CHAPTER 2

METABOLISM

BNN 30104 BIOCHEMISTRY AND

BIOMOLECULAR TECHNIQUES

DR SITY AISHAH MANSUR

CORI CYCLE /
LACTIC ACID
CYCLE
 Definition of Cori Cycle

• The phases in the metabolism of carbohydrate:

1) Glycogenolysis in the liver
2) Passage of glucose into the circulation
3) Deposition of glucose in the muscles as glycogen
4) Glycogenolysis during muscular activity and conversion
to lactate, which is converted to glycogen in the liver.

• Also called lactic acid cycle

• Simple explanation:
Cori cycle - the phases in the metabolism of carbohydrate.
 How does the reaction take place
- In cell -
MUSCULAR ACTIVITY AND THE CORI CYCLE:
 The explanation:

1. Epinephrine binds to a receptor on the muscle cell membrane and

stimulates adenyl cyclase in the membrane.

2. Adenyl cyclase in the membrane catalyzes the formation of cyclic

AMP from ATP.

3. The increase of cyclic AMP activates a protein kinase. The binding of

cyclic AMP to an enzyme is an allosteric control where the enzyme is
"switched on" for activity.

4. The protein kinase causes phosphorylations (addition of phosphate)

on a series of phosphorylation enzymes which activates them to
finally produce glucose-1-phosphate.

4a. At the same time that enzymes are being activated for glycogen
breakdown, glycogen synthetase enzyme must be inactivated.
Glycogenesis must be "switched off" and glycogenolysis "switched
on."
5. Glucose-6-phosphate is the final result of the
initial stimulation by epinephrine or other
hormones such as glucagon. If this happened to a
muscle cell, then the glycolysis pathway is the next
step in the sequence. If this happened to a liver cell
stimulated by glucagon, then glucose is produced to
enter the blood stream.
 Cori cycle (Lactic acid cycle)
 Discoverers = Carl Ferdinand Cori and Gerty Cori
 Biochemical pathway that recycles lactate
produced by muscle during anaerobic
glycolysis. The lactate is released to the blood,
taken up by the liver & converted back to glucose,
which is released again to be used by muscle.

 New glucose can either be passed back to muscles

via the bloodstream to serve as an energy source, or
be stored in the liver as glycogen
 EXPLANATION

 Muscular activity requires ATP, which is provided by the breakdown

of glycogen in the skeletal muscles. The breakdown of glycogen, a process
known as glycogenolysis, releases glucose in the form of glucose-1-
phosphate (G-1-P).
 The G-1-P is converted to G-6-P by the enzyme phosphoglucomutase.
 G-6-P is readily fed into glycolysis, a process that provides ATP to the muscle
cells as an energy source. During muscular activity, the store of ATP needs to be
constantly replenished.
 When the supply of oxygen is sufficient, this energy comes from
feeding pyruvate, one product of glycolysis, into the Krebs cycle.
 When oxygen supply is insufficient, typically during intense muscular activity,
energy must be released through anaerobic metabolism.
 Lactic acid fermentation converts pyruvate to lactate by lactate dehydrogenase.
 Most important, fermentation regenerates NAD+, maintaining the
NAD+ concentration so that additional glycolysis reactions can occur. The
fermentation step oxidizes the NADH produced by glycolysis back to NAD+,
transferring two electrons from NADHto reduce pyruvate into lactate.
 At first glycolysis produces pyruvic acid which is then converted into acetyl
CoA and is metabolized in the citric acid cycle to make ATP using the
electron transport chain.
 If muscular activity continues, the availability of oxygen for use at the end
of the electron transport chain becomes the limiting factor and the cells
soon exhaust their supplies of oxygen. When this happens, the citric acid
cycle is inhibited and causes pyruvic acid to accumulate.
 However, glycolysis continues even under anaerobic conditions even
though the citric acid cycle works only under aerobic conditions.
Epinephrine at (1) stimulates the enzymes to work on glycogen.
Glycogenolysis at (2) is stimulated to make more glucose-6-phosphate.
 When the cells become anaerobic, glycolysis (3) continues if pyruvic acid is
converted to lactic acid (4). Remember that the synthesis of lactic acid
requires NADH from Step 5 in glycolysis and produces NAD+ so that Step 5
can continue.
 The formation of lactic acid buys time and shifts part of the metabolic
burden to the liver.
 What happen when lactic acid accumulate?

