TRICARBOXYLIC ACID CYCLE

Tricarboxylic acid cycle, (TCA cycle), also called Krebs cycle and citric acid cycle, the second
stage of cellular respiration, the three-stage process by which living cells break down organic fuel
molecules in the presence of oxygen to harvest the energy they need to grow and divide The
pathway is sometimes known as the citric acid cycle, or the Krebs' cycle, after its discoverer, Sir
Hans Krebs. In addition to its role in energy-yielding metabolism, and the oxidation of 2-carbon
units, the cycle is also the major pathway for inter conversion of 4- and 5-carbon compounds in
the cell, many of which arise from, or are intermediates in the synthesis of, amino acids.
Oxaloacetate, a key intermediate in the cycle, is the main precursor for gluconeogenesis in the
fasting state.

Tricarboxylic Acid Cycle

The Discoverer of the Cycle
The tricarboxylic acid (TCA) cycle was first correctly formulated by Han Krebs, who was born on
25 August 1900 in Hildesheim. He was the son of a surgeon, Georg, and mother Alma, who was to
die of depression when Krebs was 19. After completing gymnasium in Hildesheim in September
1918, Krebs joined the German army signal corps until the armistice of 11 November

1918. After Completing his training in internal medicine at Gottingen, Freiburg, Munich, and
Berlin in December 1925, Krebs looked for ways to obtain biochemical training. The time spent
in the clinics had convinced him that it was necessary to pursue medical research. Through the
influence of Bruno Mendel, who served with Krebs in the Third Medical Clinic in Berlin, and who
was an acquaintance of Albert Einstein, professor of Physics at the University of Berlin, who was
in turn a frequent dinner guest at the home of Otto Warburg's father, Mendel heard that Warburg
was looking for an assistant and recommended Krebs. This circle of influence allowed

Krebs to obtain his first paid position as an assistant to Otto Warburg at the Kaiser Wilhelm
Institut für Biologie at Berlin-Dahlem (see Figure 1). This stint was to change Krebs from a
physician to a biochemist. Of Warburg, Krebs wrote (1981):

Otto Warburg was the most remarkable person I have ever been closely associated with.
Remarkable as a scientific genius of the highest caliber, as a highly independent, penetrating
thinker, as an eccentric who shaped his life with determination and without fear, according to his
own ideas and ideals.

The Kaiser Wilhelm Institute was the leading scientific research facility of its time. In addition to
Warburg, its faculty included Emil Fischer, Richard Willstater, Max von Laue, Fritz Haber, Otto
Hahn, Lisa Meitner, and Michael Polanyl. Otto Meyerhof, a former student of Warburg, was
working in the same building and elucidated the glycolytic pathway responsible for the breakdown
of glucose to lactate. Students of biochemistry who passed through Kaiser Wilhelm included,
among others, Fritz Lipmann and Severo Ochoa, both of whom were to play a critical role in
showing the role of coenzyme A in the TCA cycle. In addition to the discipline and rigor required
for scientific research, Krebs learned from Warburg the techniques he had developed:
use of tissue slices, spectrophotometry and manometry. These techniques, most notably
manometry, were fundamentally important in leading Krebs to formulate the concept of a
metabolic cycle, first the urea cycle and later the TCA cycle. It is important to recognize in
understanding Krebs' work that the Warburg dictum “methods are everything” was dominant.
There was no theoretical master plan. Only during his later work on the thermodynamics linking
of cellular redox and phosphorylation states would Krebs remark in relation to that work, “This
is the first time I ever thought you could predict in biology.” In his early work, Krebs was a
thoroughly empiricist. He could measure the rates of reaction. He added various potential
participants in a metabolic pathway, and if they increased the overall rate, then he concluded
they were part of the pathway. Krebs' training is of central importance to understand the cycle.

