Biochemistry Notes Test 2 2023

THE UNIVERSITY OF ZAMBIA
SCHOOL OF MEDICINE
PHYSIOLOGICAL SCIENCES DEPARTMENT
NUTRITION AND FAT SOLUBLE VITAMINS
Mwale Nicholas K.(PhD)

mwalenicholask@unza.zm
Conference Room, Adult Extension Studies Building, Ridgeway Campus
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OUTLINE
OBJECTIVE of the topic:
• Describe normal nutrition and deviations from the normal state which can
lead to diseases.
• NUTRITION
– Definition
– Importance of Nutrition
• Importance of food related to death
• Factors that contribute to death
– Classification of Nutrients
– Why we eat what we eat?
– Dietary Reference Intakes
– Dietary carbohydrate
– Dietary fats
– Dietary proteins
– Nitrogen balance
– Fuel stores
• VITAMINS
– Overview
– Fat-soluble vitamins
• A,D,E,K
– Water-soluble vitamins
• Thiamine (Vitamin B1), Riboflavin (Vitamin B2), Pantothenic
acid (Vitamin B5), Niacin (Nicotinic acid B3), Pyridoxine (Vitamin
B6), Biotin (Vitamin B7), Cobalamin (Vitamin B12), Folic acid
(pteroylglutamic acid B9)
• Vitamin C
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NUTRITION
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3. CLASSIFICATION OF NUTRIENTS.
• Nutrients are carbohydrates, proteins, lipids, vitamins,
minerals and water.
Various Classifications:
TYPE 2: Classification
MACRONUTRIENTS: carbohydrates, proteins, lipids, and water (required in
larger amounts)
MICRONUTRIENTS: vitamins, minerals (required in lesser amounts)
ENERGY YEILDING NUTRIENTS/ENERGY CONTENT OF FOOD:
Carbo: 1g=4kcal Prot: 1g=4kcal Lipids: 1g=9kcal Alcohol: 1g=7kcal (not a nutrient,
is toxic)
What is the total energy in 10g of carb, 5g of protein and 2g of fats?4
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4. WHY WE EAT WHAT WE EAT?
Some:
1. Social factors -availability (wedding),
-economical (money),
-convenience (fast foods)
2. Behavioural factors -emotions(boredom, stress)
-value (religion)
-body image
3. Genetics? Find out what this is about

4. Personal preference - Taste (sweet vs salty food)
-Upbringing
-Habit
-View of food
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5. DIETARY REFERENCE INTAKES
(DRI).
Refers to different reference of the
intakes of nutrients:
1. Estimated Average Requirement (EAR)
-Average amount sufficient for half the
population (doesn’t meet the req.) Eg Joe
req. 200kcal, Biden req 300kcal; average-
250kcal ?????
2. Recommended Dietary Allowance (RDA)
-Meets the needs of 98% of population.
-Based on scientific evidence; Calculated
from double EAR.
3. Adequate Intake (AI)
-Doesn’t meet the needs as evidence is on-
going.Once conclusion made it becomes RDA
4. Tolerable Upper Intake Levels (UL)
-Maximum consumption before it becomes
toxic.
-This helps against over-consumption.
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DIETARY REFERENCE INTAKES cont...
6. Estimated Energy Requirement (EER)
-Amount of calories needed for daily function ie
-vital functions of body (breathing, digestion)
-physical activities (walking, exercises)
-This maintains energy balance (calories consumed =the energy
expended) in a healthy adult of a defined age, gender, and height
whose weight and level of physical activity are consistent with
good health.
NOTE
1. Differences in the genetics, body composition, metabolism, and
behavior of individuals make it difficult to accurately predict a
person’s caloric requirements.
2. As a general guide;
-very active adults require 40 kcal/kg/day to maintain body weight
-moderately active adults require 35 kcal/kg/day to maintain body
weight
-sedentary adults require about 30 kcal/kg/day to maintain body
weight
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Note cont...
3. To stay in energy balance, we must, on average, consume
an amount of food equal to our daily energy expenditure
(DEE).
-The daily energy expenditure (DEE) = the energy to
support our basal metabolism (basal metabolic rate
or resting metabolic rate) and our physical activity,
+ the energy required to process the food we eat
(diet-induced thermogenesis (DIT) or thermal
effect of food).
• It can be summarised thus:

Daily energy expenditure= RMR(or BMR) +Physical Activity
+DIT
where RMR is the resting metabolic rate;
BMR is basal metabolic rate;
DIT is diet-induced thermogenesis.
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6a) BASAL METABOLIC RATE OR RESTING METABOLIC RATE
• It is a measure of the energy required to carry out normal body
functioning of the lungs, kidneys and brain, the pumping of the
heart, the maintenance of ionic gradients across membranes, the
biochemical reaction pathways, maintenance of cellular integrity,
etc.
Factors Affecting BMR Expressed per kg Body Weight
1. Gender (males higher than females) 2.Body temperature
(increased with fever) 3.Environmental temperature (increased in
cold) 4. Thyroid status (increased in hyperthyroidism) 5. Pregnancy
and lactation (increased) 6. Age (decreases with age)
• A rough estimate of the BMR may be obtained by assuming it is 24

kcal/day/kg body weight and multiplying by the body weight.
• Eg For an average adult, the BMR is about 1,800 kcal for men (70
kg) and 1,300 kcal for women (50 kg). Using the above formula, this
70kg man’s RMR=1680kcal and the 50kg woman is 1200kcal.
• From 50–70% of the daily energy expenditure in sedentary
individuals is attributable to the RMR
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6b) DIET-INDUCED THEMOGENESIS/THERMIC EFFECT OF
FOOD:
• After the ingestion of food, our metabolic rate increases
because energy is required to digest, absorb, distribute,
and store nutrients.
• This can be observed by the production of heat by the
body which increases as much as 30% above the resting
level during the digestion and absorption of food.
NOTE:
• The energy required to process the types and quantities
of food in the typical American diet is probably equal to
approximately 10% of the kilocalories ingested.
• This amount is roughly equivalent to the error involved in
rounding off the caloric content of carbohydrate, fat, and
protein to 4, 9, and 4, kcal/g respectively and so thermic
effect of food is often ignored and calculations of the
daily energy expenditure are based simply on the RMR and
the energy required for physical activity. 10
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6c) PHYSICAL ACTIVITY:
• Physical muscular activity provides the greatest
variation in energy expenditure.
• Therefore the amount of energy consumed depends on
the duration and intensity of the physical activity.
• Estimation of daily expenditure of energy on physical
activity can be done by recording the type and duration of
all activities
• A rough estimate of the energy required per day for
physical activity can be made by using
• a value of 30-50% of the RMR (per day) for a very
sedentary person (such as a medical student who
does little but study) and
• a value of 60 to 70% of the RMR (per day) for a
person who engages in about 2 hours of moderate
exercise per day or
• a value of 100% or more of the RMR for a person
who
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does several hours of heavy exercise per day
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7 Acceptable Macronutrient Distribution Range (AMDR)
-How many calories from carbohydrates, proteins, lipids
are acceptable?
45-65% kcal for carbohydrates
20-35% kcal for lipids
10-35% kcal for proteins
-This range prevents chronic diseases
How does one know their AMDR?

By performing a Nutritional Assessment.
-Deficiency or excess overtime leads to malnutrition
-under nutrition
-over nutrition
Similar symptoms include diarrhea, skin rashes, fatigue
etc
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In the Nutritional Assessment, get
1. History (dietary and health)
2. Anthropometric measurements
-aims to achieve healthy body weight
WHAT IS THE IDEAL WEIGHT?
• This is guided by the body mass index (BMI), calculated
as weight/height2 (kg/m 2)
• Adults with BMI values below 18.5 are considered
underweight.
• Those with BMIs between 18.5 and 24.9 are considered
to be in the healthy weight range,
• Those between 25 and 29.9 are in the overweight or pre-
obese range
• Those above 30 are in the obese range.
3. Physical examination -eyes (discoloration/ pale)
-tongue
-skin
4. Laboratory blood test 13
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6. DIETARY CARBOHYDRATES
Classification of Carbohydrates
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DIETARY CARBOHYDRATES cont....
• No specific carbohydrates have been identified as
dietary requirements BUT their role is to provide energy
• They are not essential nutrients, because the carbon

skeletons of most amino acids can be converted into
glucose.
• Absence of dietary carbohydrate leads to ketone body

production, and degradation of body protein whose
constituent amino acids provide carbon skeletons for
gluconeogenesis.
• The RDA for carbohydrate is set at 130 g/day for

adults and children, based on the amount of glucose used
by carbohydrate-dependent tissues, such as the brain
and erythrocytes.
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7. DIETARY FATS
• Although most lipids required for cell structure, fuel
storage, or hormone synthesis can be synthesized from
carbohydrates or proteins, we need a minimal level of
certain dietary lipids for optimal health-known as
essential fatty acids.
• These are required in our diet because we cannot
synthesize fatty acids with these particular arrangements
of double bonds.
• The essential fatty acids α-linoleic and α-linolenic acid are
supplied by dietary plant oils, and eicosapentaenoic acid
(EPA) and docosahexaenoic acid (DHA) are supplied in fish
oils.
• They are the precursors of the eicosanoids (a set of
hormone-like molecules that are secreted by cells in small
quantities and have numerous important effects on
neighboring cells).
• The eicosanoids include the prostaglandins,
thromboxanes, leukotrienes, and other related
compounds. Dr. Mwale, 2023
DIETARY FATS cont…
A. Plasma Lipids
• Plasma cholesterol may arise from the diet (animal
products) or from endogenous biosynthesis and in either
case, cholesterol is transported between the tissues in
combination with protein and phospholipids as lipoproteins.
B. Classes of Lipoproteins
1. Low-density lipoprotein (LDL) 2. high-density lipoprotein
(HDL):
• The level of plasma cholesterol is not precisely regulated,
but rather varies in response to the diet and so elevated
levels of LDL result in an increased risk for Coronary
Heart Disease (CHD).
• This risk increases progressively with higher values for
serum total cholesterol and a much stronger correlation
exists between the levels of blood LDL cholesterol and
heart disease.
• Note that high levels of HDL cholesterol have been
associated with a decreased risk for heart disease.
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C. Dietary Fats-Triacyglycerols
• are quantitatively the most important class of dietary fats
where their influence on blood lipids is determined by the
chemical nature of their constituent fatty acids ie
saturated , monosaturated, polysaturated fats and trans-
fatty acids.
1. SATURATED FATS
• These are triacylglycerols composed primarily of fatty acids
whose hydrocarbon chains do not contain any double bonds.
• Saturated fatty acids with carbon chain lengths of 14
(myristic) and 16 (palmitic) are most potent in increasing the
serum cholesterol. Stearic acid (18 carbons—found in many
foods including chocolate) has little effect on blood
cholesterol.
• Consumption of saturated fats is positively associated with
high levels of total plasma cholesterol and LDL cholesterol,
and an increased risk of CHD.
• Other sources include are dairy and meat products and
some vegetable oils, such as coconut and palm oils.
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2. Monounsaturated fats
• Triacylglycerols containing primarily fatty acids with
one double bond
• Source include vegetables and fish.
• When substituted for saturated fatty acids in the
diet, monounsaturated fats lower both total plasma
cholesterol and LDL cholesterol, but maintain or
increase HDL cholesterol.
3. Polyunsaturated fats
• Contain primarily fatty acids with more than one
double bond
• The effects of polyunsaturated fatty acids (PUFAs)
on cardiovascular disease is influenced by the location
of the double bonds within the molecule.
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Polyunsaturated fats cont…
a) ω-6 Fatty acids
• The location of the first double bond beginning at the
sixth bond position when starting from the methyl end
of the fatty acid molecule are referred to as ω-6
Fatty acids
• Consumption of fats containing these long-chain ω-6
PUFAs, principally linoleic acid, 18:2(9,12), obtained
from vegetable oils, lowers plasma cholesterol when
substituted for saturated fats.
• Plasma LDL are lowered, but HDL, which protect
against CHD, are also lowered.
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Polyunsaturated fats cont…
b) ω-3 Fatty acids
• These are long-chain, PUFAs, with the first double
bond beginning at the third bond position from the
methyl end.
• Dietary ω-3 polyunsaturated fats suppress cardiac

arrhythmias, reduce serum triacylglycerols, decrease
the tendency for thrombosis, lower blood pressure,
and substantially reduce risk of cardiovascular
mortality,
• However, they have little effect on LDL or HDL
cholesterol levels.
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4. Trans fatty acids
• These are chemically classified as unsaturated fatty
acids, but behave more like saturated fatty acids in
the body, that is, they elevate serum LDL (but not
HDL), and they increase the risk of CHD.
• They do not occur naturally in plants, but occur in

small amounts in animals.
• They are formed during the hydrogenation of liquid

vegetable oils, for example, in the manufacture of
margarine and partially hydrogenated vegetable oil.
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SUMMARY OF THE ESSENTIAL FATTY ACIDS
TYPE OF FAT METABOLIC EFFECTS EFFECTS ON DISEASE
PREVENTION
Saturated Increases LDL Little effect on Increased incidence of
HDL CHD; may increase risk of
prostate and colon cancer
Monounsaturated Lowers LDL Lowers HDL Reduced incidence of CHD
Polysaturated ω-6 Lowers LDL Lowers HDL Reduced incidence of CHD
Provide arachidonic acid, an
important precursor of
prostaglandins and leukotrienes
Polysaturated ω-3 Little effect on Little effect on Reduced incidence of CHD
LDL HDL Reduced risk of sudden
Supresses cardiac arrhythmias, cardiac death
reduce serum triacyglycerols,
decrease the tendency for
thrombosis, lower blood pressure
Trans Increases LDL Lowers HDL Increased incidence of
CHD
8. DIETARY PROTEIN
• High quality of a dietary protein contain all the essential
amino acids required for tissue maintenance.
• Nine of the 20 amino acids needed for the synthesis of
body proteins are essential—that is, they cannot be
synthesized in humans and so must be obtained from the
diet.
A. 9 ESSENTIAL AMINO ACIDS

• These are lysine, isoleucine, leucine, threonine, valine,
tryptophan, phenylalanine, methionine, and histidine.
• Certain amino acids are conditionally essential, that is,
required in the diet only under certain conditions.
• For example:
-Children and pregnant women have a high rate of protein
synthesis to support growth, require some arginine from the
diet, although it can be synthesized in the body.
-Histidine is essential in the diet of the adult in very
small quantities because adults efficiently recycle histidine.
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CONDITIONALLY ESSENTIAL AMINO ACIDS cont…
-Tyrosine is considered conditionally essential. It is

synthesized from phenylalanine, and it is required in the
diet if phenylalanine intake is inadequate, or if an
individual is congenitally deficient in an enzyme required
to convert phenylalanine to tyrosine (the congenital
disease phenylketonuria).
-Cysteine is also considered as conditionally essential.

It is synthesized by using sulfur from methionine, and it
also may be required in the diet under certain conditions.
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B. ANIMAL SOURCES OF PROTEIN
• These include meat, poultry, milk, and fish
• They have a high quality because they contain all the
essential amino acids in proportions similar to those
required for synthesis of human tissue proteins.
C. PLANT SOURCES OF PROTEIN

• These include wheat, corn, rice, and beans
• They have a lower quality than do animal proteins.
• However, proteins from different plant sources may
be combined in such a way that the result is equivalent
in nutritional value to animal protein.
• For example, wheat which is lysine-deficient but
methionine-rich, may be combined with kidney beans
which is methionine-poor but lysine-rich, to produce an
improved biologic value.
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9. NITROGEN BALANCE
• This occurs when the amount of nitrogen consumed
equals that of the nitrogen excreted in the urine,
sweat, faeces and other excretions.
• Most healthy adults are normally in nitrogen balance.
1. Positive nitrogen balance

• This occurs when nitrogen intake exceeds nitrogen
excretion.
• It is observed during situations in which tissue growth
occurs, for example, in childhood, pregnancy, or during
recovery from an emaciating illness.
• Consumption of excess protein has no physiologic
advantage and so protein consumed in excess of the
body’s needs is deaminated, and the resulting carbon
skeletons are metabolized to provide energy or acetyl
coenzyme A for fatty acid synthesis.
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2. Negative nitrogen balance
• This occurs when nitrogen loss is greater than nitrogen intake.
• It is associated with inadequate intake of dietary protein, lack
of an essential amino acid, or during physiologic stresses, such
as trauma, burns, illness, or surgery.
• If a negative nitrogen balance persists for too long, bodily
function will be impaired by the net loss of critical proteins.
3. NOTE THE PROTEIN-SPARING EFFECT OF
CARBOHYDRATE:
• The dietary protein requirement is influenced by the
carbohydrate content of the diet.
• When the intake of carbohydrates is low, amino acids are
deaminated to provide carbon skeletons for the synthesis of
glucose that is needed as a fuel by the central nervous
system.
• If carbohydrate intake is less than 130 g/day, substantial
amounts of protein are metabolized to provide precursors for
gluconeogenesis. Therefore, carbohydrate is considered to be
“protein-sparing,” because it allows amino acids to be used for
repair and maintenance of tissue protein rather than for
gluconeogenesis. Dr. Mwale, 2023
4. FUEL STORES.
• Any dietary fuel that exceeds the body’s immediate
energy needs is stored, mainly as triacylglycerol
(fat) in adipose tissue, as glycogen (a carbohydrate)
in muscle, liver, and other cells, and, to some extent,
as protein in muscle.
• When we are fasting, between meals and overnight
while we sleep, fuel is drawn from these stores and is
oxidized to provide energy.
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VITAMINS
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VITAMINS
- Vitamins are organic nutrients/molecules that are
required in small quantities in the diet and serve
specialized functions in the body eg normal growth,
maintenance and reproduction..
- They may either not be synthesized at all by the
human body or may be synthesized only at a rate not
consistent with normal health
- They do not undergo degradation to provide energy
- Two forms of vitamins: fat-soluble & water-soluble

- Most of the water soluble vitamins are nearly always
precursors of coenzymes; have more functional groups
that interact with water BUT fat soluble vitamins are
more often encountered as physiological regulators
and antioxidants; have complex hydrocarbon
structures with a few of these having functional
groups 33
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VITAMINS cont….
• Water-soluble vitamins are readily absorbed in the
intestine; excess vitamins are eliminated in urine and
sweat BUT absorption of fat-soluble vitamins
depends on mixed bile salt micelles; excess vitamins
are stored in the liver and thus are not be eliminated
from the body.
• Fat-soluble vitamins deficiencies are most likely in patients

with fat mal-absorption. Fat-soluble vitamins supplements
are most effective when taken with a fatty meal
• Water-soluble vitamins are transported in blood

• Fat-soluble vitamins are transported either as
constituents of lipoproteins or bound to specific
plasma proteins
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ASSIGNMENT 1 TO BE HANDED IN DURING YOUR
TEST/EXAM
Draw a table giving ALL the differences between
water and fat soluble vitamins.
Water-Soluble Vitamins Fat-Soluble Vitamins