 Even though not as much ATP can be furnished by glycolysis alone, it is a significant
source of ATP when muscular activity continues for any length of time. The final
limiting factor in continued muscular activity is the build up of lactic acid. The lactic
acid eventually produces muscular pain and cramps which force discontinuation of
activity. Usually before this happens and after activity has ceased, lactic acid diffuses
out of the muscle cells and into the blood where it enters the liver.

 The body is very efficient in that lactic acid is sent in the blood (5) to the liver which
can convert it back to pyruvic acid (6) and then to glucose through
gluconeogenesis (8). The glucose can enter the blood (9) and be carried to muscles
and immediately used. If by this time the muscles have ceased activity, the glucose
can be used to rebuild supplies of glycogen through glycogenesis (10)

 This recycling of lactic acid is referred to as the Cori Cycle. The Cori cycle also
operates more efficiently when the muscular activity has stopped. At this time the
oxygen debt can be made up so that the citric cycle and electron transport chain also
begin to function again. In order for most of the lactic acid to be converted to glucose,
some must be converted to pyruvic acid and then to acetyl CoA (7). The citric acid
cycle and electron transport chain must provide ATP to "fuel" the gluconeogenesis of
the remainder of the lactic acid to glucose.
 Significance

 Prevention of lactic acidosis in the muscle under anaerobic conditions.

However, normally before this happens the lactic acid is moved out of the
muscles and into the liver.

 The cycle is also important in producing ATP, an energy source, during

muscle activity. The Cori cycle functions more efficiently when muscle
activity has ceased. This allows the oxygen debt to be repaid such that the
Krebs cycle and electron transport chain can produce energy at peak
efficiency.

 The drug metformin can cause lactic acidosis in patients with renal failure
because metformin inhibits the hepatic gluconeogenesis of the Cori cycle,
particularly the mitochondrial respiratory chain complex 1. The build up of
lactate and its substrates for lactate production, pyruvate and alanine, lead
to excess lactate. Normally, the excess lactate would be cleared by the
kidneys, but in patients with renal failure, the kidneys cannot handle the
excess lactic acid.
 LIPID
METABOLISM
 Fatty Acid Degradation

Citric
Fatty Acetyl-
acid
acid CoA
cycle

Fatty acid degradation is the process in which fatty acids are broken down
into their metabolites, in the end generating acetyl-CoA, the entry molecule
for the citric acid cycle, the main energy supply of animals.
Myocardial metabolism. CPT, carnitine-palmitoyl-transferase; IMM, inner
mitochondrial membrane; OMM, outer mitochondrial membrane; PDH, pyruvate
dehydrogenase. Free-fatty-acid oxidative pathway in red. Glucose oxidative
pathway in blue.
 Liver Cell

FAT CELL Glycerol Glycolysis

Glycolysis Pyruvate

Triacylglycerol Gluconeogenesis
Glucon eogenesis Glucose
Other Tissues
Fatty
Acids Fatty acid oxidation Acetyl-CoA

CAC
In blood via
serum
albumin

CO2 + H2O
MOBILIZATION OF LIPIDS

DEGRADATION OF
TRIGLYCERIDES TO
FATTY ACIDS AND
GLYCEROL
LIPASE HYDROLYSIS OF TGAs
 Perilipin: mobilization of TAGs (precursor), and release of
ATGL co-activator (CA).
 ATGL: Adipose triglyceride lipase. Hydrolysis of one FA
from TAG to produce DAG
 + FA.
 HS-lipase: Hormone sensitive lipase. Hydrolysis of FA
from DAG to produce MAG +
 FA.
 MAG lipase: Hydrolysis of remaining FA from MAG to
produce glycerol and FA.

 The FA is then activated into Acyl-CoA form

 Fatty acid degradation is the process In which fatty acids
are broken down into
their metabolites.
 This process generates acetyl-CoA, which is the entry
molecule of the citric
acid cycle, the main energy supply of animals.

FA degradation

Lipolysis
of and
release Activation β-
from and oxidatio
adipose transport n
tissue into
mitochon
dria
Lipolysis of and release from adipose tissue
 In the process of degradation, FA are stored in fat cells.
 The breakdown of this fat is known as LIPOLYSIS.
 The products of lipolysis, free fatty acids are released into the bloodstream
and circulate throughout the body
 During the breakdown of triacylglycerols into fatty acids, more than 75%
of the fatty acids are reconverted to triacylglycerol even in cases of
starvation and exercise.