As his biographer Holmes has pointed out, “Hans Krebs took 31 years to become a well-trained
independent scientific investigator, and only 9 months to make one of the most significant
discoveries of his generation in his chosen field.”
Disorders of Pyruvate Metabolism and Tricarboxylic Acid Cycle
Disorders of Tricarboxylic Acid Cycle
Tricarboxylic acid (TCA) cycle is the main electron donor of the ETC. In addition, succinate
dehydrogenase (SDH), which catalyzes the conversion of succinate to fumarate in the TCA cycle,
is also part of ETC complex II (Fig. 15.2). Disorders of TCA cycle mimic OXPHOS defects.
Table 15.2 enlists inborn errors of TCA cycle. Some TCA cycle defects are also implicated in
cancer pathogenesis as accumulated TCA cycle intermediates act as oncometabolites. Germline
mutations in genes encoding for TCA cycle enzymes have been associated with inherited cancer
predispositions (Table 15.3).

Disease Pathophysiology Gene Clinical

Presentation

α In KDH KDH is a multiplex enzyme Patients present in

Ketoglutarate deficiency, the made up of three units: E1, infantile or early E2, dehydrogenase
conversion of α- and E3. E1 and E2 are childhood period. specific (KDH) ketoglutarate to
for KDH. E3 They present with component is the
succinyl CoA is
deficiency same as in global delays,
impaired
PDH. E1 is encoded hypotonia, ataxia, by
OGDH. and seizures. There
are lactic acidosis
and high
αketoglutaric acid in
urine.

In SS deficiency, SS is made up of α and β Infantile lactic

Succinyl CoA
synthetase the conversion of subunits. acidosis,
(SS)/Succinyl succinyl CoA to SUCLG1 and SUCLA2 code encephalomyopathy,

Disease Pathophysiology Gene Clinical

Presentation
CoA ligase succinate is for α and β subunits, impaired. In hypotonia,
deficiency respectively. methylmalonic

addition, there is aciduria. Brain MRI

an accumulation may show lesions

of methylmalonic seen in Leigh

acid due to syndrome.

inhibition of the
Mitochondrial DNA
conversion of depletion can be
methylmalonyl estimated from the
CoA to succinyl muscle biopsy
specimen.
CoA. SS
deficiency also
causes disruption
of mitochondrial
nucleoside
diphosphate kinase
activity resulting
in

mtDNA
depletion.

Succinate SDH catalyzes SDHA, SDHB, SDHC,

dehydrogenase the conversion of and SDHD code for A, B,
(SDH) succinate to C, and D subunits, deficiency fumarate.
SDH deficiency
It is respectively.
presents with
encephalomyopathy
made up of four
resembling Leigh
subunits: A, B, syndrome. In
C, and D. addition, the
presentation could
Subunits A and B be cardiomyopathy,
are also part of skeletal myopathy,
complex II. FAD exercise intolerance,

Disease Pathophysiology Gene Clinical

Presentation
is converted to ataxia, or optic FADH2 by the atrophy.
SDH reaction.

An electron from
FADH2 is
transferred to
Coenzyme Q by
complex II.
Fumarase FH
deficiency
In fumarase
deficiency, the
conversion of Most patients
fumarate to is present in infancy
malate with global
impaired. developmental
delays, hypotonia,
and seizures. In
addition,
dysmorphic features
consisting of frontal
bossing, depressed
nasal bridge, and
hypertelorism may
be seen.

Neuroimaging may
show agenesis of
corpus callosum,
hydrocephalus,
cortical

malformations, and
choroid plexus
cysts.

Cancer TCA Cycle Gene Pathophysiology

Predisposition Enzyme
 Syndrome

Hereditary Succinate SDHA, SDHB, SDHC, Heterozygous paraganglioma and

dehydrogenase SDHD, SDHAF2. germline

pheochromocytomas (SDH) SDHA, SDHB, SDHC, mutations in and

SDHD code for A, B, SDH genes are C,
and D subunits of SDH, inherited in
respectively. SDHAF2 code autosomal

for complex II assembly dominant

factor. fashion. Loss of
heterozygosity
leads to complete
loss of protein in
the affected cells.
The
accumulation of
succinate leads to
stabilization of
hypoxiainducible
transcription
factor-1α
(HIF1α) leading
to metabolic
reprogramming
of cell that
promotes
carcinogenesis.