1.Are water soluble 1.Are fat soluble
2………… 2………
3…….. 3………
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Fat-soluble Vitamins
1. Vitamin A
• Active forms are retinol, retinal, retinoic acid; found
in abundance in foods of animal origin such as dairy
products, fish and liver.
• Some foods of plant origin contain the antioxidant, β-
carotene (4th form), which the body oxidatively
cleaves in the intestine and is converted to vitamin A
(retinal).
A) Retinal is the aldehyde derived from the oxidation of

retinol (so easily undergo retinal to retinol interconvesion)
• It is pigment that produces a neural signal to the brain
by initiating the response of rod and cone cells of the
retina to light.
• How?
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1. Vitamin A
• Active forms are retinol, retinal, retinoic acid; found
in abundance in foods of animal origin such as dairy
products, fish and liver.
• Some foods of plant origin contain the antioxidant, β-
carotene (4th form), which the body oxidatively
cleaves in the intestine and is converted to vitamin A
(retinal).
A) Retinal is the aldehyde derived from the oxidation of

retinol (so easily undergo retinal to retinol interconvesion)
• It is pigment that produces a neural signal to the brain
by initiating the response of rod and cone cells of the
retina to light.
• How?
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Retinal cont….
• Rhodopsin, the visual pigment of the rod cells in the
retina,has 11-cis retinal specifically bound to the
protein opsin and so when rhodopsin is exposed to light,
a series of photochemical isomerizations occurs,
resulting in the release of all-trans retinal and opsin.
• This process triggers a nerve impulse that is
transmitted by the optic nerve to the brain which
processes the impulse into an image.
• Regeneration of rhodopsin requires isomerization of
all-trans retinal back to 11-cis retinal, which combines
with opsin to form rhodopsin, thus completing the
cycle.
• Similar reactions are responsible for color vision in the
cone cells.
• Note that retinal is also essential for normal
reproduction, ie supporting spermatogenesis in the
male and preventing fetal resorption in the female.
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Vitamin A cont…
B) Retinoic acid is derived from the oxidation of retinol.
• It cannot be reduced in the body, and, therefore,
cannot give rise to either retinal or retinol.
• It binds to nuclear receptors involved in the regulation
of cell growth and differentiation. It also promotes the
development of epithelial tissue including skin. It is
required for the synthesis of the iron transport protein
transferrin.
NOTE:
• Retinoic acid is inactive in maintaining reproduction and
in the visual cycle, although as already mentioned,
promotes growth and differentiation of epithelial cells
• So animals given vitamin A only as retinoic acid from
birth are blind and sterile
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C) Retinol is important for the synthesis of
glycoproteins and mucopolysaccharides. It is found in
animal tissues as a retinyl ester with long-chain fatty
acids
• Also needed for normal growth and for mucus

secretions and so reason why the skin, eyes, and
mucous membranes of the mouth, nose, throat and
lungs depend on vitamin A to remain moist.
• Also required for the synthesis of the transferrin and

for normal reproduction, ie supporting spermatogenesis
in the male and preventing fetal resorption in the
female.
• Vitamin A is also an important antioxidant that may

play a role in the prevention of certain cancers (and
some other aspects of immunity as well). 40
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Absorption and transport of vitamin A to the liver
• Retinyl esters present in the diet are hydrolyzed in the
intestinal mucosa, releasing retinol and free fatty acids.
• Retinol derived from esters and from the cleavage and
reduction of carotenes is re-esterified to long-chain
fatty acids in the intestinal mucosa and secreted as a
component of chylomicrons into the lymphatic system.
• Retinyl esters contained in chylomicron remnants are
taken up by, and stored in, the liver.
Release of Vitamin A from the liver

• When needed, retinol is released from the liver and
transported to extrahepatic tissues by the plasma
retinol-binding protein (RBP).
• The retinol–RBP complex attaches to specific receptors
on the surface of the cells of peripheral tissues,
permitting retinol to enter.
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Mechanism of action of vitamin A
• Retinol is oxidized to retinoic acid which binds with high

affinity to specific receptor proteins present in the
nucleus of target tissues, such as epithelial cells.
• This activated retinoic acid–receptor complex interacts

with nuclear chromatin to regulate gene expression,
resulting in control of the production of specific
proteins that mediate several physiologic functions such
as retinoids controlling the expression of the gene for
keratin in most epithelial tissues of the body.
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Fat-soluble Vitamins cont..
DEFICIENCY OF VITAMIN A
• Night blindness –this involves both time taken to adapt
to darkness and the ability to see in poor light.
• Severe deficency leads to xerophthalmia (progressive
keratinization of the cornea) including pathologic
dryness of the conjunctiva and cornea as Vitamin A is
required for the differentiation and proliferation of the
epithelium of the conjunctiva and cornea.
• When untreated, xerophthalmia results in corneal
ulceration and, ultimately, in blindness because of the
formation of opaque scar tissue.
• Infection usually sets in causing hemorrhaging of the
eyes and permanent loss of vision.
• Very dry, rough skin may indicate a lack of vitamin A,
since retinol and/or retinoic acid down regulate the
synthesis of keratin,
*Excess keratin leads to rough surface in place of moist
and pliable epithelium.
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DEFICIENCY OF VITAMIN A cont…
Keratomalacia
• This is severe eye disorder as a result of vitamin A
deficiency.
• In this condition, normal epithelium is replaced by
inappropriately keratinised epithelium leading to
opaque cornea.
Acne and psoriasis

• Dermatologic problems such as acne and psoriasis can
be effectively treated with retinoic acid or its
derivatives.
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• DEFICIENCY OF VITAMIN A cont….
• Other signs of deficiency include decreased resistance to
infections, due to keratinization of mucosal lining in the
respiratory, gastroinstestinal and genitourinary tracts.
• Fissures which develop in the mucosal lining may allow
microorganisms to enter.
• Vitamin A deficiency can also cause iron deficiency-like anemia,
even in the presence of adequate iron.
VITAMIN A TOXICITY-referred to as Hypervitaminosis A.
• This is due to excess (exceeding 7.5 mg/day) intake of vitamin A
• Symptoms of vitamin A toxicity include dry, itchy skin, bone
pain, nausea, enlarged liver and spleen and loss of appetite.
• Signs of severe overuse over a short period of time include
dizziness, blurred vision and slow growth.
• Other signs may be seen in the nervous system, where a rise in
intracranial pressure may mimic the symptoms of a brain tumor.
• Pregnant women particularly should not ingest excessive
quantities of vitamin A because of its potential for causing
congenital malformations in the developing fetus.
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Xerophthalmia
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Fat-soluble Vitamins cont……….
2. Vitamin D
•Involved in homeostasis
•Found in animals (as cholecalciferol-vitamin D3), oily fish(e.g.,
herring, salmon,sardines), in cod liver oil, milk and other diary
products
•Found in plants (ie Ergocalciferol-vitamin D2),
Effect in man:
i)increases Ca absorption from intestines How? Calcitriol
(formed from the hydroxlation of Cholecalciferol) binds to a
cytosolic receptor. The calcitriol–receptor complex then
moves to the nucleus where it selectively interacts with the
cellular DNA resulting into enhanced calcium uptake by an
increased synthesis of a specific calcium-binding protein.
ii)causes Ca mobilization from bone
iii)reduces excretion of calcium (by stimulating
resorption in the distal renal)
iv) increases phosphate excretion by kidneys
•Note that sunlight converts 7-dehydrocholesterol to a form
of vitamin D, cholecalciferol, in the skin and therefore not
strictly a vitamin but an endogenous vitamin precursor.
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2. Vitamin D cont……
v)Vitamin D is also involved in insulin secretion, synthesis
and secretion of parathyroid and thyroid hormones (via
interaction with intracellular receptor proteins),
modulation of cell proliferation (ie interacts with DNA in
the nucleus of target cells in a manner similar to that of
vitamin A, and either selectively stimulates gene
expression or specifically represses gene transcription),
normal functioning of immune system, regulation of blood
pressure and normal neuromuscular function.
DEFICIENCY OF VITAMIN D
•In growing children, deficiency include rickets (soft and
pliable bones) due to improper mineralization of osteoid
matrix and cartilage.
•Symptoms of rickets include delayed growth, pain in the
spine, pelvis and legs and muscle weakness.
•Because rickets softens the growth plates at the ends
of a child’s bones, it can cause skeletal deformities such
as bowed legs, abnormally curved spine, thickened wrists
and ankles and breast bone projection. 48
Dr. Mwale, 2023
DEFICIENCY OF VITAMIN D cont…..
•Vitamin D deficiency in adults may result in osteomalacia
caused by demineralization of preexisting bones causing it to
become softer and increased susceptibility to fracture.
• Insufficient exposure to daylight and/or deficiencies in
vitamin D consumption occur predominantly in infants and
the elderly.
• Renal osteodystrophy is chronic renal failure that results
from the decreased ability to form the active form of
vitamin D
• Supplementation with calcitriol is an effective therapy for
these conditions.
VITAMIN D TOXICITY
•Signs of vitamin D toxicity(excess of 100,000 IU for weeks
or months) include excess calcium in the blood, contraction of
blood vessels, high blood pressure, and calcinosis
(calcification of soft tissues).
•Toxicity can also lead to hypercalcemia leading to
hypercalciuria resulting in formation of renal stones
49
Dr. Mwale, 2023
RICKETS
Osteomalacia
50
Dr. Mwale, 2023
Fat-soluble Vitamins cont…….
2. Vitamin E
•3 forms exist:α-, β-, γ- tocopherol/tocotrienols; most
active is the α-form
• Vitamin E benefits the body by acting as an antioxidant,
in cell membranes and plasma lipoproteins.
• It reacts with the lipid peroxide radicals formed by
peroxidation of polyunsaturated fatty acids.
• They also prevent the oxidation of LDL, which maybe
important in reducing the risk of cardiovascular diseases
since the oxidized form of LDL is atherogenic.
•Source of vitamin E in the diet comes from vegetable oil
(soybean, corn, cottonseed, and safflower), fruits and
vegetables, grains, nuts (almonds and hazelnuts), seeds
(sunflower) and fortified cereals.
•In rats, it is known to act as an anti-sterility vitamin.
51
Dr. Mwale, 2023
Fat-soluble Vitamins cont…….
2. Vitamin E cont….
VITAMIN E DEFICIENCY
• Though rare it can only occur in premature infants and
in those unable to absorb or transport fats. It causes
fragile red blood cells and the appearance of abnormal
cellular membranes.
VITAMIN E TOXICITY
• Vitamin E can act as an anticoagulant and may increase
the risk of bleeding problems.
• Occasionally, muscle weakness, fatigue, nausea, and

diarrhea have been reported to occur.
• However, Vitamin E is the least toxic of the fat-soluble

vitamins even at doses of 300 mg/day
52
Dr. Mwale, 2023
2. Vitamin K
•2 forms exist:K1 & K2 derived from menadione
• Vitamin K2 is naturally produced by intestinal bacteria, and
plays an essential role in normal blood clotting (prothrombin
biosynthesis)
•It is also required for the carboxylation of glutamate residues
(formation of γ-carboxyglutamate,Gla) in calcium-binding bone
proteins which is required for normal bone mineralization.
How?
•As stated, Vit. K–dependent carboxylation of glutamic acid
residues is central to synthesis of prothrombin and blood
clotting factors II, VII, IX, and X inactive precursor molecules
which forms a mature clotting factor that is capable of
subsequent activation.
•Note that the carboxylation is sensitive to inhibition by

dicumarol, an anticoagulant and by warfarin, a synthetic analog
of vitamin K.
•There is also an interaction of prothrombin with platelets,

where the Gla residues of prothrombin bind to calcium ions
(prothrombin–calcium complex) which is then able to bind to
phospholipids that are essential for blood clotting on the surface
of platelets.
53
Dr. Mwale, 2023
Vitamin K cont…
•Sources of vitamin K are green, leafy-vegetables
(spinach, cauliflower, cabbage and broccoli), and certain
vegetables oils (including soybean oil, canola oil and olive
oil).
•Animal foods, in general, contain limited amounts of

vitamin K.
•Vit.K1 available in green leaves of plants as phylloquinone
54
Dr. Mwale, 2023
VITAMIN K DEFICIENCY
• Most easily detectable symptom of vitamin K
deficiency is
increased coagulation time.
• Deficiency is most commonly seen in newborn babies
lacking the intestinal bacteria to produce vitamin K.
Because human milk provides only about one fifth of the
daily requirement for vitamin K, it is recommended that
all newborns receive a single intramuscular dose of
vitamin K as prophylaxis against hemorrhagic disease.
• People taking antibiotics may lack vitamin K temporarily
(hypoprothrombinemia) because intestinal bacteria are
sometimes killed as a result of long-term use of
antibiotics
VITAMIN K TOXICITY
• Excessive amounts can cause the breakdown of red
blood cells (hemolytic anemia and jaundice in infants.)
and liver damage
• Prolonged administration of large doses of synthetic
vitamin K (menadione) is no longer used to treat vitamin
K deficiency therefore
soprotection.com 55
Dr. Mwale, 2023
SCHOOL OF MEDICINE
WATER SOLUBLE VITAMINS

Conference Room, Adult Extension Studies Building,
Ridgeway Campus
1
OBJECTIVE of the topic:
• Describe normal nutrition and deviations from the normal state
which can lead to diseases.
• NUTRITION
– Definition
– Importance of Nutrition
• Importance of food related to death
• Factors that contribute to death
– Classification of Nutrients
– Why we eat what we eat?
– Dietary Reference Intakes
– Dietary carbohydrate
– Dietary fats
– Dietary proteins
– Nitrogen balance
– Fuel stores
• VITAMINS
– Overview
– Fat-soluble vitamins
• A,D,E,K
– Water-soluble vitamins
• Thiamine (Vitamin B1), Riboflavin (Vitamin B2), Pantothenic acid (Vitamin B5),
Niacin (Nicotinic acid B3), Pyridoxine (Vitamin B6), Biotin (Vitamin B7),
Cobalamin (Vitamin B12), Folic acid (pteroylglutamic acid B9)
• Vitamin C Dr. Mwale, 2023
Water-Soluble Vitamins
***Includes vitamins B and C***
• They function as coenzymes that help the body obtain
energy from food.
• Because of their water solubility these vitamins can
excreted in urine and sweat and rarely accumulate in
toxic concentrations.
The vitamins of the B complex

• Thiamine (Vitamin B1), Riboflavin (Vitamin B2),
Pantothenic acid (Vitamin B5), Niacin (Nicotinic acid
B3), Pyridoxine (Vitamin B6), Biotin (Vitamin B7),
Cobalamin (Vitamin B12), Folic acid (pteroylglutamic
acid B9)
• The B vitamins are also important for normal appetite,

good vision, and healthy skin, nervous system, and red
blood cell formation. 3
Dr. Mwale, 2023
Vitamin B1 (Thiamine)
Source-all plant and animal tissues used as food especially
unrefined cereal grains and meat including peas,
pork, liver, legumes and enriched products like bread,
pasta and rice
• Has a key role in carbohydrate metabolism ie participation
in the oxidative decarboxylation of pyruvate and α-
ketoglutarate, a key role in energy metabolism of most
cells, especially in tissues of the nervous system
• It is also important in transmission of nerve impulses by

acting as a phosphate donor for phosphorylation of the
nerve membrane sodium transport channel.
• It also helps to release energy from foods as Thiamine

pyrophosphate,( which is the biological active form of the
vitamin formed by the transfer of a pyrophosphate group
from ATP to thiamine) by serving as a coenzyme in
enzymatic reactions transferring an activated aldehyde unit
4
Dr. Mwale, 2023
Vitamin B1 (Thiamine) cont….
THIAMIN DEFICIENCY
-Deficiency results into a decreased production of ATP and
consequently, impaired cellular function.
-Chronic peripheral neuritis
-Beriberi (severe muscle weakness and muscle wasting,

delirium, opthalmoplegia and memory loss) .When this is
coupled with edema, the disease is called wet beriberi. The
deficiency syndrome can have a rapid onset in nursing
infants whose mothers are deficient in thiamine
-Wernicke’s –Korsakoff encephalopathy with psychosis –
especially associated with alcohol (because excess alcohol
consumption often replaces food or meals). Symptoms
include ataxia (unsteady gait while walking and general
inability to achieve control of motor functions), mental
confusion, ophthalmoplegia (loss of eye coordination).
• Major symptoms of mild thiamine deficiency include: loss of

appetite, constipation, nausea, irritability and fatigue.
• Treatment can be by thiamine supplementation. 5
Dr. Mwale, 2023
Vitamin B2 (Riboflavin)
• Synthesized by all plants and microorganisms (not by
higher animals)
• Has a central role in energy yielding metabolism; it

also promotes good vision, and healthy skin
• Has a central role in mobilization of iron and also helps
to convert the amino acid tryptophan into niacin.
• Flavin coenzymes (FAD and FMN) are the two most

active forms of the vitamin
• FAD and FMN are electron carriers in oxidoreduction
reactions formed by the transfer of an adenosine
monophosphate moiety from ATP to FMN
• Found in key enzymes in fatty acid and amino acid
oxidation, including the citric acid cycle
• The main dietary precursors of riboflavin are milk and

dairy products, liver, eggs, dark green vegetables,
legumes, whole and enriched grain products,
– Because of its intense colour riboflavin is sometimes
used as a food additive
6
Dr. Mwale, 2023
Vitamin B2 (Riboflavin) cont………….
• Riboflavin deficiency usually occurs along with other

vitamins deficiences and is most common in alcoholics,
malignancy, hypothyroidism, and in the elderly.
– Symptoms include cracks at the corners of the

mouth, dermatitis on nose and lips, light sensitivity,
cataracts, and a sore throat, glossitis, angular
stomatitis, and seborrheic dermatitis
– Riboflavin deficiency can occur in infants receiving

phototherapy because riboflavin is destroyed by
light in the skin
7
Dr. Mwale, 2023
Niacin(Nicotinic acid B3)
• It can be synthesized from the essential amino acid
tryptophan-pathway for synthesis requires pyridoxal
phosphate, coenzyme form of pyridoxine.
• Is part of NAD+ and NADP+ coenzymes in oxidation-reduction
reactions(energy production) ie active forms of the vitamin.
• The reduced forms of NAD+ and NADP+ are NADH and
NADPH, respectively.
• Also involved in normal enzyme function, digestion, promoting
normal appetite,healthy skin, and nerves
• The major sources of niacin are tryptophan-containing
proteins and those foods containing nicotinic acid per se
(unrefined cereals, vegetables, milk, liver, fish, poultry, meat,
peanuts,).
• Corn is very poor in tryptophan and available niacin and so
diets in which corn is a major source of protein can result in a
deficiency called pellagra, which affect the skin,
gastrointestinal tract, and CNS.
• The symptoms of pellagra progress through the three Ds:
photosensitive dermatitis, evidence of dementia, diarrhea and,
if untreated, death.
• Deficiency is associated with with alcoholism, protein
malnourishment, low calorie diets, and diets high in refined
carbohydrates. 8
Dr. Mwale, 2023
Niacin(Nicotinic acid B3)
• Deficiency cont…
• Niacin strongly inhibits lipolysis in adipose tissue

resulting in hyperlipidemia
• Niacin also useful in treating hyperlipoproteinemia
• The liver normally uses these circulating fatty acids as

a major precursor for triacylglycerol synthesis BUT
the activity of niacin causes a decrease in liver
triacylglycerol synthesis, which is required for VLDL
production. LDL is derived from VLDL in the plasma.
• So both VLDL and LDL are elevated.