The enzymes of Lipolysis :

Hormone-Sensitive
Lipase (catalyzes
intracellular lipolysis)
Lipoprotein Lipase
(catalyzes hydrolysis of
circulating
triacylglycerols)
ACTIVATION AND TRANSPORT INTO
MITOCHONDRIA 1. Free fatty acids bound to albumin
are released and delivered via the
blood to tissues
2. The lipoprotein system also
delivers fatty acids to the liver and
other tissues
3. Fatty acids within the cell bound to
fatty acid binding protein
4. Fats may also be derived by
synthesis (lipogenesis) or
breakdown of triacylglycerols or
phospholipids. These fats are then
activated to their acyl CoA form by
acyl CoA syntheses (ACS)
5. Once in an "acyl CoA" form it is
converted into a carnitine
derivative for transport into the
mitochondria.
6. It is then subjected to β-oxidation
7. Acetyl CoA produced by β-
oxidation feeds into the citric acid
cycle for energy production
1. The product of acyl CoA synthetase,
(ACS) long-chain acyl CoA, cannot Mitochondrial Uptake and b-Oxidation of
pass through the inner Fatty Acids
mitochondrial membrane.
2. So, it is transformed by carnitine
palmitoyl transferase I (CPT-I) to
acylcarnitine. (-) Malonyl CoA
(lipogenesis)
3. Carnitine-acylcarnitine translocase
acts as a membrane carnitine
"exchange transporter".
4. Acylcarnitine goes in and a
carnitine comes out.

- The acylcarnitine reacts with CoA

via carnitine palmitoyl
transferase II (CPT-II), attached to
the inner membrane.
- Acyl CoA is reformed in the
mitochondrial matrix and
carnitine is liberated.
- Deficiencies in CPT's lead to
considerable muscle weakness,
as fatty acids are a major fuel for
muscles.
 β-Oxidation
Once inside the mitochondria, the β-oxidation of
fatty acids occurs via five recurring steps:
 Activation by ATP
 Oxidation by FAD,
 Hydration,
 Oxidation by NAD+,
 Thiolysis,
 The final product is acetyl-CoA, the entry
molecule for the citric acid cycle.
What if the FA do not have an even no. of carbons or
not fully saturated??
 Use different mechanisms
ODD CHAIN FATTY ACIDS

STEPS :
1. Propionyl-CoA carboxylated using a bicarbonate ion into D-methylmalonyl-
CoA in a rxn involves a biotin co-factor, ATP and the enzyme propionyl-CoA
carboxylase.
2. D-methylmalonyl-CoA converted into L-methylmalonyl CoA by
methylmalonyl-CoA epimerase.
3. L-methylmalonyl-CoA undergoes intracellular rearrangement, which is
catalysed by methylmalonyl-CoA mutase to form succiniyl-CoA.
4. Succinyl-CoA CONVERT INTO ACETYL COA… and then enter CA cyle.
• Also catabolized by B-oxidation, BUT 2 additional enzymes
(isomerase & novel reductase) required to handle the cis double bonds
EXAMPLE:
 PENTOSE
PHOSPHATE
PATHWAY
 What is…

 A metabolic pathway parallel to glycolysis that generates NADPH

and pentoses (5-carbon sugars) as well as Ribose 5-phosphate.

 Does oxidize glucose and under certain condition can completely

oxidize glucose to CO₂ and water.
 Primary function for this pathway

 To generate reducing equivalent in form of NADPH for reductive

biosynthesis reaction

 To provide the cell with ribo-5-phosphate (R5P) for the synthesis

of nucleotides and nucleic acids

 Can metabolize dietary pentose sugar derived from the digestion

of nucleic acid
There are two distinct phases in the pathway.

 Oxidative phase, in which NADPH is generated

 Non-oxidative synthesis of 5-carbon sugars

For most organisms, the pentose phosphate pathway takes place in the
cytosol; in plants, most steps take place in plastids
 As RBCs age, enzyme activities involved in glucose metabolism
diminish, including G6PD, reducing energy production and ability to
protect cell membrane integrity and hemoglobin from oxidation
 The reactions of fatty acid biosynthesis and steroid biosynthesis
utilize large amounts of NADPH.
 As a consequence, cells of the liver, adipose tissue, adrenal
cortex, testis and lactating mammary gland have high levels of
the PPP enzymes.
 In fact 30% of the oxidation of glucose in the liver occurs via the
PPP.
 The only source of NADPH in RBCs is via the PPP, in which G6PD is
the rate-limiting step
 Additionally, erythrocytes utilize the reactions of the PPP to generate
large amounts of NADPH used in the reduction of glutathione.
 PRECURSORS
 Glucose 6-Phosphate, Glu 6-P, which comes
from other pathways, glycolysis.