Fumarase FH
Multiple cutaneous (Fumarate Heterozygous and uterine hydratase) germline
leiomyomas mutation in
(MCUL) FH is

Cancer TCA Cycle Gene Pathophysiology

Predisposition Enzyme Syndrome

autosomal
Hereditary inherited in leiomyomatosis and
dominant
renal cell cancer
fashion. Loss
(HLRCC) of
heterozygosity
leads to complete
loss of protein in
the affected cells.
The accumulation
of fumarate leads
to stabilization of
hypoxiainducible
transcription
factor-1α (HIF-

1α) leading to
metabolic
reprogramming of
cell that promotes
carcinogenesis

Leaks in the Tricarboxylic Acid (TCA) Cycle

Abstract:
The TCA cycle is an amphibolic pathway. Anaplerotic reactions replenish TCA cycle
intermediates when they leak away from the cycle. Oxaloacetate leaks away from the TCA
cycle to form pyrimidines and glucose. Succinyl-CoA leaks away from the TCA cycle to form
the porphyrins (including heme). Several intermediates may leak away from the TCA cycle to
form proteins. Citrate may leak away from the TCA cycle to reform acetyl-CoA and OAA in the
cytoplasm, with the former entering into lipid biosynthesis. Leaks in the TCA cycle of skeletal
muscle fibers are largely replenished through glucose oxidation. Amino acids may be used to
replenish leaks in the TCA cycle.
Tricarboxylic Acid (TCA) Cycle
Abstract:
Genetic abnormalities in TCA cycle enzymes are generally incompatible with life.
Isocitrate dehydrogenase catalyzes the rate-limiting reaction in the TCA cycle. Oxidation of 1
pyruvate molecule through the TCA cycle can yield 15 ATP equivalents. Although oxaloacetate
and acetyl-CoA are impermeable to mitochondrial membranes, citrate and other intermediates of
the cycle are permeable. Certain rodenticides can interrupt the TCA cycle. Several B vitamins are
required as cofactors in the TCA cycle. GTP generated in the TCA cycle can be used to drive the
conversion of OAA to PEP in hepatic gluconeogenesis.

Deregulation of the Cellular Energetics of Cancer Cells

Glutamine Metabolism
Glutamine fuels the TCA cycle in cancer
The TCA cycle, also known as the citric acid cycle or Krebs cycle, occurs in the mitochondria and
provides large amounts of energy in aerobic conditions by donating electrons to three NADH and
one FADH (flavin adenine dinucleotide), which donate electrons to the electron transport chain,
creating the proton gradient needed to drive ATP synthesis. The TCA cycle can also provide
intermediates for macromolecule synthesis including citrate for lipid synthesis, oxaloacetate for
aspartate production, and malate for NADPH production. In the TCA cycle, aconitase converts
citrate (which can be derived from pyruvate and oxaloacetate) to isocitrate, which is converted to
α-ketoglutarate by isocitrate dehydrogenase. In a multistep process, α-ketoglutarate is converted
to succinyl-CoA, then succinate, followed by fumarate, malate, and then oxaloacetate. In
quiescent cells, a large fraction of the TCA cycle substrate is glucose-derived. However, the
diversion of pyruvate to lactate in cancer cells leaves them dependent on alternative sources of
carbon to fuel the TCA cycle. Many cancer cells become dependent on the amino acid glutamine
to fuel the TCA cycle. Glutamine is by far the most abundant amino acid that circulates in the
bloodstream, and tumors import high levels of glutamine. Importantly, many cancer cells fail to
grow when glutamine-deprived in culture. Glutamine-derived carbons can enter the TCA cycle
after glutamine is converted to glutamate by the enzyme glutaminase (GLS), and then, glutamate
is converted to the TCA cycle intermediate α-ketoglutarate by the enzyme glutamate
dehydrogenase (GLUD; Figure 1) or the transaminases.