9
Dr. Mwale, 2023
Vitamin B6 (Pyridoxine)
• Vitamin B6 is a generic name for the dietary precursors
of the active coenzyme form pyridoxal phosphate (PLP)
• Is important in amino acid and glycogen metabolism, and
in steroid hormone action
• 80% of total body vitamin B6 is present as pyridoxal
phosphate (or pyridoxal)in muscle, associated mostly with
glycogen phosphorylase
• The 3rd form is pyridoxamine.
• Pyridoxine, pyridoxal, and pyridoxamine, are all
derivatives of pyridine which differ only in the nature of
the functional group attached to the ring.
• Pyridoxine occurs primarily in plants, whereas pyridoxal
and pyridoxamine are found in foods obtained from
animals and so sources include pork, meats, whole grains
and cereals, legumes, and green, leafy vegetables.
• Roles-Coenzyme for many enzymes in amino acid

metabolism and coenzyme for glycogen phosphorylase in
muscle.
• It is also required for the synthesis of
neurotransmitters serotonin, norepinephrine,
epinephrine, and γ-aminobutyrate (GABA), and also
synthesis of sphingomyelin. 10
Dr. Mwale, 2023
Vitamin B6 (Pyridoxine)
• Deficiency has been observed in new-born infants fed
formulas low in B6, in women taking oral contraceptives.
• Also common in alcoholics resulting in side-roblastic
anaemia, peripheral neuropathy and seizures.
• Other classic clinical syndrome for vitamin B6
deficiency is a dermatitis-like eruption, glossitis with
ulceration, angular cheilitis, conjunctivitis, intertrigo,
and neurologic symptoms like, confusion, and
neuropathy (due to impaired sphingosine synthesis) and
side roblastic anemia (due to impaired heme synthesis).
– Isoniazin and penicillamine can precipitate vitamin
B6 deficiency
Toxicity of pyridoxine
• Pyridoxine is the only water-soluble vitamin with
significant toxicity.
• Neurologic symptoms (sensory neuropathy) occur at
intakes above 200 mg/day, an amount more than 100
times the RDA
• Substantial improvement, but not complete recovery,
occurs when the vitamin is discontinued. 11
Dr. Mwale, 2023
Pantothenic acid(Vitamin B5)
• Pantothenic acid functions as a constituent of coenzyme A
(CoA) which takes part in citric acid cycle,fatty acid
synthesis and oxidation, acetylations, cholesterol synthesis.
• CoA contains a thiol group that carries acyl compounds.
• Examples of such structures are succinyl CoA, fatty acyl
CoA, and acetyl CoA.
• Pantothenic acid is also a component of the acyl carrier
protein (ACP) domain of fatty acid synthase.
• Also aids in the formation of hormones and cholesterol,
and the metabolism of fats and carbohydrates from food.
• It is also critical in the production of RBCs, and
maintaining healthy GIT.
• Sources include liver, kidney, meats, egg yolk, whole grains,
and legumes.
Deficiency
– is uncommon due to its wide availability in most foods.
– But symptoms may include fatigue, depression,
insomnia,vomiting, stomach pains.
12
Dr. Mwale, 2023
Biotin(Vitamin B7)
• Biotin helps release energy from carbohydrates and
aids in the metabolism of fats, proteins and
carbohydrates from food.
• It is also a coenzyme for enzymes participating in
different metabolic reactions-particularly useful in
carboxylation reactions.
• Sources of Biotin include liver, kidney, egg yolk, milk,
most fresh vegetables, yeast breads and cereals.
Biotin is also made by intestinal bacteria.
• Deficiency is uncommon under normal circumstances

because the vitamin is widely distributed in food, but
symptoms include fatigue, loss of appetite, nausea,
vomiting, depression, muscle pains, heart abnormalities
and anemia.
• Can be found in people on paranteral nutrition for a

long time
• *Eating abnormally large amounts of uncooked egg

white which contains avidin, a glycoprotein that binds
to biotin and renders it unavailable for absorption*13
Dr. Mwale, 2023
Folic acid (pteroylglutamic acid B9)
• It aids in rapid cell division and growth, promoting RBC
formation, and lowering the risk for neural tube birth
defects.
• The active form of folic acid is tetrahydrofolate,

which is a one carbon carrier from donors such as
serine, glycine, and histidine and transfers them to
intermediates in the synthesis of amino acids, purines,
and thymidine monophosphate (TMP, a pyrimidine
found in DNA).
• In its active form it is therefore a coenzyme involved

in various reactions such as in metabolism of amino
acids, purines and pyrimidines.
• It is synthesized by intestinal bacteria but other

sources include liver, kidney, dark green leafy
vegetables, meats, fish, whole grains, fortified grains
and cereals, legumes, and citrus fruits.
14
Dr. Mwale, 2023
Folic acid (pteroylglutamic acid B9) cont…
• Deficiency inhibits DNA synthesis by decreasing availability of
purines and thymidine, affecting cell growth as cells can not
divide-symptoms also include megaloblastic anemia (abnormally
large macrocystic RBC with fragile membranes).
• Folate deficiency leads to arrest of cells in S phase, resulting
in abnormal size and shape RBC and a slower maturation .
• In pregnant or women of child bearing age deficiency may
result in the delivery of a baby with neural tube defects
either due to defects in cell division or gene regulation during
development (Spina bifida and anencephaly)
• Folic acid supplementation before conception and during the
first trimester has been shown to significantly reduce the
defects.
• Therefore, all women of childbearing age are advised to
consume 0.4 mg/ day of folic acid.
• Adequate folate nutrition must occur at the time of
conception because critical folate-dependent development
occurs in the first weeks of fetal life - at a time when many
women are not yet aware of their pregnancy.
• However, there is an association of high-dose supplementation
with folic acid (>0.8 mg/day) and an increased risk of cancer.
Thus, supplementation is not recommended for most middle-
aged or older adults.
• Alcoholism and mal-absorption can also cause folate
deficiency
15
Dr. Mwale, 2023
Folic acid (pteroylglutamic acid B9) cont…
Deficiency cont..
• Folic acid deficiency is the second major category of
nutritional anemia also refered to as macrocytic
• These macrocytic anemias are commonly called
megaloblastic because their deficiency causes
accumulation of large, immature red cell precursors,
known as megaloblasts, in the bone marrow and the
blood.
Note that :
1. Vitamin B12 deficiency is as megaloblastic
2. Nutritional anaemias are caused by inadequate intake
of one or more essential nutrients—can be classified
according to the size of the red blood cells or mean
corpuscular volume observed in the individual.
3. The second type of Nutritional anaemias is therefore
the Microcytic anemia, caused by lack of iron, is the
most common form of nutritional anemia.
16
Dr. Mwale, 2023
Vitamin B12 (Cobalamin)
• Found only in foods of animal origin (meats, liver, kidney,
fish, eggs, milk and milk products, oysters, shellfish)
• Plants do not contain Vitamin B12
• It is the most complex of all the vitamins.
• Synthesized exclusively by bacteria but is present in
normal animal liver-a small amount is synthesized by colon
bacteria.
• Is involved in isomerization of methylmalonyl CoA to
succinyl CoA, a Krebs Cycle intermediate, and
remethylation of homocystein to methionine. Also aids in
the building of genetic material (synthesis of DNA),
metabolism of fatty acids and amino acids, production of
normal red blood cells, and maintenance of the nervous
system.
• Absorption of Vit B12 requires intrinsic factor, a
glycoprotein secreted by parietal cells of the gastric
mucosa. It binds tightly to intrinsic factor and in this
form its absorbed in the ileum
– In the blood it binds to transcobalamin II and other
plasma proteins.Transcobalamin II directs the vitamin
to the tissues where it is required and prevents its
renal secretion
17
Dr. Mwale, 2023
Vitamin B12 (Cobalamin) cont………
• Deficiency most commonly affects strict vegetarians,
infants of vegan mothers, and the elderly unless the
food is contaminated by bacteria or fecal matter
• Pernicious anemia is an autoimmune disease that

destroys parietal cells in the stomach and so Vitamin
B12 cannot be absorbed through their stomach lining.
This deprives the patient of intrinsic factor and
neither dietary nor biliary Vit B12 can be absorbed.
This can be treated through vitamin B12 injections.
• Pernicious anemia shows 2 types of abnormalities:

-Megaloblastic anemia , similar to that of folate
deficiency
-Neurological dysfunction caused by demyelination of
peripheral nerves and the spinal cord (irreversible)
• Vitamin B12 deficiency blocks the metabolism of folic

acid, leading to functional folate deficiency. This
impairs erythropoiesis, causing immature precursors of
erythrocytes to be released into the circulation
(megaloblastic anemia). 18
Dr. Mwale, 2023
Vitamin B12 (Cobalamin) cont………
• However, many (though not all) of the effects of functions of B12
can be replaced by sufficient quantities of folic acid (vitamin
B9), since B12 is used to regenerate folate in the body.
• Additionally, the recommended daily allowance for Vit B12 is 2.4
μg; btn 1 and 10 mg of Vit B12 is stored in the body, most of this
in the liver. Therefore a sudden switch to a vitamin B12-free diet
will cause serious deficiency only after a few decades including in
individuals who have had a partial or total gastrectomy and can no
longer absorb the vitamin.
• Impaired intestinal absorption causes deficiency within 2-6 years
• Note that every day between 1 -10 μg of Vit B12 is secreted into
the bile and most of it is reabsorbed with the help of intrinsic
factor
• Most cases of Vit B12 deficiency are caused by mal-absorption
NOTE
• When the vitamin is deficient, unusual fatty acids accumulate and
become incorporated into cell membranes, including those of the
nervous system. This may account for some of the neurologic
manifestations of vitamin B12 deficiency.
• Pernicious anemia can be treated by giving high-dose B12 orally, or
intramuscular (IM) injection of cyanocobalamin.
19
Dr. Mwale, 2023
Vitamin C
• Ascorbic acid or ascorbate, is essential for proper
functioning of the body
• It benefits the body by holding cells together through
collagen synthesis, aids in wound healing, bone and
tooth formation, strengthening blood vessel walls.
• It also improves immune system function, and
increasing absorption and utilization of iron by
reducing it to ferrous state in the stomach.
• It enhances the utilization of folate by converting it
to tetrahydrofolate.
• It is required for the synthesis of norepinephrine.
• It works with vitamin E as an antioxidant, and plays a
crucial role in neutralizing free radicals throughout
the body
• It is a reducing agent and a scavenger of free radicals;
readily destroyed by heat and light
• Available in fresh fruits (citrus fruits, such as orange,
kiwi fruit, grapefruit, sweet red pepper and vegetables
20
Dr. Mwale, 2023
Vitamin C cont….
It is also the coenzyme for two hydroxylases

• Dopamine β-hydroxylase, a copper containing
enzyme involved in synthesis of catacholamines
• Those involved in modification of precursor
proteins
eg Proline and lysine hydroxylases
It is also required for modifications of procollagen

to collagen
• Deficiency leads to scurvy – skin changes, fragility

of blood vessels, gum decay, bone fracture; due to
deficiency of collagen ie it causes a loss of
collagen strength throughout the body.
-Loss of collagen results in loose teeth, bleeding
and swollen gums, and improper wound healing. 21
Dr. Mwale, 2023
ASSIGNMENT 2 TO BE HANDED IN DURING YOUR
TEST/EXAM
Draw a table summarising the vitamins.
Vitamin Function Deficiency Disease

A(Retinol) Visual pigment in the Night blindness,
retina,… xerophthalmia,…
22
Dr. Mwale, 2023
Fortified Foods.
• Fortified foods are those that have nutrients added

to them thereby boosting vitamin and mineral intake.
• Introduced in the 1930s
Examples
• Milk fortified with Vit D and A
• Fruit juice fortified with Calcium
• Vegen foods fortified with Vit B12
• Bread fortified with folic acid
• Eggs fortified with omega-3 fatty acids
• Some Soy products fortified with calcium, Vit. A and D
• Unflavored yogurt
• Enriched foods are means nutrients that were lost

during processing are added back in
Examples
Many refined grains such as wheat flour have folic acid,
riboflavin and iron-added back after processing 23
Dr. Mwale, 2023
THANK YOU
24
Dr. Mwale, 2023
soprotection.com
GLYCOLYSIS
SHARI R. BABU
shari.babu@unza.zm
DEPT. OF PHYSIOLOGICAL SCIENCES
SCHOOL OF MEDICINE, UNZA
INTRODUCTION
• Glucose is the predominant metabolic fuel in almost all cells.
• It is especially important for;
• cells that depend on anaerobic metabolism,
• cells that lack mitochondria, and
• tissues such as brain that under normal conditions cannot use other
metabolic fuels.
• The concentration of glucose in the plasma is controlled by number of

hormones, mainly by two peptide hormones: insulin and glucagon
OVERVIEW OF GLYCOLYSIS
• Glycolysis is the major pathway of carbohydrate metabolism that occurs in

almost all organisms from plants, microbes to mammals.
• Also called Embden-Meyerhof-Parnas (EMP) pathway.
• Glycolysis can occur under aerobic as well as anaerobic conditions.
• Glycolysis is the first step in cellular respiration and occurs in the cytosol.
• This pathway oxidizes glucose to provide energy (in the form of ATP) and
intermediates for other metabolic pathways.
• Glucose (6-carbon molecule) is split into two molecules of pyruvate (3-carbon

compound).
• Pyruvate is the end product of aerobic glycolysis in cells with mitochondria

and an adequate supply of oxygen.
Cellular Respiration
• In cells lacking mitochondria or under anaerobic conditions, pyruvate is reduced
to lactate.
• This conversion of glucose to lactate is called anaerobic glycolysis because it

can occur without the participation of oxygen.
• Anaerobic glycolysis allows the production of ATP in tissues that lack

mitochondria (for example, red blood cells) or in cells deprived of sufficient
oxygen.
TRANSPORT OF GLUCOSE
Glucose cannot diffuse directly into cells but enters by one of two transport
mechanisms: a Na+-independent, facilitated diffusion transport system or a Na+-
monosaccharide co-transporter system.
Sodium-dependent glucose co-
• It is a specific transporter located on the luminal transporter (SGLT)
side of intestinal cells and proximal tubule in
kidneys.
• SGLT uses the sodium concentration gradient

maintained by Na+/K+ ATPase.
• Sodium moves down its concentration gradient

into the cell, taking glucose with it each time.
• SGLT1: One molecule of glucose is absorbed

together with two molecules of sodium in the
small intestine and kidney.
• SGLT1 is responsible for reabsorbing nearly

3% of the filtered glucose in PCT.
• SGLT2: One molecule of glucose is absorbed

together with one molecule of sodium in PCT.
• SGLT2 is responsible for 97% of glucose

reabsorption in the proximal tubule.
+
Na -independent facilitated diffusion transport
• This system is mediated by a family of glucose transporters present in cell membranes

(GLUT).
GLUT-1 is abundant in GLUT-2 is found in the liver, GLUT-3 is found in most human
most human cells, RBCs, kidney, pancreatic b cells and cells, CNS, and placenta.
CNS, cornea, blood brain small intestine. • High affinity for glucose
barrier, placenta, and fetal • Transports all monosaccharides.
tissue. • Glucose sensor.
• High affinity for glucose • Low affinity for glucose (glucose
only diffuses at high
concentrations).
• Bidirectional transporter: allows
hepatocytes to uptake glucose
for glycolysis and release
glucose during gluconeogenesis
and glycogenolysis.
GLUT 1, 2 and 3 are insulin-independent glucose transporters.

• GLUT-4 is abundant in adipocytes, heart and skeletal muscles.
• Insulin stimulates movement of GLUT4 (stored in vesicles) to plasma

membranes for glucose uptake.
• The number of active GLUT-4 transporters is increased by insulin.
• Physical exercise also induces the translocation of GLUT4 into the

plasma membrane of skeletal muscle, in an insulin-independent
manner.
Glycolysis Has Two Phases
• The breakdown of the six-carbon glucose into two molecules of the three-
carbon pyruvate occurs in ten steps.
• The first five reactions constitute the preparatory (investment) phase, which
is the energy requiring phase.
• In this phase ATP is invested, and the carbon chains of all the hexoses
intermediates are converted into a common product, glyceraldehyde 3-
phosphate.
• The last five reactions form the energy–releasing phase, called the payoff
phase.
• Pyruvate is formed at the end of this phase along with ATP per molecule.
• Energy is also conserved in this phase in the formation of NADH molecule.

SB13
1. Phosphorylation of Glucose: Phosphorylated sugar molecules do not readily

cross cell membranes, because there are no specific transporters for these
compounds.
• They remain effectively trapped in the cells.
• Phosphorylation of glucose ensure the intracellular concentration of
glucose is low allowing for the continued uptake of glucose by the cells.
• Glucose is activated by its phosphorylation at C-6 to yield glucose 6-

phosphate, with ATP as the phosphoryl donor.
S1
Slide 12
SB13 Glucokinase regulatory protein

(GKRP) in the liver regulates the activity of glucokinase through reversible binding. In the presence of
fructose 6- phosphate, glucokinase is translocated into the nucleus and binds tightly to the regulatory
protein, thus rendering the enzyme inactive. When glucose levels in the blood (and also in the
hepatocyte, as a result of GLUT-2) increase, glucokinase is released from the regulatory protein, and the
enzyme re-enters the cytosol where it phosphorylates glucose to glucose 6-phosphate. GKRP also acts as
a competitive inhibitor, competeing with glucose to bind to the active site of glucokinase.
Shari Babu, 15/03/2021
S1 glucose 6 phosphate contains a energy rich bond

Shari, 23/06/2023
• In liver parenchymal cells and β-cells of the pancreas, glucokinase is the
predominant enzyme responsible for phosphorylation of glucose.
• The function of glucokinase in the liver is to provide glucose 6-phosphate in

excess of requirements for glycogen synthesis.
• Glucokinase of the pancreatic β-cells act as glucose sensor and regulate the
secretion of insulin.
SB15
Slide 14
SB15 Glucose-6-phosphate is an important compound that can be diverted to many metabolic pathway
(glycolysis, gluconeogensis, pentose phosphate pathway, and glycogenesis).
2. Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate: The
enzyme phosphohexose isomerase (phosphoglucose isomerase) catalyzes
the reversible isomerization of glucose 6-phosphate to fructose 6-
phosphate.
3. Phosphorylation of Fructose 6-Phosphate to Fructose 1,6-
Bisphosphate: Phosphofructokinase-1 (PFK-1) catalyzes the transfer of a
phosphoryl group from ATP to fructose 6-phosphate to yield fructose 1,6-
bisphosphate:
• The PFK-1 reaction is essentially irreversible under cellular conditions, and it is

the first “committed” step.
4. Cleavage of Fructose 1,6-Bisphosphate: The enzyme aldolase catalyzes a
reversible reaction. Fructose 1,6-bisphosphate is cleaved to yield two different
triose phosphates, glyceraldehyde 3-phosphate, an aldose, and
dihydroxyacetone phosphate, a ketose:
5. Isomerization of the Triose Phosphates: Only glyceraldehyde 3-phosphate,
can be directly degraded in the subsequent steps of glycolysis. The other
product, dihydroxyacetone phosphate, is rapidly and reversibly converted to
glyceraldehyde 3-phosphate by the fifth enzyme of the sequence, triose
phosphate isomerase.
• This isomerization results in the net production of two molecules of

glyceraldehyde 3-phosphate from the cleavage products of fructose 1,6-
bisphosphate.
6. Oxidation of Glyceraldehyde 3-Phosphate to 1,3-Bisphosphoglycerate: The
first step in the payoff phase is the oxidation and of glyceraldehyde 3-phosphate
along with the attachment of a phosphate group to form 1,3-
bisphosphoglycerate, catalyzed by glyceraldehyde 3- phosphate
dehydrogenase:
• This is the first oxidation-reduction reaction of glycolysis.