 Glucose 6-phosphate dehydrogenase, and one

important cofactor, NADP+.

 Enzyme lactonase
 PATHWAY
 It starts with Glucose 6-Phosphate, Glu 6-P, which comes from
other pathways, glycolysis for example.

 Instead of using the enzyme glucose 6-phosphate isomerase to

isomerize the original reactant into fructose 6-phosphate for
glycolysis, cells use another enzyme, glucose 6-phosphate
dehydrogenase, and one important cofactor, NADP+, to oxidize
the Glu 6-P into 6-phosphoglucono-δ-lactone with NADP+
being reduced to NADPH.

 Next, the enzyme lactonase hydrolyzes 6-phosphoglucono-δ-

lactone into 6-phosphogluconate.

 In this step, cofactor NADP+ is used again as an oxidizing

agent to oxidize
 6-phosphogluconate to ribulose 5-phosphate in a
reaction catalyzed by the enzyme 6-phosphogluconate
dehydrogenase with the reduced NADPH as another
product and the release of carbon dioxide.

 Note that in every step of the pathway, an addition of

Magnesium cation helps stabilizing the reactions, which
involves releases of electrons and protons.

 In a ketose-aldose reaction catalyzed by the enzyme

phosphopentose isomerase, ribulose 5-phosphate is
isomerized into ribose 5-phosphate, a precursor for later
important reactions, such as DNA synthesis.
 PRODUCTS

The primary results of the pathway are:

 The generation of reducing equivalents, in the form of NADPH, used
in reductive biosynthesis reactions within cells (e.g. fatty acid
synthesis).

 Production of ribose 5-phosphate (R5P), used in the synthesis

of nucleotides and nucleic acids.

 Production of erythrose 4-phosphate (E4P) used in the synthesis

of aromatic amino acids.
SUMMARY OF PPP
GLUCOGENIC
AMINO
ACIDS
 DEFINITION
A glucogenic amino acid are amino acids that can be
converted into glucose through gluconeogenesis.

Each of the 20 amino acids has a separate catabolic pathway,

yet all 20 pathways converge into 5 intermediates, all of
which can enter the citric acid cycle. From the citric acid cycle
the carbon skeletons can be completely oxidized into CO2 or
diverted into gluconeogensis or ketogenesis.

 Glucogenic amino acids are broken down into one of the

following metabolites: pyruvate, α-ketoglutarate, succinyl
CoA, fumarate or oxaloacetate

 Larger amino acids, tryptophan, phenylalanine, tyrosine,

isoleucine andthreonine are both glucogenic and ketogenic
Glucogenic Amino Acids
 REACTION LOCATION IN THE CELL
 The pathway may begin in the mitochondria or
cytoplasm, this being dependent on the substrate
being used. Many of the reactions are the reversible
steps found in glycolysis. Glucogenic Amino Acid
Metabolism begins in the mitochondria with the
formation of oxaloacetate by the carboxylation of
pyruvate.

 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.
 REACTION LOCATION IN THE BODY
 The process takes place mainly in the liver and, to a
lesser extent, in the cortex of the kidneys.
 In many other animals, the process occurs during
periods of fasting, starvation, low-carbohydrate diets,
or intense exercise.
 Kidneys are the second most important site of this
reaction. Its main substrate is the carbon skeleton of
amino acids.
 The formation of oxaloacetate from pyruvate and TCA
cycle intermediates is restricted to the mitochondrion,
and the enzymes that convert Phosphoenolpyruvic
acid(PEP) to glucose are found in the cytos.
The Precursors
 Each intermediate uses different
types of glucogenic amino acid itself.

 All amino acids except lysine and

leucine are at least partly glucogenic.

 They are also had been divided into

two different types which are non-
essential and essential amino acids.
 Amino acids that form pyruvate
(Alanine, Serine, Glycine, Cystine, Threonine)
1) Alanine

Alanine loses its amino group by transamination to form

pyruvate

2) Serine and 3)
Glycine Inter conversion of serine and
glycine
Serine is be converted to pyruvate by serine dehydratase.

