP and GDP the ADP2 and
GDP2 ions, respectively, and ATP and GTP the ATP3 and GTP3 ions, respectively.
The total number of ATP obtained after complete oxidation of one glucose in glycolysis, citric acid
cycle, and oxidative phosphorylation is estimated to be between 30 and 38.[15]

Efficiency[edit]
The theoretical maximum yield of ATP through oxidation of one molecule of glucose in glycolysis,
citric acid cycle, and oxidative phosphorylation is 38 (assuming 3 molar equivalents of ATP per
equivalent NADH and 2 ATP per FADH2). In eukaryotes, two equivalents of NADH are generated
in glycolysis, which takes place in the cytoplasm. Transport of these two equivalents into the
mitochondria consumes two equivalents of ATP, thus reducing the net production of ATP to 36.
Furthermore, inefficiencies in oxidative phosphorylation due to leakage of protons across the
mitochondrial membrane and slippage of the ATP synthase/proton pump commonly reduces the ATP
yield from NADH and FADH2 to less than the theoretical maximum yield.[15] The observed yields are,
therefore, closer to ~2.5 ATP per NADH and ~1.5 ATP per FADH2, further reducing the total net
production of ATP to approximately 30.[16] An assessment of the total ATP yield with newly revised
proton-to-ATP ratios provides an estimate of 29.85 ATP per glucose molecule. [17]

Variation[edit]
While the TCA cycle is in general highly conserved, there is significant variability in the enzymes
found in different taxa[18] (note that the diagrams on this page are specific to the mammalian pathway
variant).
Some differences exist between eukaryotes and prokaryotes. The conversion of D-threo-isocitrate to
2-oxoglutarate is catalyzed in eukaryotes by the NAD+-dependent EC 1.1.1.41, while prokaryotes
employ the NADP+-dependent EC 1.1.1.42.[19] Similarly, the conversion of (S)-malate to oxaloacetate
is catalyzed in eukaryotes by the NAD+-dependent EC 1.1.1.37, while most prokaryotes utilize a
quinone-dependent enzyme, EC 1.1.5.4.[20]
A step with significant variability is the conversion of succinyl-CoA to succinate. Most organisms
utilize EC 6.2.1.5, succinateCoA ligase (ADP-forming) (despite its name, the enzyme operates in
the pathway in the direction of ATP formation). In mammals a GTP-forming enzyme, succinateCoA
ligase (GDP-forming) (EC 6.2.1.4) also operates. The level of utilization of each isoform is tissue
dependent.[21] In some acetate-producing bacteria, such as Acetobacter aceti, an entirely different
enzyme catalyzes this conversion EC 2.8.3.18, succinyl-CoA:acetate CoA-transferase. This
specialized enzyme links the TCA cycle with acetate metabolism in these organisms. [22] Some
bacteria, such asHelicobacter pylori, employ yet another enzyme for this conversion succinylCoA:acetoacetate CoA-transferase (EC 2.8.3.5).[23]
Some variability also exists at the previous step the conversion of 2-oxoglutarate to succinyl-CoA.
While most organisms utilize the ubiquitous NAD+-dependent 2-oxoglutarate dehydrogenase, some
bacteria utilize a ferredoxin-dependent 2-oxoglutarate synthase (EC 1.2.7.3).[24] Other organisms,
including obligately autotrophic and methanotrophic bacteria and archaea, bypass succinyl-CoA
entirely, and convert 2-oxoglutarate to succinate via succinate semialdehyde, using EC 4.1.1.71, 2oxoglutarate decarboxylase, andEC 1.2.1.79, succinate-semialdehyde dehydrogenase.[25]

Regulation[edit]
The regulation of the TCA cycle is largely determined by product inhibition and substrate availability.
If the cycle were permitted to run unchecked, large amounts of metabolic energy could be wasted in
overproduction of reduced coenzyme such as NADH and ATP. The major eventual substrate of the
cycle is ADP which gets converted to ATP. A reduced amount of ADP causes accumulation of