• Hyperglycemia inhibits glyceraldehyde 3-phosphate dehydrogenase.
• Hyperglycemia promotes overproduction of reactive oxygen species

(superoxide) which activates poly(ADP-ribose) polymerase.
• This enzyme adds multiple ADP-ribose molecules to the free sulfhydryl

groups of active site cysteine residues of glyceraldehyde 3-phosphate
dehydrogenase.
• Causing inhibition of glyceraldehyde 3-phosphate dehydrogenase.

7. Synthesis of 3-Phosphoglycerate with the formation of ATP: When 1,3-
BPG is converted to 3-phosphoglycerate, the high-energy phosphate group of
1,3-BPG is used to synthesize ATP from ADP. This reaction is catalyzed by
phosphoglycerate kinase, which is reversible.
• This is an example of substrate-level phosphorylation (SLP).

• SLP is a mechanism of ATP formation which involves the transfer of a
phosphate (Pi) from an energy rich metabolic intermediate to ADP to form
ATP.
• For SLP to occur, the donor molecule must be a high energy phosphate
containing compound; the energy released during the hydrolytic release of the
Pi must be high enough to attach the Pi to ADP.
S
P
ENZYME ENZYME
ATP
ADP
• Mechanism of arsenic poisoning: Arsenate prevents ATP production
without inhibiting the pathway itself.
• Due to its structural similarity to phosphate, it interacts with the Cys-SH group
of glyceraldehyde 3-phosphate dehydrogenase, forming a complex (1-arseno-
3-phosphoglycerate) that spontaneously hydrolyzes to form 3-
phosphoglycerate.
• By bypassing the synthesis of 1,3- BPG, ATP is not formed by

phophoglycerate kinase.
• Thus, there is no net ATP synthesis occurring when glycolysis takes place
in the presence of arsenate
8. Conversion of 3-Phosphoglycerate to 2-Phosphoglycerate: The enzyme
phosphoglycerate mutase catalyzes a reversible shift of the phosphoryl
group between C-2 and C-3 of glycerate; Mg2+ is essential for this reaction:
9. Dehydration of 2-Phosphoglycerate to Phosphoenolpyruvate: In this
reaction, enolase promotes reversible removal of a molecule of water from 2-
phosphoglycerate to yield phosphoenolpyruvate (PEP):
• PEP is an unstable molecule and can transfer its phosphate group more
easily than 2-phosphoglycerate.
• Fluoride is a potent inhibitor of enolase.
10. Transfer of the Phosphoryl Group from Phosphoenolpyruvate to ADP:
The last step is the transfer of the phosphoryl group from
phosphoenolpyruvate to ADP, catalyzed by pyruvate kinase, which requires
K and either Mg2+ or Mn2+ . This reaction is irreversible.
SUMMARY
• Glycolysis takes place in the cytosol.
• C6 molecule is split into two C3 molecules.
• There are TEN reactions. Three are irreversible and seven reversible
reactions.
• Overall reaction: Glucose + 2ATP + 2NAD+ = 2 Pyruvate + 4ATP + 2NADH
• Key enzymes: Hexokinase (Glucokinase), Phosphofructokinase-1, and

Pyruvate kinase.
REGULATION OF GLYCOLYSIS
• Hexokinase (glucokinase), PFK-1 and pyruvate kinase can regulate the rate
of glycolysis according to the physiological need of the body.
• Regulation can be hormonal (via cAMP), or it can be allosterically (via

reactant or product).
S3
S4
• Glucagon can inhibit the synthesis of hexokinase, and hence inhibits the first
step of glycolysis.
• This results in increased blood glucose levels.
• Insulin increases activity of GLUTs and increases synthesis of hexokinase.
• This results in decreased blood glucose levels.
• Hexokinase is inhibited by its product, glucose-6-phosphate and activated by

AMP and ADP.
• Glucokinase is inhibited by fructose-6-phosphate.

Slide 29
S3 Hexokinase has high affinity for glucose. This ensures that even at low glucose concentration glycolysis
occurs in extrahepatic tissues.
Shari, 27/06/2023
S4 Glucokinase has low affinity for glucose. This means that it will bind to glucose only when the glucose
concentration is very high.
Shari, 27/06/2023
• PFK-1 is inhibited allosterically by:
S5
• Elevated levels of ATP and citrate.
• PFK-1 is also inhibited high proton concentration. This make sures there is
no cell damaging effect of decreased pH as a result of anaerobic pyruvate
metabolism.
• PFK-1 is activated allosterically by high concentrations of AMP and fructose

2,6-bisphosphate.
S6
• Fructose 2,6-bisphosphate is the most potent activator of PFK-1 and can

activate the enzyme even when ATP levels are high.
S7
• Pyruvate kinase is inhibited by ATP and alanine.
• It is activated by fructose 1,6-bisphosphate.

Slide 30
S5 Citrate is formed in the CAC from acetyl CoA which is produced from pyruvate. Its high-level signals that
the substrate requirements for CAC has been met.
Shari, 27/06/2023
S6 F2,6BP is not a glycolytic intermediate. It is not interconvertible with F1,6BP.

Shari, 27/06/2023
S7 Alanine can be produced from pyruvate. Its formation signals that the substrate requirements for other
metabolic pathways have been met
Shari, 27/06/2023
SB10
In clinical setting there are very few conditions that are directly associated with
glycolysis.
• It is a central energy yielding pathway, any defects are life threatening.
Most common disorder is Pyruvate kinase deficiency:
• The normal, mature erythrocyte lacks mitochondria and is completely dependent on

glycolysis for production of ATP.
• ATP is required to meet the metabolic needs of the red blood cell, and to fuel the
pumps necessary for the maintenance of the biconcave, flexible shape of the cell,
which allows it to squeeze through narrow capillaries.
• The premature death and lysis of red blood cells results in hemolytic anemia.
• Symptoms of PK deficiency is mild to severe chronic hemolytic anemia, icterus,

fatigue, jaundice, and elevated bilirubin levels.
Slide 31
SB10 The severity on the disease depends both of the degree of enzyme deficiency and the extent to which the
RBCs compensate by synthesising increased levels of 2,3-Bisphosphoglycerate from 1,3
bisphosphoglycerate. 2,3-BPG facilitates the release of oxygen from Hb into the tissue
Oxidative decarboxylation of pyruvate:
• If oxygen is available or in cells with mitochondria, pyruvate is further

oxidized to acetyl-CoA and carbon dioxide, producing more ATP
molecules.
• Oxidative decarboxylation of pyruvate by the enzyme pyruvate

dehydrogenase complex is an important reaction.
• Pyruvate dehydrogenase irreversibly converts pyruvate into acetyl CoA,

a major fuel for the TCA cycle and the building block for fatty acid
synthesis.
• +
Because there is only a limited amount of NAD in the cell, for glycolysis to
+
continue the NADH formed must be reoxidized to NAD by:
• oxidation of NADH via the respiratory chain (aerobic).
• NADH-linked conversion of pyruvate to lactate (anaerobic).
Reduction of pyruvate to lactate
• Lactate, formed by the action of lactate dehydrogenase, is the final product of

anaerobic glycolysis in eukaryotic cells.
• The formation of lactate is the major fate for pyruvate in lens and cornea of
the eye, kidney medulla, testes, leukocytes and red blood cells, because
these are all poorly vascularized and/or lack mitochondria.
SB9
• 1. Lactate formation in muscle: Exercising skeletal muscle, results in an

elevated NADH/NAD+ ratio, favouring reduction of pyruvate to lactate.
• Therefore, during intense exercise, lactate accumulates in muscle, causing a

drop in the intracellular pH, potentially resulting in cramps.
• Much of this lactate eventually diffuses into the bloodstream and can be used
by the liver to make glucose.
Slide 34
SB9 The direction of the LDH reaction depends on the intracellular concentration of pyruvate and lactate as
well as the concentrations of NADH/NAD+.
In liver and heart, due to the availability of NAD+, lactate is oxidised to pyruvate. In liver, pyruvate is
either converted to glucose (via gluconeogenesis) or oxidised by TCA cycle to CO2. In heart, pyruvate is
oxidised by TCA cycle.
SB8
Lactic acidosis:
• Elevated concentrations of lactate in the plasma, termed lactic acidosis, occur when
there is a collapse of the circulatory system, such as in
• myocardial infarction, pulmonary embolism, and uncontrolled hemorrhage, or
when an individual is in shock.
• The failure to bring adequate amounts of oxygen to the tissues results in impaired
oxidative phosphorylation and decreased ATP synthesis.
• To survive, the cells use anaerobic glycolysis as a backup system for generating
ATP, producing lactic acid as the end product.
• The excess oxygen required to recover from a period when the availability of oxygen
has been inadequate is termed as oxygen debt.
• In many clinical situations, measuring the blood levels of lactic acid allows the rapid,
early detection of oxygen debt in patients and the monitoring of their recovery.
Slide 35
SB8 Production of small amounts of ATP can be life-saving during the period required to re-establish
adequate blood flow to the tissues.
Energy yield
1. Aerobic glycolysis: A net gain of two ATP and two NADH per molecule of
glucose during glycolysis.
• Pyruvate enters citric acid cycle for further oxidation.
• NADH and FADH2 produced during glycolysis and citric acid cycle
enters oxidative phosphorylation.
• Total net production of 32 ATPs during cellular respiration.
2. Anaerobic glycolysis: A net gain of two molecules of ATP are generated for
each molecule of glucose converted to two molecules of lactate.
• There is no net production or consumption of NADH.
• Citric acid cycle and oxidative phosphorylation does not occur.
GLUCONEOGENESIS
SHARI R. BABU
shari.babu@unza.zm
Shari Babu
OVERVIEW OF GLUCONEOGENESIS
• Some tissues, such as the brain, red blood cells, kidney medulla, lens and cornea of
the eye, testes, and exercising muscle, require a continuous supply of glucose as a
metabolic fuel.
• Liver glycogen, an essential postprandial source of glucose, can meet these needs
for only 10–18 hours in the absence of dietary intake of carbohydrate.
• The brain uses almost 70% of the total glucose produced by liver during normal
fasting.
• During a prolonged fast, however, hepatic glycogen stores are depleted, and glucose
is formed from non-carbohydrate precursors.
• Gluconeogenic precursors include intermediates of glycolysis and the tricarboxylic

acid (TCA) cycle, as well as glycerol, lactate, and the a-keto acids obtained from the
transamination of glucogenic amino acids.
Shari Babu
ALANINE LACTATE OXALOACETATE
Aspartate, α-
Ketoglutarate,
PYRUVATE Fumarate, Succinyl-CoA
GLUCONEOGENESIS
GLUCOGENIC
AMINO ACIDS
DIHYDROXYACETONE PHOSPHATE
GLYCEROL FATTY ACID
TRIACYLGLYCEROL
Shari Babu
• The formation of glucose does not occur by a simple reversal of glycolysis.
• Instead, glucose is synthesized by a special pathway that requires both

mitochondrial and cytosolic enzymes.
• During an overnight fast, approximately 90% of gluconeogenesis occurs in the

liver, with the kidneys providing 10% of the newly synthesized glucose
molecules.
• However, during prolonged fasting, the kidneys become major glucose-

producing organs, contributing an estimated 40% of the total glucose
production.
Shari Babu
REACTIONS UNIQUE TO GLUCONEOGENESIS
• Seven out of the ten glycolytic reactions are reversible and use the same
enzymes in the synthesis of glucose from pyruvate via gluconeogenesis.
• However, three of the glycolytic reactions are irreversible and must be

circumvented by four alternate reactions that energetically favors the synthesis
of glucose.
• The three irreversible reactions that needs to be circumvent are the glycolytic
reactions catalyzed by hexokinase, phosphofructokinase-1 and pyruvate
kinase.
Shari Babu
Pink arrows Blue arrows indicate
indicate glycolytic the gluconeogenic
pathway pathway
Shari Babu
FORMATION OF PHOSPHOENOLPYRUVATE (PEP) FROM PYRUVATE
• Reversal of the reaction catalyzed by pyruvate kinase in glycolysis involves

two reactions.
1. Carboxylation of pyruvate: Pyruvate Carboxylase catalyzes an ATP-

requiring reaction in which the vitamin biotin is the coenzyme:
Pyruvate + HCO3- + ATP  Oxaloacetate + ADP + Pi
• Pyruvate carboxylase is allosterically activated by acetyl CoA and inhibited by

ADP.
Shari Babu
Transport of oxaloacetate (OAA) to the cytosol
• OAA must be converted to PEP for gluconeogenesis to continue.
• The enzyme that catalyzes this conversion is found in both the mitochondria
and the cytosol in humans.
• The PEP that is generated in the mitochondria is transported to the cytosol

by a specific transporter, whereas for PEP to form in the cytosol it requires
the transport of OAA from the mitochondria to the cytosol.
• OAA is unable to directly cross the inner mitochondrial membrane; it must

first be reduced to malate by mitochondrial malate dehydrogenase.
• Malate can be transported from the mitochondria to the cytosol, where it is

reoxidized to oxaloacetate by cytosolic malate dehydrogenase as NAD is +
reduced.
Shari Babu
CYTOSOL MITOCHONDRIAL
MATRIX
NAD+ NADH
MALATE OXALOACETATE
Malate
Dehydrogenase
Malate
Transporter
2. Decarboxylation of cytosolic oxaloacetate: Oxaloacetate is decarboxylated
and phosphorylated to PEP in the cytosol by PEP-carboxykinase. The reaction
is driven by hydrolysis of GTP.
Oxaloacetate + GTP  Phosphoenolpyruvate + GDP + CO2
• Then, PEP is acted on by the reactions of glycolysis running in the reverse

direction until it becomes fructose 1,6-bisphosphate.
• PEPCK is inhibited by ADP.
Shari Babu
DEPHOSPHORYLATION OF FRUCTOSE 1,6-BISPHOSPHATE
• Hydrolysis of fructose 1,6-bisphosphate by fructose 1,6-bisphosphatase

bypasses the irreversible phosphofructokinase-1 reaction.
Fructose 1, 6-bisphosphate + H2O  Fructose 6-phosphate + Pi
• This reaction is an important regulatory site of gluconeogenesis.
• Fructose 1,6-bisphosphatase is inhibited by elevated levels of AMP while high

levels of ATP and citrate stimulate gluconeogenesis, an energy-requiring
pathway.
• Fructose 1,6-bisphosphatase, found in liver and kidney, is also inhibited by

fructose 2,6-bisphosphate.
Shari Babu
SB7
• Fructose-2,6-bisphosphate is synthesized & degraded by a bi-functional

enzyme (PFK2/F2,6BPase) that includes 2 catalytic domains:
Phosphofructokinase-2 (PFK2) domain catalyzes:

Fructose-6-phosphate + ATP  Fructose-2,6-bisphosphate + ADP
Fructose-2, 6 Bisphosphatase (F2,6BPase) domain catalyzes:

Fructose-2,6-bisphosphate + H2O  Fructose-6-phosphate + Pi
Shari Babu
Slide 12
SB7 Phosphofructokinase 1 (PFK), which catalyses the phosphorylation of fructose-6-phosphate to

fructose-1,6- bisphosphate, is a key regulatory step in the glycolysis.
When glucose level is low, glucagon is released into the bloodstream, triggering a cAMP signal cascade.
In the liver Protein kinase A inactivates the PFK-2 domain of the bifunctional enzyme via phosphorylation.
The F-2,6-BPase domain is then activated which lowers fructose 2,6-bisphosphate (F-2,6-BP) levels.
Because F-2,6-BP normally stimulates phosphofructokinase-1(PFK1) and inhibits F-1,6BPase, the decrease
in its concentration leads to the inhibition of glycolysis and the stimulation of gluconeogenesis,
respectively.
GLUCAGON AND/OR EPINEPHRINE
cAMP-dependent Protein Kinase

(PKA)
+ Phosphorylation -
P
PFK-2 PFK-2
F-2,6-BPase F-2,6-BPase P
- +
Fructose-2, 6-Bisphosphate Fructose-6-Phosphate +

Pi
• Decreased levels of F-2,6-BP means inhibition of F-1,6-BPase is lifted.

• This increases the rate of Gluconeogenesis.
INSULIN
Protein Phosphatase-1
Dephosphorylation
+ -
P
P
F-2,6-BPase F-2,6-BPase
P
PFK-2 PFK-2
P
- +
Fructose-6-Phosphate + ATP Fructose-2, 6-

Bisphosphate + ADP
• Increased levels of F-2,6-BP activates PFK-1.

• This increases the rate of Glycolysis.
In presence of glucagon or PKA action In presence of insulin,
epinephrine, cAMP levels glucose will enter cells.
are increased.
F2,6BPase The levels of fructose-6-

cAMP-dependent protein phosphate will increase.
kinase (PKA) is activated.
PFK2 Insulin decreases the
levels of cAMP.
PKA activates F2,6BPase
domain via
phosphorylation. Activate Insulin also activates
Inactive
protein phosphatase-1
(PP1).
Once activated,
F2,6BPase will breakdown PP1 dephosphorylates
F2,6BP and lift its inhibition and activates PFK2.
PP1 action
on F1,6BPase.
This results in increased

PKA inactivates PFK2 via levels of F26BP which
phosphorylation preventing activates PFK1.
formation of F2,6BP.
Also, PP1
dephosphorylates and
Gluconeogenesis inactivates F26BPase.
stimulated.
Glycolysis is stimulated.
SB8
DEPHOSPHORYLATION OF GLUCOSE 6-PHOSPHATE
• Hydrolysis of glucose 6-phosphate by glucose 6-phosphatase bypasses the

irreversible hexokinase reaction.
Glucose 6-phosphate + H2O  Glucose + Pi
• Release of free glucose requires two proteins: glucose 6-phosphate

translocase, which transports glucose 6-phosphate across the ER membrane,
and the ER enzyme, glucose 6-phosphatase, which removes the phosphate,
producing free glucose.
Shari Babu
Slide 16
SB8 The deficiency of any one of the four unique gluconeogenic enzymes i.e., pyruvate carboxylase,
phoshoenolpyruvate carboxykinase, fructose 2,6-bisphosphatase and glucose-6-phosphatase, can lead to
hypoglycemia during periods of fasting. Especially overnight fasting when the liver glycogen reserves
have been depleted.
SB9
HORMONAL REGULATION OF GLUCONEOGENESIS
• Glucagon lowers the level of fructose 2,6-bisphosphate, resulting in activation

of fructose 1,6-bisphosphatase and inhibition of PFK-1, thus favouring
gluconeogenesis.
• Glucagon elevates cAMP level and cAMP-dependent protein kinase activity,

which stimulates the conversion of pyruvate kinase to its inactive
(phosphorylated) form.
• This decreases the conversion of PEP to pyruvate, which has the effect of
diverting PEP to the synthesis of glucose.
• Glucagon increases the transcription of PEP-carboxykinase gene, increasing

the levels of the enzyme.
• Insulin increases the activity of phosphofructokinase-1, pyruvate kinase and

the levels of F-2,6-BP.
Shari Babu
Slide 17
SB9 So in the liver and kidneys, glycolysis and gluconeogenesis do not occur simultaneously.
But it is possible for gluconeogenesis to occur in the liver while glycolysis occurs in extra-hepatic tissues.
CLINICAL SIGNIFICANCE
• In the absence of glucose-6-phosphatase, gluconeogenesis is impaired, and

this results in fasting hypoglycemia. This can occur in Von Gierke’s disease
when the enzyme is deficient.
• Diabetes is either the result of impaired insulin production or decreased

insulin sensitivity.
• Insulin is an inhibitor of gluconeogenesis. In the absence of insulin,
gluconeogenesis occurs at a rapid rate, exacerbating hyperglycemia.
• Alcoholism can induce hypoglycemia.