4) Cystine

5)
Threonine
 Amino acids that form α-ketoglutarate
(Glutamine, Proline, Arginine, Histidine)
1) Glutamine: Oxidative
deamination

oxidative deamination by
glutamine dehydrogenase Α-ketoglutarate
2) Proline:It is oxidized to glutamate. Glutamate is then oxidatively deaminated to form α-
ketoglutarate

3) Arginine: This aa is cleaved by arginase to produce ornithine. Ornithineis subsequently

converted to: α-ketoglutarate
4) Histidine
Amino acids that form oxaloacetate
(Aspargine and Aspartate)

Asparagineis hydrolyzed by Asparaginase, liberating

ammonia and Aspartate

Aspartate loses its amino group by transamination to form

oxaloacetate

condenses with acetyl

CoA to form citrate in Glucogenic
the first reaction of the
Krebs cycle
 Amino Acids that form fumarate
(Phenylalanine and Tyrosine)

1) Phenylalanine and
2) Tyrosine

Hence these two aa are both

glucogenic and ketogenic
 Amino acids that form succinyl CoA
(Methionine, Valine, Isoleucine, Threonine)
Methionine
1) Methionine condenses with ATP to form S-adenosylmethionine
2) Methyl group is activated and transferred to oxygen, nitrogen or carbon
atoms.
3) The reaction product is S-adenosylhomocysteine
4) S-adenosylhomocysteineis hydrolyzed to homocysteine

Homocysteine has two fates:

a) In case of methionine deficiency it is remethylated to methionine
b) If methionine stores are adequate, it enters transulferation pathway to form
cysteine and α-ketobutyrate, which is oxidatively decarboxylated to form
propionyl CoA which is then converted to Succinyl CoA.
Amino acids that form succinyl CoA
(Valine, Isoleucine and Threonine)
KETOGENIC
AMINO
ACIDS
 KETOGENIC AMINO ACID
METABOLISM
• A ketogenic amino acid is an amino acid that can be degraded
directly into acetyl-CoA, which is the precursor of ketone bodies.
• Ketogenic amino acids are unable to be converted to glucose as both
carbon atoms in the ketone body are ultimately degraded to carbon
dioxide in the citric acid cycle.
• Ketogenesis is the biochemical process by which organisms produce
a group of substances collectively known as ketone bodies by the
breakdown of fatty acids and ketogenic amino acids.

• The three ketone bodies are acetone, acetoacetic acid and beta-
hydroxybutyric acid.
 In humans, two
amino acids are
exclusively
ketogenic:
• Leucine
• Lysine

 In humans, five
amino acids are
both ketogenic and
glucogenic:
• Isoleucine
• Phenylalani
ne
• Threonine
• Tryptophan
• Tyrosine
• Ketone bodies are produced mainly in the mitochondria of
liver cells.

• Synthesis can occur in response to an unavailability of

blood glucose, such as during fasting.

• Ketogenesis takes place in the setting of low glucose

levels in the blood, after exhaustion of other cellular
carbohydrate stores, such as glycogen.

• It can also take place when there is insufficient insulin

(e.g. in diabetes), particularly during periods of
"ketogenic stress" such as intercurrent illness.
• The three ketone bodies, each synthesized from acetyl-CoA molecules, are:
i. Acetoacetate, which can be converted by the liver into β-hydroxybutyrate,
or spontaneously turn into acetone.

ii. Acetone, which is generated through the decarboxylation of acetoacetate,

either spontaneously or through the enzyme acetoacetate decarboxylase. It
can then be further metabolized either by CYP2E1 into hydroxyacetone
(acetol) and then via propylene glycol to pyruvate, lactate and acetate (usable
for energy) and propionaldehyde, or via methylglyoxal to pyruvate and
lactate.

iii. β-hydroxybutyrate (not technically a ketone according to IUPAC

nomenclature) is generated through the action of the enzyme D-β-
hydroxybutyrate dehydrogenase on acetoacetate.
Overview of Energy Metabolism

Figure 8.14
 Quick Review
 Acetyl CoA is the gateway molecule for all energy
nutrients
 Acetyl CoA combines with oxaloacetate to form
citrate in the first step of the TCA cycle
 The TCA cycle produces
 Two energized coenzymes
 Two molecules of CO2
 Small amount of energy as GTP
 Electrons from hydrogen atoms in coenzymes enter
the electron transport chain
 Protons are used to form ATP during the electron
transport chain
Metabolism Adapts during Feasting or Fasting

Figure 8.15
 Quick Review
 Ingestion of excess kilocalories
 Anabolic metabolism

 Excess energy from carbohydrates, proteins, fats, and

alcohol are converted to fat and stored

 Fasting or starvation
 Catabolic metabolism

 Fat is broken down to fatty acids

 Glycerol and amino acids maintain blood glucose

 Lack of glucose leads to formation of ketone bodies for

energy