precursor NADH which in turn can inhibit a number of enzymes. NADH, a product of all
dehydrogenases in the TCA cycle with the exception of succinate dehydrogenase, inhibits pyruvate
dehydrogenase, isocitrate dehydrogenase, -ketoglutarate dehydrogenase, and also citrate
synthase. Acetyl-coAinhibits pyruvate dehydrogenase, while succinyl-CoA inhibits alphaketoglutarate dehydrogenase and citrate synthase. When tested in vitro with TCA enzymes, ATP
inhibits citrate synthase and -ketoglutarate dehydrogenase; however, ATP levels do not change
more than 10% in vivo between rest and vigorous exercise. There is no known allostericmechanism
that can account for large changes in reaction rate from an allosteric effector whose concentration
changes less than 10%.[26]
Calcium is used as a regulator. Mitochondrial matrix calcium levels can reach the tens of micromolar
levels during cellular activation.[27] It activates pyruvate dehydrogenase phosphatase which in turn
activates the pyruvate dehydrogenase complex. Calcium also activates isocitrate
dehydrogenase and -ketoglutarate dehydrogenase.[28] This increases the reaction rate of many of
the steps in the cycle, and therefore increases flux throughout the pathway.
Citrate is used for feedback inhibition, as it inhibits phosphofructokinase, an enzyme involved
in glycolysis that catalyses formation of fructose 1,6-bisphosphate,a precursor of pyruvate. This
prevents a constant high rate of flux when there is an accumulation of citrate and a decrease in
substrate for the enzyme.
Recent work has demonstrated an important link between intermediates of the citric acid cycle and
the regulation of hypoxia-inducible factors (HIF). HIF plays a role in the regulation of oxygen
homeostasis, and is a transcription factor that targets angiogenesis, vascular remodeling, glucose
utilization, iron transport and apoptosis. HIF is synthesized consititutively, and hydroxylation of at
least one of two critical proline residues mediates their interaction with the von Hippel Lindau E3
ubiquitin ligase complex, which targets them for rapid degradation. This reaction is catalysed
by prolyl 4-hydroxylases. Fumarate and succinate have been identified as potent inhibitors of prolyl
hydroxylases, thus leading to the stabilisation of HIF.[29]

Major metabolic pathways converging on the TCA cycle[edit]

Several catabolic pathways converge on the TCA cycle. Most of these reactions add intermediates to
the TCA cycle, and are therefore known as anaplerotic reactions, from the Greek meaning to "fill up".
These increase the amount of acetyl CoA that the cycle is able to carry, increasing the
mitochondrion's capability to carry out respiration if this is otherwise a limiting factor. Processes that
remove intermediates from the cycle are termed "cataplerotic" reactions.
In this section and in the next, the citric acid cycle intermediates are indicated in italics to distinguish
them from other substrates and end-products.
Pyruvate molecules produced by glycolysis are actively transported across the
inner mitochondrial membrane, and into the matrix. Here they can be oxidized and combined
withcoenzyme A to form CO2, acetyl-CoA, and NADH, as in the normal cycle.[30]
However, it is also possible for pyruvate to be carboxylated by pyruvate carboxylase to
form oxaloacetate. This latter reaction "fills up" the amount of oxaloacetate in the citric acid cycle,
and is therefore an anaplerotic reaction, increasing the cycles capacity to metabolize acetylCoA when the tissue's energy needs (e.g. in muscle) are suddenly increased by activity.[31]
In the citric acid cycle all the intermediates (e.g. citrate, iso-citrate, alpha-ketoglutarate, succinate,
fumarate, malate and oxaloacetate) are regenerated during each turn of the cycle. Adding more of
any of these intermediates to the mitochondrion therefore means that that additional amount is
retained within the cycle, increasing all the other intermediates as one is converted into the other.
Hence the addition of any one of them to the cycle has an anaplerotic effect, and its removal has a
cataplerotic effect. These anaplerotic and cataplerotic reactions will, during the course of the cycle,
increase or decrease the amount of oxaloacetate available to combine with acetyl-CoA to form citric