• Alcohol is oxidized to acetaldehyde and then to acetate in the liver. These
reactions increase the level of NADH.
• The high levels of NADH favors the formation of lactate from pyruvate.
• This lowers the level of pyruvate that can enter gluconeogenesis, which
causes hypoglycemia.
• As a result, heavy ethanol consumption can lead to both lactic acidosis
and hypoglycemia.
SB10
The Cori Cycle
• Lactate is formed by active skeletal muscle when the rate of glycolysis

exceeds the rate of oxidative metabolism.
• Lactate is readily converted into pyruvate by the action of lactate

dehydrogenase.
• During anaerobic glycolysis in skeletal muscle, pyruvate is reduced to lactate

by lactate dehydrogenase (LDH).
• Lactate produced by the LDH reaction is released to the blood stream and
transported to the liver where it is converted to glucose.
• The glucose is then returned to the blood for use by muscle as an energy
source and to replenish glycogen stores. This cycle is termed the Cori cycle.
Shari Babu
Slide 19
SB10 During vigorous muscular activity, epinephrine is released and it stimulates hepatic gluconeogenesis.
Cori Cycle
Shari Babu
CITRIC ACID CYCLE
SHARI BABU
shari.babu@unza.zm
Biomedical Importance
• The citric acid cycle (Krebs cycle, tricarboxylic acid cycle) is the second stage of cellular
respiration.
• It is a series of reactions in mitochondria that oxidize acetyl residues (as acetyl-CoA).
• The acetyl groups are fed into the citric acid cycle, which enzymatically oxidizes them
to CO2. The energy released by oxidation is conserved in the reduced electron carriers
NADH and FADH2.
• It is the final common pathway for the aerobic oxidation of carbohydrate, lipid, and
protein because glucose, fatty acids, and most amino acids are metabolized to acetyl-
CoA or intermediates of the cycle.
• It also produces important precurors that form building blocks of many other
molecules.
Cellular Aerobic Respiration
Catabolism of carbohydrates, lipids and amino acids generate
ACETYL-CoA
FATTY
PYRUVATE ACID
KETOGENIC KETONE
AMINO BODIES
ACIDS
ACETYL-CoA
Transport and Production of Acetyl-CoA
• Pyruvate generated in the cytoplasm by glycolysis must be transported

across the inner mitochondrial membrane via a pyruvate/H+ symport.
• This transport uses some of the energy stored in the mitochondrial inner
membrane electrical potential gradient.
• Pyruvate derived from glucose and other sugars by glycolysis, is oxidized to

acetyl-CoA and CO2 by the pyruvate dehydrogenase (PDH) complex,
located in the mitochondria of eukaryotic cells and in the cytosol of
prokaryotes.
The Pyruvate Dehydrogenase Complex Consists of Three
Distinct Enzymes
• The PDH complex contains three enzymes—pyruvate dehydrogenase (E1),

dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide
dehydrogenase (E3).
The Pyruvate Dehydrogenase Complex Requires Five

Coenzymes
• They are thiamine pyrophosphate (TPP), flavin adenine dinucleotide

(FAD), coenzyme A, nicotinamide adenine dinucleotide (NAD), and
lipoate.
• The active site of E1 has bound TPP, E2 is the point of connection for the
prosthetic group lipoate, and E3 has bound FAD.
SB1
SB2
SB3
CoASH
ACETYL-CoA
PYRUVATE TPP Acetyl-Lipoamide
E1
Lipoamide
E2 Reduced
Hydroxyethyl
CO2 TPP
Lipoamide
Oxidized
FAD
E3
FADH2
E1 – Pyruvate dehydrogenase NADH + H+
E2 – Dihydrolipoamide acetyltransferase NAD+
E3 – Dihydrolipoamide dehydrogenase
Slide 7
SB1 In the first step, E1 (pyruvate dehydrogenase) catalyzes oxidative decarboxylation of pyruvate. One carbon atom is removed from pyruvate as CO2.
The resulting 2-carbon molecule, hydroethyl is bound to TPP.
SB2 Next step is catalyzed by E2 (dihydrolipoamide acetyltransferase), which transfers the hydroethyl group from TPP to the oxidized form of lipoamide.
The acetyl group is then transferred to free coenzyme A to form acetyl-CoA and reduced lipoamide.
SB3 Finally, FAD-dependent E3 (dihydrolipoamide dehydrogenase) reoxidizes the lipoyl group E2. The electrons are accepted by FAD to form FADH2,
which is then donated to NAD+ to form NADH
• The overall reaction catalyzed by the pyruvate dehydrogenase complex is
an oxidative decarboxylation, an irreversible oxidation process in which
the carboxyl group is removed from pyruvate as a molecule of CO2 and the
two remaining carbons become the acetyl group of acetyl-CoA.
• Deficiencies of thiamine can cause serious central nervous system
problems.
• This is because brain cells are unable to produce sufficient ATP (via the TCA
cycle) if the PDH complex is inactive.
• Beriberi, a disease that results from thiamine deficiency, is characterized

by loss of neural function.
• Wernicke-Korsakoff, an encephalopathy-psychosis syndrome due to

thiamine deficiency, may be seen with alcohol abuse.
• An elevated level of pyruvate in the blood is often an indicator of defects in

pyruvate oxidation due to one of these causes.
Regulation of acetyl-CoA production by the PDH complex
• When AMP, CoA, and NAD+ accumulate, they allosterically activate the
pyruvate dehydrogenase complex.
• The PDH complex is inhibited by reversible phosphorylation of E1.

• A specific protein kinase called PDH kinase, phosphorylates and
inactivates E1.
• PDH kinase is activated by ATP, acetyl-CoA and NADH.
• PDH kinase is inhibited by pyruvate.
• A specific phosphatase called PDH phosphatase, removes the

phosphoryl group by hydrolysis and thereby activates E1.
• Calcium is a strong activator of PDH phosphatase
Pyruvate dehydrogenase deficiency:
• A deficiency in the E1 component of the PDH complex is the most common

biochemical cause of congenital lactic acidosis.
• This enzyme deficiency results in accumulation of pyruvate, causing it to be

converted to lactic acid.
• This causes problems for the brain, which relies on the TCA cycle for most of its
energy and is particularly sensitive to acidosis.
• Symptoms are variable and include neurodegeneration, muscle spasticity and, in the
neonatal onset form, early death.
• The E1 defect is X-linked it affects both males and females and is classified as X-
linked dominant.
• There is no proven treatment for PDH deficiency: however, dietary restriction of

carbohydrate and supplementation with TPP may reduce symptoms in select
patients.
Reactions of the Citric Acid Cycle
Acetyl-CoA
CoASH
Oxaloacetate Citrate
NADH + H+
NAD+
Malate Isocitrate
KERBS NAD+
CYCLE NADH + H+ + CO2
Fumarate α-Ketoglutarate
NAD+ + CoASH
FADH2
FAD NADH + H+ + CO2

Succinate Succinyl
-CoA
GTP/ATP GDP/ADP
The Citric Acid Cycle Has Eight Steps
1. Formation of Citrate: The first reaction of the cycle is the condensation of

acetyl-CoA with oxaloacetate to form citrate, catalyzed by citrate
synthase:
• The CoA liberated in this reaction is recycled to participate in the oxidative

decarboxylation of another molecule of pyruvate by the PDH complex.
2. Formation of Isocitrate via cis-Aconitate: Citrate is isomerized to
isocitrate by the enzyme aconitase.
• Fluoroacetate is an inhibitor of TCA cycle.

• Fluoroacetate is first converted to fluorocitrate by citrate synthase.
• Fluorocitrate is a potent inhibitor of aconitase.
SB4
Slide 14
SB4 Fluoroacetate is found in a range of plant species. Some flourinated compounds such as in anticancer agents, pesticides, and industrial chemicals can
be metabolized to fluoroacetate.
3. Oxidation of Isocitrate to α-Ketoglutarate and CO2: In the next step,
isocitrate dehydrogenase catalyzes oxidative decarboxylation of isocitrate
to form α-ketoglutarate.
• This is one of the rate-limiting steps of the TCA cycle. The enzyme is
allosterically activated by ADP and Ca , and is inhibited by ATP and NADH.
2+
4. Oxidation of α-Ketoglutarate to Succinyl-CoA and CO2: The next step is
another oxidative decarboxylation, in which α-ketoglutarate is converted
to succinyl-CoA and CO2 by the action of the α-ketoglutarate
dehydrogenase complex.
• α-Ketoglutarate dehydrogenase is nearly identical to PDH in terms of

reactions catalysed and its structural features.
• TPP, lipoid acid, CoASH, FAD and NAD+ are required.
• α-Ketoglutarate dehydrogenase complex is inhibited by its products, NADH

and succinyl CoA, and activated by Ca .
2+
5. Conversion of Succinyl-CoA to Succinate: Succinate thiokinase (succinyl
CoA synthetase) cleaves the high-energy thioester bond of succinyl CoA.
This reaction is coupled to phosphorylation of guanosine diphosphate
(GDP) to guanosine triphosphate (GTP).
• GTP formed is used for mitochondrial synthesis of proteins, RNA and DNA.
• The GTP formed can donate its phosphoryl group to ADP to form ATP, in a
reversible reaction catalyzed by nucleoside diphosphate kinase.
6. Oxidation of Succinate to Fumarate: The succinate formed from succinyl-
CoA is oxidized to fumarate by the flavoprotein succinate dehydrogenase:
• In eukaryotes, succinate dehydrogenase is tightly bound to the inner

mitochondrial membrane, and is the only enzyme of the citric acid cycle that
is membrane-bound.
• Succinate dehydrogenase is activated by ATP and succinate.
• Malonate, an analog of succinate, is a strong competitive inhibitor of

succinate dehydrogenase and the enzyme is also inhibited by oxaloacetate.
7. Hydration of Fumarate to Malate: The reversible hydration of fumarate
to L-malate is catalyzed by fumarase.
8. Oxidation of Malate to Oxaloacetate: In the last reaction of the citric acid
cycle, NAD+-linked L-malate dehydrogenase catalyzes the oxidation of L-
malate to oxaloacetate:
• Products of one turn of the citric acid cycle. Three NADH, one FADH2, one
GTP (or ATP), and two CO2 are released in oxidative decarboxylation
reactions.
• The three NADH, and one FADH2 enters into the electron transport chain
and drives ATP synthesis via oxidative phosphorylation.
• The cycle generates the equivalent of 10 ATPs from one acetyl-CoA

(3 NADH = 7.5 ATPs, 1 FADH2 = 1.5 ATPs, 1 ATP (GTP) directly)
Arsenic Poisoning
• Arsenic poisoning” is primarily due to inhibition of enzymes that require

lipoic acid as a coenzyme, including E2 of the PDH complex and α-
ketoglutarate dehydrogenase.
• Arsenic forms a stable complex with the thiol (–SH) groups of lipoic acid,
making it unavailable to serve as a coenzyme.
• When it binds to lipoic acid in the PDH complex, pyruvate (and consequently
lactate) accumulates.
• Like pyruvate dehydrogenase deficiency, this particularly affects the brain,

causing neurologic disturbances and death.
Citric Acid Cycle Source Of Biosynthetic Intermediates
• α-ketoglutarate simple transamination is converted to glutamate, and in

nervous tissues converted to the neurotransmitter, γ-aminobutyrate
(GABA).
• Oxaloacetate is transaminated to aspartate, the precursor of asparagine as

well as purine and pyrimidine bases.
• Oxaloacetate is converted to phosphoenolpyruvate, key intermediate of

gluconeogenesis.
• Succinyl-CoA is a central intermediate in the synthesis of the porphyrin ring

of heme groups, which serve as oxygen carriers (in hemoglobin and
myoglobin) and electron carriers (in cytochromes).
• Citrate exported into cytosol is converted to oxaloacetate and acetyl-CoA,

precursor for fatty acid and cholesterol synthesis.
GLUCOSE
Acetyl-
CoA
Oxaloac FATTY ACID

Citrate SYNTHESIS &
ASPARTATE etate
CHOLESTEROL
Malate Isocitrate
GLUTAMATE
Fumarat
α-KG
e
GABA
Succinat Succinyl
e -CoA
HEME
PENTOSE PHOSPHATE
PATHWAY
SHARI R. BABU
DEPARTMENT OF PHYSIOLOGICAL SCIENCES
SCHOOL OF MEDICINE
shari.babu@unza.zm
Pentose Phosphate Pathway
Also known as:

• Hexose monophosphate shunt
• Phosphogluconate pathway
• It occurs in the cytosol.

One fate of Glucose -6-
Phosphate is the Pentose
Phosphate Pathway.
The pentose pathway is a shunt.
• The pathway begins with the glycolytic intermediate

glucose 6-P.
• It shunts the carbons back to glycolytic pathway via the

products of the pentose pathway, i.e., glyceraldehyde 3-
phosphate and fructose 6-phosphate.
• It is for this reason that the pentose phosphate pathway is

often referred to as a shunt.
What is the importance of pentose
phosphate pathway?
• The pathway yields:
• NADPH required for the synthesis of fatty acids, sterols and

deoxyribonucleotides. Reduction of oxidized glutathione
needed for cellular antioxidant defense.
• ribose 5-phosphate required for synthesis of nucleic acid

(RNA and DNA) and cofactors (CoA, FAD, NAD+/
NADP+).
• There are minor mechanisms of generating NADPH, all of
which occur in the mitochondria.
• The important enzymes in these processes are: NADP-linked

isocitrate dehydrogenase and
SB2 NADP-linked glutamate
SB3
dehydrogenase.
• NADP-malic
SB1 enzyme is present in the cytosol.
SB8
Slide 7
SB1 NADP-malic enzyme catalyzes the oxidation of malate to pyruvate and carbon dioxide and, with the formation of NADPH.
SB8 Occurs in the cytosol in humans

SB2 NADP-isocitrate dehydrogenase catalyses conversion of isocitrate to alpha-ketoglutarate and NADPH is formed.
SB3 NADP-linked glutamate dehydrogenase catalyses the reversible conversion of glutamate to alpha-ketoglutarate and ammonia while reducing NADP
to NADPH.
Tissues with active Pentose Phosphate Pathway
• PPP occurs in the cytosol.
OXIDATIVE PHASE
• It can be divided into two phases.
• NADPH + H+ is formed from two

separate reactions.
• The glucose 6-phosphate

dehydrogenase (G6PD) reaction is
the rate limiting step and is
irreversible.
NON-OXIDATIVE PHASE
• Cells have a greater need for

NADPH than ribose 5-phosphate.
SB9
SB11
SB12
Slide 12
SB9 Transketolase enzyme (TPP as co-factor) accepts 2-carbon fragment from the ketose sugar, xylulose and transfers it to Ribose (an aldose) to form
sedoheptulose 7-P (ketose).
Xylulose with the loss of 2-carbon is converted to glyceraldehyde 3-P (aldose).
SB11 Transaldolase catalyses the transfer of a 3-carbon fragment from sedoheptulose 7-P to glyceraldehyde 3-P (aldolase sugar).
SB12 Transketolase transfer of a 2-carbon fragment from xylulose-5-P to the aldose erythrose-4-phosphate, forming fructose 6-phosphate and
glyceraldehyde-3-P.
• The reactions of the non-oxidative phase of the
pentose phosphate pathway are readily reversible.
• The non-oxidative phase can regulate the rate of

glycolysis or gluconeogenesis by providing F6P
and G3P, depending on the metabolic needs of the
cell or tissue.
When rapidly dividing cells require more ribose 5-
phosphate than NADPH.
When the need for NADPH and ribose 5-phosphate is
balanced.
When more NADPH is needed than ribose 5-phosphate.
When both NADPH and ATP is needed.
Regulation of the Pentose Pathway
• Glucose 6-phosphate dehydrogenase is the regulatory enzyme.
• High levels of NADP+ allosterically stimulating G6PD and increases the

flux of glucose 6-phosphate through the pentose phosphate pathway.
• When the demand for NADPH slows, the level of NADP+ drops, the
pentose phosphate pathway slows, and glucose 6-phosphate is instead used
to fuel glycolysis.
• The synthesis of glucose 6-phosphate dehydrogenase is induced by the

increased insulin level after a high carbohydrate meal.
ROLE OF PPP IN OBESITY INDUCED INSULIN
RESISTANCE
• Adipose tissue macrophages that surround dead adipocytes cause
obesity-induced inflammation and secrete pro-inflammatory cytokines.
• Pro-inflammatory cytokines released include tumor necrosis factor α

(TNF-α) and interleukin-1β (IL-1β)
• Pro-inflammatory macrophages show increased glycolysis and PPP

which generates more energy and NADPH to trigger inflammatory
response and secretes pro-inflammatory cytokines.
• Pro-inflammatory cytokines increases the level of G6PD in adipocytes

which increases the rate of PPP.
• This causes adipocytes to release more cytokines which further

stimulates inflammatory response.
• This vicious cycle promotes obesity induced insulin resistance, resulting
in severe Type 2 Diabetes Mellitus
• TNF-α and IL-1β bring about insulin resistance by altering the

insulin receptor signaling pathway.
• Pro-inflammatory cytokines also promote lipolysis in adipocytes,

leading to elevated levels of circulating free fatty acids.
• In the skeletal muscle, FFAs reduce insulin-stimulated glucose intake,

resulting in skeletal muscle insulin resistance.
ROLE OF PPP IN INSULIN SECRETION
• Insulin is stored in granules and released via exocytosis from

pancreatic β-cells in response to glucose.
• NADPH converts oxidized glutathione to reduced glutathione

which brings about insulin granule exocytosis.
• Inhibition of G6PD and 6PGD not only blocks glucose stimulated

insulin secretion but also increases oxidative stress and β-cell
apoptosis.
• Patients with G6PD deficiency show decreased insulin secretion.

