The Citric Acid Cycle

Kreb’s Cycle
TCA (tricarboxylic acid cycle)

Reference Text
Timberlake, K. (2009). Chemistry: An introduction to general, organic, and biological chemistry.
10th edn. Pearson Prentice Hall, New Jersey.
 The three fates of pyruvate
 Pentose phosphate pathway
 Introduction to the Krebs Cycle

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 Three reactions are possible for pyruvate
depending on the availability of oxygen.
1. If oxygen is available ATP is produced and this
reaction is aerobic and occurs in the mitochondria.
2. In humans: if oxygen is NOT available : production of
lactic acid. This reaction is anaerobic and occurs in the
cytosol.
3. In bacteria and yeasts: if oxygen is NOT available:
production of alcohol.

What conditions would promote the metabolism of pyruvate

aerobically or anaerobically?

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1. Aerobic catabolism:
• If O2 is present, pyruvate enters the mitochondrion as acetyl-
CoA.

• For this reaction, thiamin as TPP (thiamine pyrophosphate),

niacin (as NAD+), and Coenzyme A are needed.

• Pyruvate, a 3-carbon molecule, is oxidised to Acetyl CoA, a

2-carbon molecule, and carbon dioxide. This reaction is
complex and involves different enzymes.

• Acetyl CoA can then enter the Kreb’s cycle (citric acid cycle)
to produce more ATPs.

• We will discuss this pathway more extensively later.

C3H3O3 + NAD+ + CoA  C2H3O-CoA + CO2 + NADH + H+

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2. Anaerobic catabolism (in humans)
– if O2 is not present, pyruvate is broken down to
lactate.

– This would happen during strenuous exercise

whereby oxygen is not delivered quickly enough
to the muscles.

– This process is referred to as “fermentation”.

– Cellular respiration may still be happening, but it

is not the major source of ATP while fermentation
is occurring.

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2. Anaerobic catabolism (cont.):
– This reaction requires NADH and hydrogen and
produces NAD+.

– Supply of NAD+ is limited, i.e. glycolysis can only

continue if NADH + H+ is relieved of its hydrogen.

 If oxygen is present NADH will bind to oxygen and

form water and ATP in the electron transport chain.
 If oxygen is not present the NADH + H+ will unload its
hydrogen and pass them onto pyruvate to form lactate.
 Either way NAD+ is regenerated and glycolysis can
continue.

C3H3O3 + NADH + H+  C3H5O3 + NAD+

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– Very little energy is gained in this pathway.

– Lactate is a “dead end” metabolically in the

immediate absence of oxygen.

– Only 2 ATPs have so far been made from glucose to

the production of lactate.

What is the point of making lactate then?

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– The formation of lactate buys time and shifts part of
the metabolic burden from the muscles to the liver.
– Lactate must be converted back to pyruvate before
it can be metabolised and this is done in the liver.
– Because cell membranes are permeable to pyruvate
and lactate, these substances can easily leave or be
absorbed by the cells.
– The liver converts pyruvate back into glucose and
sends it back to the blood to be used as an energy
source. This is referred to as gluconeogenesis.
– We will discuss this process in later lectures.

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– The muscle cells take up the glucose from the blood
and use it to make ATP.

HOW? HOW MANY ATPs?

– The only purpose to reduce pyruvate into lactate in

the muscle is to regenerate NAD+ needed for
glycolysis.
– Without the regeneration of NAD+, glycolysis cannot
happen and no ATP can be made.

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– If there is no oxygen, pyruvate is converted
to lactate.

– The liver changes lactate into glucose, which

is then picked up by the muscles to make
ATPs.

Why is lactate changed to pyruvate in the

liver and not in the muscles?

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  There are also fungi and bacteria that produce
lactic acid during fermentation.

 We use these  Similar kinds of

microorganisms to microbial fermentation
transform milk into turn soybeans into soy
cheese and sauce and cabbage
yoghurt which into sauerkraut.
explains the sour
flavour of these
foods.