Glutamine plays many important roles in cancer metabolism, in both a TCA cycle-dependent and
TCA cycle-independent manner. By supplying α-ketoglutarate to the TCA cycle, glutamine is
capable of replacing the TCA cycle intermediates malate and oxaloacetate. Malate can be used to
produce NADPH through its conversion to pyruvate by malic enzyme (ME), which is critical for
providing the reducing power for building block synthesis. Oxaloacetate can be converted to
aspartate, which plays a key role in nucleotide synthesis and amino acid synthesis. In addition to
feeding into the TCA cycle, glutamine that is converted to glutamate can make TCA
cycleindependent contributions to metabolism. Glutamine-derived glutamate is used for the
synthesis of the amino acids proline and arginine. Glutamate is also central to the synthesis of the
antioxidant glutathione, which plays a key role in managing oxidative stress in cancer.
Role of Glutamate Transport in Alcohol Withdrawal
Key Facts on the Tricarboxylic Acid Cycle
The TCA cycle is also known as the Krebs Cycle (named for its discoverer, Hans Adolf
Krebs) and the citric acid cycle (named after the intermediate citric acid, or citrate).

The TCA cycle metabolizes acetate derived from carbohydrates, proteins, and fats to form
adenosine triphosphate (ATP), the body’s energy currency.

Oxidation of one glucose molecule through the combined action of glycolysis, the TCA
cycle, and oxidative phosphorylation yields an estimated 30–38 ATP molecules. TCA
cycle intermediates oxaloacetate and α-ketoglutarate give rise to the amino acids
aspartate and glutamate, respectively—both of which act as excitatory
neurotransmitters in the brain.

There are eight enzymes in the TCA cycle that oxidize acetyl-coenzyme A (acetyl-CoA)
into two molecules of carbon dioxide.

Citrate produced in the TCA cycle can also be used in fatty acid synthesis.

Recent Advancements on the Role and Analysis of Volatile

Compounds (VOCs) from Trichoderma
Tricarboxylic Acid Cycle
The tricarboxylic acid (TCA) cycle completes the oxidation of glucose to carbon dioxide in the
cell and provides an intermediate for the biosynthesis of amino acids and of some VOCs. Succinic
acid intermediate from the TCA cycle has been isolated from Trichoderma pseudokoningii
together with itaconic acid (Kamal et al., 1971). This compound has been obtained from the
decarboxylation of cis-aconitic acid, another intermediate in the TCA cycle. The imide pencolide
is produced by Trichoderma album (Ren, 1977), could be generated from the condensation of
citraconic acid (Z-2-methyl-2butene dioic acid) and the amino acid threonine. Carolic acid is
isolated from a Trichoderma sp. and it is shown to subsist as a mixture of E and Z-isomers in
solution (Turner and Aldridge, 1983).

Energy Metabolism
Energy Metabolism of Other Nutrients
The TCA cycle and pyruvate are central in the metabolism of carbohydrate, fat, and protein in the
fed and fasting state. Pyruvate can have three main fates depending on the metabolic
circumstances. It can be a substrate for gluconeogenesis, or it can undergo oxidative
decarboxylation to acetyl-CoA and either enter the TCA cycle, or be used for fatty acid
synthesis.Acetyl-CoA can be made from carbohydrates via pyruvate, from fatty acids via
βoxidation in the mitochondrial matrix or from the proteolysis of proteins to amino acids, some of
which are converted to acetyl-CoA.There are three main stores of metabolic fuels:
Triacylglycerols in adipose tissue, glycogen as a carbohydrate reserve in liver and muscle and
protein as a source of amino acids which can be oxidized via the TCA cycle or used as a substrate
for gluconeogenesis.