ROLE OF PPP IN CANCER
• Tumor suppressor p53 regulates cell division by preventing cells
from dividing uncontrollably.
• When DNA is damaged this protein determines whether the DNA

can be repaired or if the cell will undergo apoptosis.
• p53 directly binds to G6PD and inhibits the enzyme, whereas

mutant p53 fails to inhibit G6PD in cancer cells.
• Up-regulation of G6PD provides cells with R5P for

nucleotide synthesis as well as NADPH for biosynthesis and
maintaining redox homeostasis.
• Suppression of G6PD leads to intracellular oxidative stress,

making cancer cells sensitive to chemotherapy.
ROLE OF PPP IN ANTIOXIDANT
DEFENSE
• What is glutathione?
• Why is it important?
• How is it related to NADPH?
Glutathione is a tripeptide
composed of glutamate,
cysteine, glycine.
Reduced glutathione
(GSH) maintains the
normal reduced state of
the cell.
Reduced glutathione
(GSH)
Glutathione Functions -1
• It serves as a reductant.
• Involved in metabolism of drugs making them water soluble.
• Cofactor in some enzymatic reactions.

– rearrangement of protein disulfide bonds.
Glutathione Functions -2
• The sulfhydryl of GSH is used to reduce peroxides (ROS) formed

during oxygen transport.
– Reactive oxygen species (ROS) damage macromolecules (DNA, RNA,
and protein) and ultimately lead to cell death.
• The resulting oxidized form of GSH is two molecules linked by a

disulfide bridge (GSSG).
• The enzyme glutathione reductase uses NADPH as a cofactor to

reduce GSSG back to two molecules of GSH.
• Thus, the pentose pathway is linked to the supply of adequate amounts

of GSH.
Reduction of hydrogen peroxide
• Hydrogen peroxide is one of the reactive oxygen species (ROS) that are formed from the
partial reduction of molecular oxygen.
• These compounds are formed continuously as by-products of aerobic metabolism,

through reactions with drugs and environmental toxins, or when the level of antioxidants
is diminished, all creating the condition of oxidative stress.
• The highly reactive oxygen intermediates can cause serious chemical damage to DNA,
proteins, and unsaturated lipids, and can lead to cell death.
• These ROS have been implicated in several pathologic processes, including cancer,
inflammatory disease, and aging.
• During normal detoxification, H2O2 is converted to H2O and O2 by catalase or H2O2 is

converted to H2O by glutathione peroxidase utilizing reduced glutathione.
Glutathione and Erythrocytes
• GSH is extremely important particularly in the highly

oxidizing environment of the red blood cell.
• Mature RBCs have no mitochondria and are totally

dependent on NADPH from the pentose phosphate pathway
to regenerate GSH via glutathione reductase.
• In fact, as much as 10% of glucose consumption, by

erythrocytes, is mediated by the pentose phosphate
pathway.
SB13
Role of G6PD in red blood cells
• Diminished G6PD activity impairs the ability of the cell to form the NADPH
that is essential for the maintenance of the reduced glutathione pool.
• Glutathione also helps maintain the reduced states of sulfhydryl groups in

proteins, including hemoglobin.
• Oxidation of those sulfhydryl groups by H2O2 leads to the formation of cross-

linked complexes of denatured hemoglobin that form insoluble masses (called
Heinz bodies) that attach to the red cell membranes.
• Additional oxidation of membrane proteins causes the red cells to be rigid, and
they are removed from the circulation by macrophages in the spleen and liver.
• GSH is essential for normal RBC structure and keeping hemoglobin iron in Fe2+
state.
Slide 30
SB13 Disulfide bridges exists only in proteins that are located outside the cells. Inside the cell the cysteine -SH gro0ups are kept in their recued form.
GLUCOSE 6-P DEHYDROGENASE DEFICIENCY
• Glucose 6-phosphate dehydrogenase (G6PD) deficiency is an

inherited disease characterized by hemolytic anemia.
• G6PD deficiency is X-linked, and is the most common disease-

producing enzyme abnormality in humans.
• In addition to hemolytic anemia, a clinical manifestation of G6PD

deficiency is neonatal jaundice appearing 1–4 days after birth.
• The jaundice, which may be severe, typically results from

increased production of unconjugated bilirubin.
• People with this disorder are not normally anemic and
display no evidence of the disease until the red cells are
exposed to an oxidant or stress.
Drugs that can precipitate this reaction:

• Antimalarial agents (primaquine)
• Sulfonamides (antibiotic)
• Aspirin
• Non-steroidal anti-inflammatory drugs (NSAIDs)
OXIDATIVE PHOSPHORYLATION
SHARI BABU
shari.babu@unza.zm
STAGE 3 OF AEROBIC RESPIRATION: ELECTRON TRANSPORT CHAIN AND
SHARI R BABU
ELECTRON TRANSPORT CHAIN
The set of proteins and small molecules involved in the

orderly sequence of electron transfer to oxygen within
the inner mitochondrial membranes.
SHARI R BABU
Oxidative Phosphorylation at the respiratory chain level,

is the process in which ATP is formed as a result of
transfer of the electrons to O2 by a series of electron
carriers.
SHARI R BABU
• The intermediates of metabolic reactions donate electrons to specific
coenzymes—NAD+ and FAD—to form the energy-rich reduced coenzymes,
.
NADH and FADH2.
• These reduced coenzymes can, in turn, each donate a pair of electrons to a

specialized set of electron carriers.
• As electrons are passed down the electron transport chain (ETC), they lose
much of their free energy. Part of this energy can be captured and stored
by the production of ATP from ADP and inorganic phosphate (Pi).
• Electron transport and ATP synthesis by oxidative phosphorylation proceed

continuously in all tissues that contain mitochondria.
SHARI R BABU
• The components of the electron transport chain are located in the inner membrane.
• The inner mitochondrial membrane can be disrupted into five separate protein
complexes, called Complexes I, II, III, IV, and V.
• Complexes I–IV each contain part of the electron transport chain.
• Each complex accepts or donates electrons to relatively mobile electron carriers,

such as coenzyme Q and cytochrome c.
• Each carrier in the electron transport chain can receive electrons from an electron
donor, and can subsequently donate electrons to the next carrier in the chain.
• The electrons ultimately combine with oxygen and protons to form water. This
requirement for oxygen accounts for the greatest portion of the body’s use of
oxygen.
• Complex V catalyzes ATP synthesis and so is referred to as ATP synthase.

SHARI R BABU
SUBSTRATE SHUTTLES TRANSPORTING REDUCING EQUIVALENTS
• Two substrate transport shuttle employed in transporting reducing equivalents (e.g.,

electrons and protons) carried by NADH from cytosol to mitochondria, or reverse are:
glycerol-phosphate shuttle and malate-aspartate shuttle.
• These shuttle systems are required since inner mitochondrial membrane is

impermeable to NADH.
Glycerol-phosphate shuttle
• NADH produced in the cytosol is used to reduce dihydroxyacetone phosphate to glycerol-

3-phosphate by cytosolic glycerol-3- phosphate dehydrogenase.
• Glycerol-3-phosphate enters the mitochondria and is oxidized back to dihydroxyacetone

phosphate by mitochondrial glycerol-3-phosphate dehydrogenase and FADH2 that is
oxidized by the ETC.
SHARI R BABU
INTERMEMBRANE SPACE
MITOCHONDRIAL
MITOCHONDRIAL
MEMBRANE
MEMBRANE
INNER
OUTER
NADH + H+ DHAP DHAP
FADH2
Glycerol-3-P dehydrogenase Glycerol-3-P dehydrogenase
NAD+ Glycerol-3- Glycerol-3-

FAD
phosphate phosphate
SHARI R BABU
Malate-Aspartate Shuttle
• NADH in the cytosol reduces oxaloacetate to malate-by-malate dehydrogenase,

malate then enters the mitochondria.
• This malate is then readily oxidized by mitochondrial malate dehydrogenase to

oxaloacetate and NADH that is then oxidized by the ETC.
• The resulting oxaloacetate undergoes a transamination reaction to form

aspartate by aspartate aminotransferase, which can be transported to the
cytosolic side.
• In the cytoplasm, aspartate is then transaminated to form oxaloacetate and the

cycle is restarted.
• This shuttle, in contrast with the glycerol 3-phosphate shuttle, is readily

reversible.
SHARI R BABU
MITOCHONDRIAL
CYTOSOL MITOCHONDRIAL MATRIX
MEMBRANE
Aspartate Aspartate
Transamination
Transamination
Oxaloacetate Oxaloacetate NADH
NADH + H+
+ H+
Malate dehydrogenase
NAD+ Malate Malate NAD+
SHARI R BABU
REACTIONS OF ELECTRON TRANSPORT CHAIN
• With the exception of coenzyme Q (ubiquinone), all members of this chain are
proteins.
• The pair of electrons carried by NADH along with two hydrogen atoms are
transferred to NADH dehydrogenase or NADH-ubiquinone oxidoreductase
(Complex I) complex embedded in the inner mitochondrial membrane.
• Complex I contains iron atoms paired with sulfur atoms to make iron–sulfur
clusters as well as FMN.
• The electrons are transferred one at a time via FMN and a series of FeS
clusters.
• During the transfer of electrons to CoQ by Complex I, four protons are

translocated across the inner membrane to the inter membrane space.
SHARI R BABU
• FADH2 transfers electrons to CoQ via FeS centers of Complex II (succinate
dehydrogenase).
• The amount of free-energy released during these reactions is insufficient

for proton pumping across the inner membrane.
• CoQ when it accepts electrons become reduced to ubiquinol (CoQH2).
• CoQ transfers electrons to Complex III (cytochrome c reductase or

cytochrome bc1).
• The remaining members of the electron transport chain are cytochromes.
• Each contains a heme group (a porphyrin ring plus iron). The heme iron is
reversibly converted from its ferric (Fe3+) to its ferrous (Fe2+) form as a
normal part of its function as a reversible carrier of electrons.
SHARI R BABU
• Electrons are passed along the chain from CoQ to cytochrome bc (Complex III) to
1
cytochrome c and to cytochrome c oxidase (Complex IV).
• The transfer of electrons through complex III results in pumping of 4 protons into the
inter membrane space.
• Cytochrome c is associated with the outer face of the inner membrane and, like CoQ,
is a mobile carrier of electrons.
• Cytochrome c oxidase complex (Complex IV) is the only electron carrier in which the
heme iron can react directly with O2.
• Cytochrome oxidase contains copper atoms that are required for this complex
reaction to occur.
• At this site, the transported electrons and O2, are brought together, and O2 is
reduced to water by reacting to 2 protons from the mitochondrial matrix and 2
protons are pumped into the intermembrane space.
SHARI R BABU
SB5
SB6
OUTER MITOCHONDRIAL MEMBRANE
INTERMEMBRANE SPACE ACIDIC H+

H+ H+
2H+ H+
4H+ 4H+ H+ H+
H+
H+
2e - 2e - e-
e-
FeS CoQ
+ + + + + + Cyt c + + + +
+ +
INNER
-
MITOCHONDRIAL
- I - - 2e - - - - - - IV - - - -
FeS
II
MEMBRANE
FMN
2e -
2e -
FAD FADH2
III e-
V
NADH NAD+ SUCCINATE FUMARATE H2 O
½ O 2 + 2 H+
NADH FADH2
KERBS BETA
CYCLE OXIDATION
BASIC
SHARI R BABU
MITOCHONDRIAL MATRIX
Slide 14
SB5 Because cytochrome c can only accept one electron at a time, the transfer of electron from CoQ to Cyt c through Complex III occurs twice.
SB6 When each elctron passes through Complex III 2H+ are pumped into the intermembrane space. So when the 2 electrons are transferred to Cyt c, a
total of 4H+ are pumped into the intermembrane space.
SITE-SPECIFIC INHIBITORS:
• Rotenone, amytal, piericidin (an antibiotic) and amobarbital block electron

transfer through Complex I.
• Complex II is inhibited by carboxin, thenoyltrifluoroacetone and malonate,

oxaloacetate.
• Doxorubicin (used in chemotherapy) interferes with electron transfer by

CoQ.
• Antibiotics like Antimycin A, myxothiazol and stigmatellin inhibit with

electron flow in Complex III. Dimercaprol (chelating agent) also is an
inhibitor.
• Cyanide, hydrogen sulfide, sodium azide and CO block electron flow

through Complex IV.
SHARI R BABU
SITE-SPECIFIC INHIBITORS:
• These inhibitors prevent the passage of electrons by binding to a

component of the chain, blocking the oxidation/reduction reaction.
• Incomplete reduction of oxygen to water produces reactive oxygen species

(ROS), such as superoxide (O2–•), hydrogen peroxide (H2O2) and hydroxyl
radicals (OH•).
• ROS damage DNA and proteins, and cause lipid peroxidation. Enzymes such
as superoxide dismutase (SOD), catalase, and glutathione peroxidase are
cellular defenses against ROS.
SHARI R BABU
• The chemiosmotic hypothesis (also known as the Mitchell hypothesis)
explains how the free energy generated by the transport of electrons by the
ETC is used to produce ATP from ADP + Pi.
• Proton pump: During electron transport the pumping of protons (H+)
across the inner mitochondrial membrane from the matrix to the
intermembrane space at Complexes I, III, and IV is coupled with the
synthesis of ATP.
• This process creates an electrical gradient and a pH gradient
(electrochemical gradient, proton gradient or proton motive force).
• The energy generated by this proton gradient is sufficient to drive ATP
synthesis.
SHARI R BABU
SB2
SB3
• ATP synthase: The enzyme complex ATP synthase (Complex V)
synthesizes ATP using the energy of the proton gradient
generated by the ETC.
• ATP synthase contains F0 and F1 domains which acts as F0 domain

rotational motor system.
• The chemiosmotic hypothesis proposes that protons re-enter

the matrix by passing through a channel in the membrane-
spanning domain F0.
F1 domain
• F0 rotation causes conformational changes in the F1 domain
that allow it to bind ADP + Pi and phosphorylate to ATP.
• Oligomycin binds to the F0 domain, closing the H channel,

+
preventing re-entry of protons into the mitochondrial matrix,

and thus preventing phosphorylation and release of ATP.
SHARI R BABU
Slide 18
SB2 As protons flow through F0, it becomes protonated and deprotonated repeatedly. This alternating ionization of F0 causes rotation.
SB3 For every 4 proton that re-enters, 1 ATP is synthesized.

INTERMEMBRANE SPACE ACIDIC H+

H+ H+ +
2H+ H
4H+ 4H+ H +
H+ + H+
H H+
2e - 2e - e-
e- Cyt
CoQ + + + + +
+ + + + + c
+ +
e- IV
INNER
-
MITOCHONDRIAL
MEMBRANE
- I - -
II
-
III - - - - - - - -
2e -
e-
SUCCINATE FUMARATE V
NADH NAD+ ½ O 2 + 2 H+ H2 O
NADH FADH2
BASIC
ADP + Pi ATP
H+ H+
KERBS H+
CYCLE BETA
OXIDATION
SHARI R BABU
Source: https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.electronicslovers.com%2F2018%2F05%2Fhydroelectric-power-site-selection- SHARI R BABU
key-components-how-it-works
SB1
SB4 ENERGETICS OF AEROBIC CELLULAR RESPIRATION
STAGE 1 GLUCOSE GLYCOLYSIS 2 PYRUVATE + 2 NADH + 2 ATP
2 ACETYL-CoA + 2 NADH
STAGE 2 SB8
2 X 3 NADH
KERBS
CYCLE
2 X 1 FADH2 2 GTP
8-10 NADH 8-10 X 2.5 20-25 ATP

STAGE 3 ETC &
2-4 FADH2 2-4 X 1.5 3-6 ATP
SB7
30-32 ATP
SHARI R BABU
Slide 21
SB1 In different literature the total ATP produced per molecule of glucose during cellular respiration varies from 32-38. This is because in some literature
they round off the number of ATPs produced per NADH from 2.5 to 3, and for FADH2 from 1.5 to 2 ATPs.
SB4 For every 4 proton that re-enters, 1 ATP is synthesized.

SB7 The 2 NADH formed in glucolysis is in the cutosol. Depending which shuttle system transports the electron of NADH into the mitochondrial matrix,
that is if glycerol phosphate shuttle delivers electrons viz FADH2 or the malate aspartate shuttle carries the electrons via NADH.
SB8 30-32 ATP from the breakdown of one glucose molecule is a high-end estimate, and the real yield may be lower. For instance, some intermediates
from cellular respiration may be channeled by the cell and used in other biosynthetic pathways, reducing the number of ATP produced.
• Electron transport is normally tightly coupled to ATP synthesis.
• When ADP levels are high in the mitochondria, the rate of oxygen
consumption also rises as electrons flow down the chain, and then
the rate of oxygen utilization falls when all the ADP has been
phosphorylated to ATP; a process called RESPIRATORY CONTROL.
• This mechanism ensures that electrons flow down the chain only
when ATP synthesis is needed.
• If the level of ATP is high and the ADP level is low, no electron
transport occurs, NADH and FADH2 build up, as does excess citrate,
and the citric acid cycle and glycolysis are inhibited.
SHARI R BABU
• Uncoupling proteins (UCP): UCPs occur in the inner mitochondrial
membrane of mammals, including humans.
• These carrier proteins create a “proton leak,” that is, they increase the
permeability of the inner membrane allowing protons to re-enter the
mitochondrial matrix without energy being captured as ATP.
• The energy is released as heat, and the process is called non-shivering
thermogenesis.
• UCP1, also called thermogenin, is responsible for the heat production
in the brown adipocytes of mammals.
• Brown fat uses almost 90% of its respiratory energy for thermogenesis
in response to cold in the neonate, and in hibernating animals.
• However, humans appear to have little brown fat (except in the
newborn), and UCP1 does not appear to play a major role in energy
balance.
SHARI R BABU
• When synthetic uncouplers such as 2,4-dintrophenol (DNP), dicumarol,
carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) is added to
cells, it stops ATP synthesis, but electron transport continues.
• DNP and other uncoupling agents are lipid-soluble small molecules that
can bind H+ ions and transport them across membranes (H+
ionophores).
• This prevents formation of a proton gradient. Since no proton gradient

forms, no ATP can be generated by oxidative phosphorylation.
• In high doses, aspirin and other salicylates uncouple oxidative phos-

phorylation.
SHARI R BABU
INTERMEMBRANE SPACE H+
2H+ H+
4H+ 4H+ H+ +
H H+
e- e-
e- Cyt e- ATP
CoQ H+
c synthase
INNER
I e- III IV
H+ UCP
II
MITOCHONDRIAL
MEMBRANE
e-
e-
SUCCINATE FUMARATE
NADH NAD+
½ O 2 + 2 H+ H2O
H+
NADH FADH2 H+ H+
H+ H
+
KERBS
CYCLE BETA
OXIDATION
SHARI R BABU
Inherited defects in oxidative phosphorylation
• Most of the proteins required for oxidative phosphorylation are coded for
by mtDNA and synthesized in mitochondria.
• Defects in oxidative phosphorylation are more likely a result of

alterations in mtDNA, which has a mutation rate about ten times greater
than that of nuclear DNA.
• Tissues with the greatest ATP requirement (for example, central nervous
system, skeletal and heart muscle, kidney, and liver) are most affected
by defects in oxidative phosphorylation.
• Most common clinical manifestations of defects in oxidative

phosphorylation are mitochondrial myopathies (seizures, hypotonia,
ophthalmoplegia, stroke-like episodes, muscular weakness),
cardiomyopathy.
SHARI R BABU
Mitochondria and apoptosis
• The process of apoptosis or programmed cell death may be initiated

through mitochondrial-mediated pathway by the formation of pores
in the outer mitochondrial membrane.
• These pores allow cytochrome c to leave the intermembrane space

and enter the cytosol.
• Once in the cytosol, cytochrome c, in association with proapoptotic

factors, activates a family of proteolytic enzymes (the caspases),
causing cleavage of key proteins and resulting in the morphological
and biochemical changes characteristic of apoptosis.
SHARI R BABU
GLYCOGEN METABOLISM
SHARI R. BABU
SCHOOL OF MEDICINE
shari.babu@unza.zm
BIOMEDICAL IMPORTANCE
• Glycogen is the major storage form of carbohydrate in animals, it is a
branched polymer of α-D-glucose.
• It occurs mainly in liver and muscle.
• Small amounts are found in brain, SB1

heart, kidneys, adipose tissues
and erythrocytes.
• Because of its greater mass muscle contains about three to four times
as much glycogen as the liver.
• Muscle glycogen - source of glucose for glycolysis during muscle

contraction.
• Liver glycogen - maintain blood glucose during periods of fasting.