acid. This in turn increases or decreases the rate of ATP production by the mitochondrion, and thus
the availability of ATP to the cell.[31]
Acetyl-CoA, on the other hand, derived from pyruvate oxidation, or from the beta-oxidation of fatty
acids, is the only fuel to enter the citric acid cycle. With each turn of the cycle one molecule
of acetyl-CoA is consumed for every molecule of oxaloacetate present in the mitochondrial matrix,
and is never regenerated. It is the oxidation of the acetate portion of acetyl-CoA that produces
CO2 and water, with the energy thus released captured in the form of ATP.[31]
In the liver, the carboxylation of cytosolic pyruvate into intra-mitochondrial oxaloacetate is an early
step in the gluconeogenic pathway which converts lactate and de-aminatedalanine into glucose,[30]
[31]
under the influence of high levels of glucagon and/or epinephrine in the blood.[31] Here the addition
of oxaloacetate to the mitochondrion does not have a net anaplerotic effect, as another citric acid
cycle intermediate (malate) is immediately removed from the mitochondrion to be converted into
cytosolic oxaloacetate, which is ultimately converted into glucose, in a process that is almost the
reverse of glycolysis.[31]
In protein catabolism, proteins are broken down by proteases into their constituent amino acids.
Their carbon skeletons (i.e. the de-aminated amino acids) may either enter the citric acid cycle as
intermediates (e.g. alpha-ketoglutarate derived from glutamate or glutamine), having an anaplerotic
effect on the cycle, or, in the case of leucine, isoleucine, lysine, phenylalanine, tryptophan, and
tyrosine, they are converted into acetyl-CoA which can be burned to CO2 and water, or used to
form ketone bodies, which too can only be burned in tissues other than the liver where they are
formed, or excreted via the urine or breath.[31] These latter amino acids are therefore termed
"ketogenic" amino acids, whereas those that enter the citric acid cycle as intermediates can only be
cataplerotically removed by entering the gluconeogenic pathway via malate which is transported out
of the mitochondrion to be converted into cytosolic oxaloacetate and ultimately into glucose. These
are the so-called "glucogenic" amino acids. De-aminated alanine, cysteine, glycine, serine, and
threonine are converted to pyruvate and can consequently either enter the citric acid cycle
as oxaloacetate (an anaplerotic reaction) or as acetyl-CoA to be disposed of as CO2 and water.[31]
In fat catabolism, triglycerides are hydrolyzed to break them into fatty acids and glycerol. In the liver
the glycerol can be converted into glucose via dihydroxyacetone phosphateand glyceraldehyde-3phosphate by way of gluconeogenesis. In many tissues, especially heart and skeletal muscle tissue,
fatty acids are broken down through a process known as beta oxidation, which results in the
production of mitochondrial acetyl-CoA, which can be used in the citric acid cycle. Beta oxidation of
fatty acids with an odd number ofmethylene bridges produces propionyl-CoA, which is then
converted into succinyl-CoA and fed into the citric acid cycle as an anaplerotic intermediate.[32]
The total energy gained from the complete breakdown of one (six-carbon) molecule of glucose
by glycolysis, the formation of 2 acetyl-CoA molecules, their catabolism in the citric acid cycle, and
oxidative phosphorylation equals about 30 ATP molecules, in eukaryotes. The number of ATP
molecules derived from the beta oxidation of a 6 carbon segment of a fatty acid chain, and the
subsequent oxidation of the resulting 3 molecules of acetyl-CoA is 40.

Citric acid cycle intermediates serve as substrates for biosynthetic

processes[edit]
In this subheading, as in the previous one, the TCA intermediates are identified by italics.
Several of the citric acid cycle intermediates are used for the synthesis of important compounds,
which will have significant cataplerotic effects on the cycle. [31] Acetyl-CoA cannot be transported out
of the mitochondrion. To obtain cytosolic acetyl-CoA, citrate is removed from the citric acid cycle and
carried across the inner mitochondrial membrane into the cytosol. There it is cleaved by ATP citrate
lyase into acetyl-CoA and oxaloacetate. The oxaloacetate is returned to mitochondrion
as malate (and then converted back intooxaloacetate to transfer more acetyl-CoA out of the

mitochondrion).[33] The cytosolic acetyl-CoA is used for fatty acid synthesis and the production of
cholesterol. Cholesterol can, in turn, be used to synthesize the steroid hormones, bile salts,
and vitamin D.[30][31]
The carbon skeletons of many non-essential amino acids are made from citric acid cycle
intermediates. To turn them into amino acids the alpha keto-acids formed from the citric acid cycle
intermediates have to acquire their amino groups from glutamate in a transamination reaction, in
which pyridoxal phosphate is a cofactor. In this reaction the glutamate is converted into alphaketoglutarate, which is a citric acid cycle intermediate. The intermediates that can provide the carbon
skeletons for amino acid synthesis are oxaloacetatewhich forms aspartate and asparagine;
and alpha-ketoglutarate which forms glutamine, proline, and arginine.[30][31]
Of these amino acids, aspartate and glutamine are used, together with carbon and nitrogen atoms
from other sources, to form the purines that are used as the bases in DNA andRNA, as well as
in ATP, AMP, GTP, NAD, FAD and CoA.[31]
The pyrimidines are partly assembled from aspartate (derived from oxaloacetate). The
pyrimidines, thymine, cytosine and uracil, form the complementary bases to the purine bases in DNA
and RNA, and are also components of CTP, UMP, UDP and UTP.[31]
The majority of the carbon atoms in the porphyrins come from the citric acid cycle
intermediate, succinyl-CoA. These molecules are an important component of the hemoproteins,
such as hemoglobin, myoglobin and various cytochromes.[31]
During gluconeogenesis mitochondrial oxaloacetate is reduced to malate which is then transported
out of the mitochondrion, to be oxidized back to oxaloacetate in the cytosol. Cytosolic oxaloacetate
is then decarboxylated to phosphoenolpyruvate by phosphoenolpyruvate carboxykinase, which is
the rate limiting step in the conversion of nearly all the gluconeogenic precursors (such as the
glucogenic amino acids and lactate) into glucose by the liver and kidney.[30][31]
Because the citric acid cycle is involved in both catabolic and anabolic processes, it is known as
an amphibole pathway.

Interactive pathway map[edit]

Click on genes, proteins and metabolites below to link to respective articles.
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File:TCACycle WP78.png

TCA Cycle edit

1.

Jump up^ The interactive pathway map can be edited at WikiPathways: "TCACycle_WP78".