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3. Alcohol formation:
 Often referred to as “fermentation”.
 This is the formation of alcohol from sugars by
yeast or micro-organisms under anaerobic
conditions which converts pyruvic acid to
ethanol.
Remember: in muscles pyruvic acid is converted to
lactic acid NOT ethanol!
 Pyruvate is first changed to acetaldehyde and
then to ethanol.
 This is the reverse reaction humans use when
breaking down alcohol using similar enzymes.

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 Yeast produces bread or alcoholic
beverages such as beer and wine.
 The carbon dioxide is what makes
champagne and beer bubbly.

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 The enzyme involved in the conversion of pyruvate to
acetaldehyde requires thiamine.

Pyruvate + H+  acetaldehyde + CO2

 The enzyme involved in the conversion of

acetaldehyde to ethanol requires niacin (NAD) and
zinc.

Acetaldehyde + NADH + H+ ethanol + NAD+

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  The conversion of glucose to ethanol is:

Glucose + 2 Pi + 2 ADP + 2 H+  2 ethanol + 2 CO2 + 2 ATP + 2 H2O + 2 NAD+

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ummm....what about
an alternative to
carbohydrate
oxidation?

or

Do we only burn
carbohydrates for
energy?

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 This pathway is an alternative to glycolysis and
is considered primarily as an anabolic pathway
(biosynthesis) in the cytosol of cells.
 It operates in varying extends in different cells
and tissues.
 Begins at glucose-6-phosphate (G6P)
 There are two distinct phases in the pathway
that:
 generates NADPH
 generates pentose sugars (5-carbon sugars).

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The generation of NADPH is critically
important in cells because it provides an
electron source for biosynthesis.
NOTE: NADP is identical to NAD except that
it is phosphorlyated.
Therefore, rapidly dividing cells have a great
demand on this system.
NADPH is required for:
- Fatty acid synthesis to occur (whiclogically
is also placed in the cytsol).
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 The PPP pathway also acts to siphon 6 carbon
moieties off from oxidation to be used to build
nucleotides needed for DNA synthesis.

There are some interesting

diseases associated with lack
Of NADPH generation due
to faulty PPP.

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  The PPP pathway also
provides a convenient
way to digest foods that
are high in nucleic acids
such as organ meats 

Lambs Fry...umm...

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 Sir Hans Adolf Krebs
was born at
Hildesheim, Germany,
on August 25th, 1900.
 Died in 1981
 Discovered the
metabolic pathway
which has been given
his name in his
honour.

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  The central reaction for all catabolic pathways;
degrades acetyl-CoA to CO2 and energy.
 Most fuel molecules (carbohydrates, lipids, and
proteins) enter the cycle as Acetyl-CoA.
 The citric acid cycle occurs in the mitochondria.
 Oxaloacetate (4-carbon molecule) combines with
Acetyl-CoA (2 carbon) to form a 6-carbon molecule.
 Note: oxaloacetate is essential in the running of the Kreb’s
cycle.
 Oxaloacetate and Acetyl CoA form citric acid (hence
the name of the cycle).

C2H3O-CoA + 3 NAD+ + FAD + ADP + Pi + 2 H2O

2 CO2 + 3 NADH + 2H+ + FADH2 + CoA + ATP
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 2 carbon atoms leave the cycle in the form of CO 2
 4 pairs of electrons (H+) leave the cycle (3 NADH and 1
FADH2).
 1 high-energy phosphate bond (in the form of GTP)
 2 water molecules are consumed.
 In the next step of metabolism the NAD and FADH 2
will be metabolised into ATP.
 1 NADH makes 3 ATPs
 1 FADH2 makes 2 ATPs

How many ATPs are formed?

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 There are a total of 5 co-enzymes required to
the conversion of pyruvate and operation of
the Krebs cycle:
 FAD
 Thiamine pyrophosphate
 Lipoic acid
 Co-enzyme A
 NAD

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 Many pathways exist from the Krebs cycle that
can draw carbons off to engage in biosynthetic
pathways.
 Examples include fatty acid synthesis, heme,
amino acids and purines.
 Literally, the TCA ensures that substrates are
present to make other molecules.

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