SHARI BABU
Slide 2
SB1 Recent studies show that in the hippocampus, glycogen plays an important role in memory formation and learning.
• Glycogen is present in the cytosol as granules which contain both
regulatory proteins and enzymes that catalyze the synthesis and
degradation of glycogen.
• Liver glycogen stores increase during the well-fed state and are
depleted during a fast.
• Muscle glycogen is synthesized to replenish muscle stores after they

have been depleted following strenuous exercise.
• Glycogen metabolism occurs in the cytosol of liver and muscle cells.
SHARI BABU
Why Store Glucose as Glycogen?
• Three reasons why glycogen is a good fuel reserve

compared to fat are:
a) Fatty acids cannot be released from fat as rapidly as

glucose can be released from glycogen.
b) Fat cannot be used as a source of energy in the absence of

oxygen.
c) Fat cannot be converted to glucose, which is required by

the brain and erythrocytes.
SHARI BABU
GLYCOGEN BIOSYNTHESIS (GLYCOGENESIS)
• Glucose is phosphorylated to glucose 6-phosphate, catalyzed by
hexokinase in muscle and glucokinase in liver.
• Glucose 6-phosphate is isomerized to glucose 1-phosphate by

phosphoglucomutase.
• Next, glucose 1-phosphate reacts with UTP to form UDPGlc which is

the direct glucose donor, catalyzed by UDPGlc pyrophosphorylase.
• An enzyme, glycogenin, acts as a primer to initiate glycogenesis.

• Glycogenin initiates glycogen synthesis by autoglycosylation, transferring
glucose from UDPGlc to itself.
• Once 7-10 glucose residues are added, glycogen synthase takes over.
SHARI BABU
Glucose
Hexokinase/Glucokinase
Glucose-6-phosphate
Phosphoglucomutase
Glucose-1-phosphate + UTP
UDPGlc pyrophosphorylase
UDP-Glucose
Glycogenin
Autoglycosylation UDP-Glc
SHARI BABU
Short chain of glycogen
• Glycogen synthase catalyzes
• the transfer of glycosyl unit from UDP-glucose to the growing glycogen
chain, and
• formation of a α (1→4) glycosidic bond between C1 of the activated
glucose of UDPGlc and C4 of a terminal glucose residue of glycogen,
liberating UDP.
• Glucose is always added to the non-reducing end of the glycogen

chain.
• The glycogen branching enzyme (1, 4-α glucan branching

enzyme) introduces branch points by forming α (1→6) glycosidic
bonds at regular intervals.
• The branching enzyme transfers glucose units from the outer ends of the
glycogen chain to form the α (1→6) glycosidic bond.
SHARI BABU
N.V. Bhagavan, Chung-Eun Ha, in Essentials of Medical SHARI BABU
Biochemistry (Second Edition), 2015
• The branches grow by further additions of 1→4-glycosyl units and
further branching.
• Glycogen has branches located, on average, every 10-14 glycosyl

residues apart, resulting in a highly branched, tree-like structure that
is highly soluble.
• Branching also increases

• the number of non-reducing ends to which new glycosyl residues
can be added and, from which these residues can be removed, and
• the solubility and size of the molecule.
SHARI BABU
GLYCOGEN DEGRADATION (GLYCOGENOLYSIS)
• Glycogenolysis takes place in the cytosol as well as in the

lysosomes.
• Glycogen phosphorylase catalyzes the rate-limiting step in

glycogenolysis by promoting the phosphorylytic cleavage of the α
1→4 bonds to yield glucose 1-phosphate.
• The terminal glucose residues from the non-reducing ends of the

glycogen molecule are removed sequentially until approximately four
glucose residues remain on either side of a α 1→6 branch point.
• Glucan transferase then transfers a trisaccharide unit from one

branch to another, exposing the α 1→6 branch point.
SHARI BABU
• Hydrolysis of the α 1→6 linkages requires the debranching enzyme
(amylo-a (16)-glucosidase) which releases one glucose molecule, and
further phosphorylase action can proceed.
• The combined action of phosphorylase, glucan transferase and

debranching enzyme leads to complete breakdown of glycogen.
Biochemistry. 5th edition. SHARI BABU

Berg JM, Tymoczko JL, Stryer L.
New York: W H Freeman; 2002.
• Glucose 6-phosphate can be formed from glucose 1-phosphate by the
action of phosphoglucomutase.
• In the liver (but not in muscle), glucose 6-phosphatase

hydrolyzes glucose 6- phosphate to glucose that is exported leading
to increased blood glucose concentration.
• Glucose 6-phosphate is retained by muscle, which needs a large

amount of fuel for the generation of ATP.
• Lysosomal degradation of glycogen

• A small amount (1–3%) of glycogen is continuously degraded by
the lysosomal enzyme, acid a-glucosidase [a(14)-glucosidase or
acid maltase].
SHARI BABU
REGULATION OF GLYCOGENESIS &
GLYCOGENOLYSIS
• Cyclic AMP (cAMP) integrates the regulation of

glycogenolysis and glycogenesis.
• The principal enzymes controlling glycogen metabolism— glycogen

phosphorylase and glycogen synthase—
• are regulated by reversible phosphorylation and
dephosphorylation of enzyme protein in response to hormone
action.
• Cyclic AMP (cAMP) is formed from ATP by adenylyl cyclase and

acts in response to hormones such as epinephrine and glucagon.
SHARI BABU
• Increased cAMP leads to phosphorylation of proteins resulting in
their activation or inhibition.
• Glycogen synthase exists in either a phosphorylated (inactive) or

dephosphorylated state (active).
• Phosphorylation of glycogen phosphorylase activates it and

dephosphorylation inhibits it.
• Insulin promotes glycogenesis by activating glycogen synthase via

the action of protein phosphatase-1.
• Glycogen phosphorylase is also allosterically inhibited by glucose 6-

phosphate, as well as by ATP, a high-energy signal in the cell.
SHARI BABU
SB3
SB6
SHARI BABU
Lippincott’s Illustrated Reviews: Biochemistry, Sixth
Edition (2014)
Slide 16
SB3 When blood glucose levels are low, glucagon and/or epinephrine are released. These hormones bind to their respective receptors. The hormone
receptor complex activates the enzyme adenylyl cyclase which converts ATP to cAMP.
High levels of cAMP will activate the enzyme cAMP-dependent protein kinase (PKA).
PKA in the inactive form exist as a tetramer, with two catalytic domain and two regulatory domain. When cAMP levels are high, it will bind to the
regulatory subunits which frees and activates the catalytic subunit. Now PKA is activated to carry out its catalytic function.
The protein kinase will phosphorylate and inactivate glycogen synthase, causing the inhibition of glycogenesis.
Increased secretion of insulin indicates high levels of blood glucose which is favourable for glycogenesis. Insulin activates phosphoprotein
phosphatase enzyme which brings about the dephosphorylation and activation of glycogen synthase.
SB6 Shari Babu, 29/03/2021

SB4
SHARI BABU
Lippincott’s Illustrated Reviews: Biochemistry, Sixth
Edition (2014)
Slide 17
SB4 When glucagon and/or epinephrine are released. These hormones bind to their respective receptors. The hormone receptor complex activates the
enzyme adenylyl cyclase which generates cAMP.
High levels of cAMP will activate the enzyme cAMP-dependent protein kinase.
The protein kinase will phosphorylate and activate glycogen phosphorylase kinase, causing the phosphorylation and activation of glycogen
phosphorylase.
Ca2+ can bind to phosphorylase kinase enzyme and activate it without phosphorylation, which then activates glycogen phosphorylase.
The above mechanisms brings about glycogen degradation.
Insulin will activate protein phosphatase-1 which will dephosphorylate and inactivate phosphorylase kinase, which in turn will stop the activation of
glycogen phosphorylase. Protein phosphatase-1 can bring about the direct dephosphorylation and inactivation of glycogen phosphorylase. This will
halt glycogenolysis.
In liver
• Glucagon leads to formation of cAMP and activation of glycogen
phosphorylase.
In the muscle
• Muscle lacks both glucagon receptors and glucose 6-phosphatase.
• Muscle glycogenolysis is activated;

• In response to epinephrine through β-receptors
• This occurs during
• Flight and fight situation
• Prolonged physical exercise
• Hormone independent mechanisms

• The stimulatory effects of Ca2+ and AMP ensure that the muscle can
respond to its energy needs, even in the absence of hormonal input.
SHARI BABU
PATHWAY GLYCOGENESIS GLYCOGENOLYSIS
REG. ENZYME Glycogen synthase Glycogen phosphorylase
ACTIVE/ACTIVATOR Glycogen synthase a Glycogen phosphorylase a

(dephosphorylated) (phosphorylated)
Insulin  Protein phosphatase-1 Glucagon/Epinephrine 

cAMP  PKA  glycogen
phosphorylase kinase
INACTIVE/INHIBITOR Glycogen synthase b Glycogen phosphorylase b

(phosphorylated) (dephosphorylated)
Glucagon/Epinephrine  cAMP Insulin  Protein

 PKA phosphatase-1
Glycogen Storage Diseases
• A series of inborn errors of metabolism, the glycogen storage
diseases, result from deficiencies in the enzymes of glycogen
metabolism.
• Muscle glycogen phosphorylase is genetically different from liver

glycogen phosphorylase.
• Some degradation of glycogen also occurs within lysosomes when

glycogen particles become surrounded by membranes that then fuse
with the lysosomal membranes.
• A lysosomal glucosidase hydrolyzes this glycogen to glucose.
SHARI BABU
• The enzymatic defect in von Gierke disease (type I) is glucose 6-
phosphatase.
• Glycogen accumulation in the liver.
• The absence of glucose 6-phosphatase SB5

in the liver causes hypoglycemia
because glucose cannot be formed from glucose 6- phosphate.
• The presence of excess glucose 6-phosphate triggers an increase in

glycolysis in the liver, leading to a high level of lactate and pyruvate in
the blood.
• A genetic defect of lysosomal glucosidase, called Pompe disease

(type II glycogen storage disease), leads to the accumulation of
glycogen particles in large, membrane-enclosed residual bodies.
• The accumulation of lysosomal glycogen affects various body tissues,

particularly heart, skeletal muscle, liver and nervous tissues.
SHARI BABU
Slide 21
SB5 Glucose-6-phosphatase deficiency can cause severe hypoglycemia since free glucose is not released from glycogenolysis and glucose is not produced
via gluconeogenesis.
• In Cori (type III) disease, the structure of liver and muscle
glycogen is abnormal, and it accumulates impairing the function of
the tissues.
• Patients having this type lack the debranching enzyme (α-1,6-

glucosidase), and so only the outermost branches of glycogen can be
effectively utilized.
• Thus, only a small fraction of this abnormal glycogen is functionally

active as an accessible store of glucose.
• An individual with McArdle disease (type V glycogen storage

disease) experiences no other symptoms but unusual fatigue and
muscle cramps during exercise.
• Muscle glycogen phosphorylase is the affected enzyme in this

disorder.
SHARI BABU
• Liver glycogen is the first reservoir for the support of blood glucose
levels.
• A deficiency in liver glycogen phosphorylase in Hers or (type

VI) glycogen storage disease) can result in fasting hypoglycemia.
• The hypoglycemia is usually mild because patients can still

synthesize glucose from gluconeogenesis.
• Accumulation of glycogen can lead to liver enlargement.
SHARI BABU
SCHOOL OF MEDICINE
AMINO ACID METABOLISM

Conference Room, Adult Extension Studies Building,
Ridgeway Campus
1
OUTLINE
 Objective of this topic:
• Outline the synthesis (anabolism) and breakdown

(catabolism) of amino acids in the body.
 As we discuss this:
 Clinically important amino acids
 Metabolic defects
2
Dr. Mwale, 2023
OVERVIEW of AMINO ACID METABOLISM
CATABOLISM
1.
Removal of the amino
group and processed to
produce urea
2.
Processing of the
3. remaining carbon
-Transamination reactions skeleton into products
-Amidation reactions
-Synthesis from other amino acids

Ketogenic and glucogenic
products
3
Dr. Mwale, 2023
CATABOLISM-removal of amino group
CATABOLISM

group (oxidative
degradation) and
processed to produce urea
α -keto acids
WHEN/WHY DOES THIS OCCUR?
1. If they are not needed for new protein
synthesis
2. When a diet is rich in protein and the ingested
amino acids exceeds the body’s needs for protein • Undergo oxidation to CO2 and
synthesis, the surplus is catabolized BECAUSE amino H2O
acids can not be stored.
• More frequently provide three-
3. During starvation or in diabetes mellitus when
or four-carbon units that can
carbohydrates are either unavailable or not
be converted by
properly utilized, cellular proteins are used as
gluconeogenesis into glucose
fuel.
4
Dr. Mwale, 2023
CATABOLISM-removal of amino group cont…
• Most amino acids are metabolized in the liver
• Ammonia is generated in the process
• Some of the ammonia is recycled and used in a variety of biosynthetic process
• Excess ammonia is excreted
– Directly as ammonia(In ammonotelic animals as most aquatic vertebrates
such as bony fish)
– After being converted to urea (In ureotelic animals such as many
terrestrial vertebrates)
– After being converted to uric acid (in uricotelic animals such as birds
and reptiles)
NOTE:
– Ammonia is toxic to animal tissues
– Symptoms of ammonia intoxication include a flapping tremor, slurring of speech ,
blurring of vision and in severe cases coma and death
– SO the excess ammonia is converted to a nontoxic compound before export from
extrahepatic tissues to the liver and kidneys via the blood stream
Dr. Mwale, 2023 5

METABOLIC FATES OF AMINO GROUPS
Cellular protein Four Stages of Urea
1 LIVER Biosynthesis.
Amino 1. pyruvate
acids Transaminase/ − 2. acetyl CoA,
COO − COO
from + aminotransferase 3. acetoacetyl CoAI. Transamination
ingested H3N C H C O 4. α-ketoglutarate,
5. Succinyl CoA, II. Oxidative
protein PLP
R
Amino acids transamination
R deamination of
α-keto acids 6. Fumarate glutamate
I 7. oxaloacetate
COO− COO−
+ III. Ammonia transport
C O H3N C H IV. Reactions of the urea
CH2 CH2 Glutamate cycle
α-ketoglutarate
CH2 CH2
3 COO− COO−
Alanine COO− 2
from +
H3N C H Glutamine from
muscle COO− muscle and
+ COO− NH4+ II CH2 other tissues
H3N C H
C O CH2
CH3 Glutamine
Alanine CH3 C
Pyruvate Ammonia, NH 2
urea , uric acid III &IV O 30 6
Dr. Mwale, 2023
STAGE 1: TRANSAMINATION NOTE:
1.There are a number of
Cellular protein
1 LIVER different aminotransferases
Amino 1. pyruvate which are named for the amino
acids Transaminase/ 2. acetyl CoA, group donor eg alanine
COO− aminotransferase COO−
from + 3. acetoacetyl CoA aminotransferase (ALT) also
ingested H3N C H C O 4. α-ketoglutarate, called glutamate-pyruvate
protein PLP transaminase, (GPT) , aspartate
R 5. Succinyl CoA,
R
6. Fumarate aminotransferase (AST) also
α-keto acids called glutamate-oxaloacetate
I 7. oxaloacetate
− −
transaminase, (GOT),
COO COO
+
C O H3N C H 2. Measurements of GPT and
CH2 Glutamate GOT in blood serum is
CH2
α-ketoglutarate important in medical diagnosis
CH2 CH2 of heart and liver damage
caused by heart attack, drug
3 COO− COO− toxicity or infection.
Alanine COO− 2
from
4. After a heart attack SGPT, SGOT (S for serum) and serum creatine + kinase can provide
Glutamine from
muscle
information about severity of the damage. H3N C H 3. Various enzymes may leak
COO− muscle and
due to liver damage but
• Creatine
+ kinase is COO
−
the first enzyme
NH4 + to II
appear after a heart
CH attack;
2 it other
also tissues
aminotransferases are most
H3N C quickly
disappears H from the plasma
C O and GPT follows later CH2 useful for monitoring because
• GOT is the next to appear they are very active in the
CH3 Glutamine
• Lactate dehydrogenase
CH3 also leaks from injured heart muscle liver
Alanine C
• SGOT and SGPTPyruvate
tests are important to determine whether people
Ammonia, NH exposed to
carbon tetrachloride, chloroform etc have
urea III &IV
, uricsuffered
acid
O
liver damage.
2 30 7
Dr. Mwale, 2023
STAGE 2: OXIDATIVE DEAMINATION
• In hepatocytes, Cellular
glutamate
protein is
1
transported from the cytosol into LIVER
Amino 1. pyruvate
the mitochondria where it
Transaminase/
acids − COO− 2. acetyl CoA,
undergoes oxidativeCOO
from deamination
aminotransferase 3. acetoacetyl CoA
+
catalyzed
ingested H3by
N C L-glutamate
H C O 4. α-ketoglutarate,
dehydrogenase
protein PLP
5. Succinyl CoA,
R R
• The enzyme Amino
can useacids
either NAD + or
transamination
α-keto acids 6. Fumarate
NADP+ as acceptor of reducing 7. oxaloacetate
equivalents COO−
I COO−
+
• The combined action
C O of an H3N C H
aminotranferase and glutamate
CH2 Glutamate
dehydrogenase is CH
referred
2 to as L-glutamate is the only amino
α-ketoglutarate
transdeamination CH2 CH2 acid that can undergo
The α-ketoglutarate oxidative deamination at an
• 3 COOproduced
− by
COO− appreciable rate
deamination of glutamate can be
Alanine
used in the citric acid cycle and for +
COO −
2
from Glutamine from
gluconeogenesis
muscle COO−
H3N C H
muscle and
H3N C H
C O CH2
CH3 Glutamine
Dr. Mwale,Alanine
2023 CH3 C
STAGE 3: AMMONIA TRANSPORTATION VIA GLUTAMINE
-Excess ammonia generated in most otherprotein
Cellular tissues
is converted to the 1 amide nitrogen of glutamine LIVER
Amino 1. pyruvate
(labeled as 2), acids
which passes to the liver, thenTransaminase/
into 2. acetyl CoA,
the mitochondria of hepatocytes. + COO
−
aminotransferase COO−
from 3. acetoacetyl CoA
-L-glutamine plays a role in transport
ingested H3N C of H ammonia C O 4. α-ketoglutarate,
from extrahepatic tissues
protein PLP
R 5. Succinyl CoA,
R
HOW?Amino acids transamination
First glutamate + ATP react to form ADP and a γ- I 7. oxaloacetate
− COO−
glutamyl phosphate intermediateCOO that reacts
+
with ammonium to produce glutamine
C O and H3N C H
inorganic phosphate (glutamine synthetase) CH2 Glutamate
CH2
• Glutamine in excess of that required for α-ketoglutarate
biosynthesis is transported in the blood
CH2 to the CH2
liver and kidneys for processing
3 COO− COO−
– The enzyme glutaminase converts glutamine
Alanine
to glutamate
COO− 2
from and NH . Provides another
+4 +
Glutamine from
source of ammonia to be disposed
muscle of by H3N C H
COO − muscle,
urea synthesis COO− II
+ NH4+ CH2 extrahepatic and
– Glutamate is further
H3Nprocessed
C H in the liver other tissues
by glutamate dehydrogenase, releasing
C more
O CH2 transports ammonia
CH3 skeletons for
ammonia and producing carbon Glutamine
CH3 and so serves as a
metabolic fuel Alanine Ammonia,
C
NH 2 non-toxic transport
Pyruvate III &IV O 30 9
Dr. Mwale, 2023 urea , uric acid form of ammonia
STAGE 3: AMMONIA TRANSPORT VIA ALANINE
Cellular protein GLUCOSE- ALANINE CYCLE
1 LIVER
Amino 1. pyruvate
COO − COO
from + aminotransferase 3. acetoacetyl CoA
protein PLP
R 5. Succinyl CoA,
R
I 7. oxaloacetate
COO− COO−
+
C O H3N C H
CH2 CH2 Glutamate
α-ketoglutarate
CH2 CH2
3 COO− COO−
Alanine COO− 2
from +
muscle COO− muscle and
H3N C H
C O CH2
CH3 Glutamine
Alanine CH3 C
Dr. Mwale, 2023
STAGE 3: AMMONIA TRANSPORT VIA ALANINE
Cellular protein GLUCOSE- ALANINE CYCLE
1 LIVER
• Amino
In muscle amino groups are collected in the
Transaminase/
1. pyruvate
acids
form of glutamate by transamination
COO − COO− 2. acetyl CoA,
Glutamate canH3N C H
• ingested C O 4. α-ketoglutarate,
– Be converted to glutamine forPLP
protein
R transport to 5. Succinyl CoA,
R
the liverAmino acids transamination
– Transfer its amino group toI pyruvate 7. oxaloacetate
−
forming alanineCOOcatalyzed by alanine amino COO−
+
transferase C O H3N C H
• Alanine passes
CH2 into the blood stream and CH2 Glutamate
travels to the liver
α-ketoglutarate
CH2
• In the CHcytosol
2 alanine amino
3 transferase transfers
COO − the amino group COO−
Alanine from alanine to α-ketoglutarate COO− 2
from forming pyruvate and glutamate +
muscle muscle and
• Glutmate
COO− then enters the mitochondria
+
where the
COO −
glutamate dehydrogenase
NH4+ II CH2 other tissues
H3N C H
reaction releases C ONH4+ or can undergo CH2
transamination
CH 3 reaction with Glutamine
Alanine CH
oxaloacetate to3form aspartate, another C
nitrogen donor in the urea cycleurea , uric acid III &IV O 30 11
Dr. Mwale, 2023
Amino acids STAGE 4: THE UREA CYCLE
(EXCEPT lysine, threonine, proline
and hydroxyproline) Glutamine
Transamination to α-
from
ketoglutarate
extrahepatic
Glutamate tissues Argininosuccinate synthetase
Alanine
(from muscle) 2a
ATP PPi
Cytosol Citrulline Aspartate
transcarbamoylase
2b Argininosuccinate
synthetase
AMP
Glutamine
Ornithine
Glutaminase 2ATP 2ADP + Pi Citrulline
HCO3− Argininosuccinate
Pi
Oxaloacetate Glutamate NH4 +
Aspartate amino Gluta Carbamoyl 1 3 Argininosuccinate lyase
transferase mate phosphate
Fumarate
Aspartate α-ketoglu. dehyd
rogen Ornithine
ase Mitochondrial Arginine
matrix
H2O
Ornithine Arginase
4
To step 2b of the Urea
urea cycle O
NH2 C NH2 12
Dr. Mwale, 2023
Nitrogen excretion and the urea cycle
• In ureotelic organisms, the ammonia deposited in the mitochondria of the hepatocytes is
converted to urea in the urea cycle
• Urea production occurs mainly in the liver and is the fate of most of the ammonia
channeled there
• Urea passes into the blood stream to the kidneys where its excreted
Urea is produced from ammonia in five enzymatic steps
• Two main SOURCES of the first amino group to enter the cycle
– Ammonia from glutamate and glutamine
– From the intestine via the portal vein where its produced by bacterial oxidation of
amino acids
• Ammonia together with CO2 (as HCO3−) produced by the mitochondrial respiration form
carbamoyl phosphate in the mitochondrial matrix
– ATP- dependent reaction catalyzed by carbamoyl phosphate synthetase I, a regulatory
enzyme
• Carbamoyl phosphate ( an activated carbamoyl group donor) enters the cycle which has four
enzymatic steps
– Carbamoyl phosphate donates its carbamoyl group to ornithine to form citrulline, with
release of Pi (Step 1)
– Catalyzed by ornithine transcarbamoylase. Citrulline that results passes
41 from
13 the
mitochondria into the cytosol Dr. Mwale, 2023
• The second amino group is introduced by aspartate by a condensation reaction
between the amino group of aspartate and the ureido group of citrulline, forming
argininosuccinate (Step 2)
– Reaction is catalyzed by argininosuccinate synthetase. Requires ATP and passes
through a citrullyl-ATP intermediate
• The argininosuccinate is then reversibly cleaved by argininosuccinate lyase to form
free arginine and fumarate (Step 3)
– Fumarate enters the mitochondria to join a pool of citric acid cycle intermediates
• Arginase then splits the arginine to yield UREA and ornithine (Step 4)
– Ornithine is transported into the mitochondria to initiate another round of the
urea cycle
• Aspartate formed in the mitochondria by transamination between oxaloacetate and
glutamate can be transported into the cytosol, where it serves as nitrogen donor in
the urea cycle reaction catalyzed by argininosuccinate synthetase
14
Dr. Mwale, 2023
REGULATION OF CYCLE AT TWO LEVELS
• The flux of nitrogen through the urea cycle varies with the organism's diet
– On high protein diet , the carbon skeletons of amino acids are used for fuel
producing much urea from excess amino groups
– During prolonged starvation, breakdown of muscle protein supplies much of the
metabolic energy and this results in increased urea production
• Allosteric regulation of carbamoyl phosphate synthetase I is activated by N-
acetylglutamate
Pathway interconnections reduce the energetic cost of urea synthesis

• The synthesis of one molecule of urea requires four high-energy phosphate groups
– Two are required to make carbamoyl phosphate
– A second required to make argininosuccinate undergoes the pyrophosphate cleavage
to AMP and PPi which is hydrolyzed to 2Pi
– Urea cycle also causes a net conversion of oxaloacetate to fumarate (via aspartate),
and the regeneration of oxaloacetate produces NADH in the malate dehydrogenase
reaction.
– Each NADH can generate up to 2.5 ATPs reducing the overall energetic cost urea
synthesis
15
Dr. Mwale, 2023
The urea cycle and the citric acid cycle can be linked via
fumarate – aspartate-argininosuccinate shunt
Fumarate
Arginine
Malate
NAD+ Urea cycle
NADH
Oxaloacetate
Aspartate Argininosuccinate Ornithine
argininosuccinate
shunt Aspartate
Citrulline
Aspartate Citrulline
Ornithine
α-ketoglutarate
Oxaloacetate
NADH Glutamate
NAD+
Malate CAC
Fumarate
Dr. Mwale, 2023

METABOLIC FATES OF AMINO GROUPS
Cellular protein
Amino
LIVER 1. pyruvate
COO − COO
protein PLP
R 5. Succinyl CoA,
R
7. oxaloacetate
COO− COO−
+
C O H3N C H
CH2 CH2 Glutamate
α-ketoglutarate 1
CH2 CH2
3 COO− COO−
Alanine COO− 2
from +
muscle H3N C H Glutamine from
COO−
+ COO− CH2 muscle and other
NH4+ tissues
H3N C H
C O CH2
CH3 Glutamine
Alanine CH3 C
O 30 17
Dr. Mwale, 2023 urea , uric acid
Catabolic Fates of the amino acid carbon skeletons.
Tryptophan 1. Pyruvate
Glucogenic Ketogenic
2. Acetyl CoA
Alanine 3. Acetoacetyl CoA
4. α-ketoglutarate
1 5. Succinyl CoA,
6. Fumarate
7. Oxaloacetate
Glucogenic
2 3
Other transamination rxn:

Asp to Oxaloacetate 7
6
Glucogenic
4
Glucogenic 18
Dr. Mwale, 2023
AMINO ACID SYNTHESIS
Introduction:
• Plants and microorganisms can synthesize all
of the 20 standard amino acids.
• Mammals, however, cannot synthesize all 20 and

must obtain some of them in their diet.
• Those amino acids that are supplied in the diet

are referred to as essential, whereas the
remainder that can be synthesized by the
organism are termed non-essential.
1 19
Dr. Mwale, 2023 9
Introduction cont:
2 20
Dr. Mwale, 2023 0
Introduction cont..
• The pathways for the biosynthesis of amino acids are diverse
and often vary from one organism to another.
• But they all have an important feature in common: their
carbon skeletons come from 7 key intermediates in central
metabolic pathways of:
• glycolysis-phosphoglycerate, phosphoenolypyruvate & pyruvate
• citric acid cycle-α-ketoglutarate & oxaloacetate.
• pentose phosphate pathway-ribose 5-phosphate & erythrose 4-

phosphate
Dr. Mwale, 2023 4 21

Introduction cont:
Methionine
2 22
Dr. Mwale, 2023 2
Introduction cont:
2 23
Dr. Mwale, 2023 3
Introduction cont:
Phenylalanine
2 24
Dr. Mwale, 2023 4
INTRODUCTION cont..
• Amino acids are also a source of intermediates and
some biologically essential molecules other than
proteins.
EXAMPLES
• Pyruvate can be formed from glycine, serine, alanine,
cysteine, threonine, 4-hydroxyproline.
• Acetyl CoA can be formed from tyrosine,

phenylalanine, lysine and tryptophan.
• LOOK AT CITRIC ACID CYCLE AND

GLUCONEOGENESIS FOR OTHER EXAMPLES OF
AMINO ACIDS BEING CONVERTED TO METABOLIC
INTERMEDIATES.
25
Dr. Mwale, 2023
• Some OTHER molecules other than proteins
derived from amino acids include:
1. porphyrins where glycine is a precursor
2. synthesis of phosphocreatine requires glycine,

arginine and methionine
3. synthesis of glutathione requires glutamate,

cysteine and glycine.
• These are shown in the next 3 slides:

26
Dr. Mwale, 2023
1. Glycine as a precursor of porphyrins
Succinyl-CoA+ Glycine α-amino-β-ketoadipic acid
δ-aminolevulinic acid
Porphobilinogen (formed
from two molecules of δ-
aminolevulinic acid)
Protoporphyrin IX (formed from 4 molecules of porphobilinogen
27
Dr. Mwale, 2023
2. Creatine is synthesized from glycine and arginine with
methionine in the form of S-adenosylmethionine as the
methyl group donor
Glycine
Arginine
Amidinotransferase
Ornithine
Guanidinoacetate
S-adenosyl methionine
Methyltransferase
S-adenosylhomocysteine
Creatine
Creatine ATP
kinase ADP
Phosphocreatine
28
Dr. Mwale, 2023
3. Biosynthesis of glutathione
• Glutathione is derived from glycine, glutamate and cysteine
• The γ-carboxyl group of glutamate is activated by ATP to form an acyl
phosphate intermediate, which is then attacked by the α –amino group of
cysteine
• A second condensation reaction follows with the α –carboxyl group of cysteine
activated to an acyl phosphate to permit reaction with glycine
Cysteine ATP ADP + Pi Glycine ATP ADP + Pi
Glutamate γ-Glu−Cys γ-Glu−Cys−Gly

γ-Glutamyl Glutathione
synthetase Glutathione
cysteine synthetase
• Glutathione helps maintain the sulfhydryl groups of proteins in the reduced

state and the iron of heme in the ferrous state (see PPP)
• Its redox function is also used to remove toxic peroxides
29
Dr. Mwale, 2023
Catabolic Fates of the Amino Acid Carbon Skeletons.
Tryptophan
Glucogenic Ketogenic
Alanine
Glucogenic
2 3
6
Glucogenic
4
Glucogenic 30
Dr. Mwale, 2023
From 2 & 3: Catabolic pathways for tryptophan, lysine, phenylalanine,
tyrosine, leucine, and isoleucine
Tryptophan Lysine Phenylalanine
9 steps 4 steps Leucine
Tyrosine
Alanine
α-Keto adipate 5 steps
6 steps
CoA-SH NAD+
Fumarate
Pyruvate Acetyl-CoA
CO2 NADH Acetoacetate
Acetyl-CoA Glutaryl-CoA CO2 CoA-SH
4 steps Acetoacetyl-CoA
Isoleucine Acetyl-CoA
6 steps
Propionyl-CoA Succinyl-CoA
3 steps
31
Dr. Mwale, 2023
• Tryptophan yields acetyl-CoA via both pyruvate and acetoacetyl-CoA
– Some intermediates of tryptophan catabolism are precursors of biomolecules
such as nicotinic acid (a precursor of NAD and NADP in animals) and
serotonin (a neurotransmitter in vertebrates)
Phenylalanine catabolism is genetically defective in some people
• A genetic defect in phenylalanine hydroxylase the first enzyme in the catabolic
pathway of phenylalanine is responsible for the disease phenylketonuria (PKU)
• Phenylalanine hydroxylase
– is a mixed –function oxidase
– Requires the cofactor tetrahydrobiopterin which carries electrons from
NADH to O2 and becomes oxidized to dihydrobiopterin
– The cofactor is reduced by dihydrobiopterin reductase
NAD+ Phenylalanine
5,6,7,8-
tetrahydrobiopterin O2
Dihydrobiopterin
(THP)
Phenylalanine hydroxylase
reductase
H2O
NADH + H+ 7,8-Dihydrobiopterin Tyrosine

32
Dr. Mwale, 2023
Catabolic pathways for phenylalanine and tyrosine and associated diseases
THP
Χ
NADH + H+ NAD+
Phenylalanine Tyrosine
Hydroxyphenylpyruvate
Phenylalanine
hydroxylase
Χ Phenylketonuria Χ
Maleyacetoacetate Homogentisate
Maleyacetoacetate
isomerase
Χ
Fumarylacetoacetate
H2O
Fumarylacetoacetase
Χ Tyrosinemia I
Fumarate Acetoacetate
Succinyl-CoA
3-ketoacyl-CoA transferase
Succinate
33
Dr. Mwale, 2023 Acetoacetyl-CoA
• In individuals with PKU, a secondary pathway of phenylalanine metabolism
comes into play in which phenylalanine undergoes transamination with pyruvate
to yield phenylpyruvate
– Phenylalanine and phenylpyruvate accumulate in the blood and tissues and are
excreted in the urine
– Much of the phenylpyruvate , rather than being excreted as such is either
decarboxylated to phenylacetate or reduced to phenyllactate
– Phenylacetate gives the urine a characteristic odour
• Accumulation of phenylalanine or its metabolites leads to mental
retardation
Phenylalanine
Pyruvate
PLP Aminotransferase
Alanine
Phenylpyruvate
Phenylacetate Phenyllactate
• Phenylketonuria can also be caused by a defect in the enzyme that catalyzes the
regeneration of tetrahydrobiopterin
Dr. Mwale, 2023 34

From 7: .Asparagine and aspartate are degraded to oxaloacetate
• The carbon skeletons of asparagine and aspartate
ultimately enter the citric acid cycle as oxaloacetate
Asparagine
H2O
Asparaginase
+
NH4
Aspartate
α–ketoglutarate
Aspartate PLP
amonitransferase
Glutamate
Oxaloacetate
• Asparaginase catalyzes the hydrolysis of asparagine to aspartate, which

undergoes transamination with α–ketoglutarate to yield glutamate and
oxaloacetate 25
35
Dr. Mwale, 2023
Metabolic disorders of branched amino acids
(Leucine, valine and isoleucine)
• Branched amino acids are not metabolized in the liver
• They’re metabolized in extrahepatic tissues with an aminotransferase not

found in the liver
• The aminotransferase acts on all three branched-chain amino acids to

produce corresponding α-ketoacids.
• The branched-chain α-keto acid dehydrogenase complex then catalyzes

the oxidative decarboxylation of all three α-ketoacids, in each case releasing
a carboxyl group as CO2 and producing the acyl-CoA derivative
(Analogous to reactions catalyzed by pyruvate dehydrogenase and α–
ketoglutarate dehydrogenase)..
36
Dr. Mwale, 2023
Metabolic disorders of branched amino acids
(Leucine, valine and isoleucine) cont…
• The three enzyme systems have the same cofactors (NAD, FAD, TPP,
lipoate, CoA).
• In maple syrup urine disease (because of the characteristic odour

imparted to the urine by the α-ketoacids ) the three branched-chain α-
ketoacids accumulate in the blood and spill over in the urine.
• This condition results from defective branched-chain α-keto acid

dehydrogenase complex.
• Untreated, this condition leads to abnormal development of the brain,
mental retardation, and death in early infancy.
• Treatment involves limiting the amounts of valine, leucine and isoleucine to
the minimum required for normal growth.
37
Dr. Mwale, 2023
4.Amino acids are converted to biological amines by decarboxylation
Tyrosine Glutamate Tryptophan

Tetrahydrobiopterin Glutamate
Tetrahydrobiopterin
O2 Tyrosine
H2O hydroxylase
* decarboxylase
PLP
O2 Tryptophan
Dihydrobiopterin CO2 H2O hydroxylase
Dihydrobiopterin
Dopa
PLP Aromatic amino acid γ-Aminobutyrate
decarboxylase
* CO2
(GABA)
5-Hydroxytryptophan
Dopamine Histidine
Ascorbate Histidine PLP
Aromatic amino acid
O2 Dopamine β-
H2O hydroxylase * decarboxylase
PLP * decarboxylase
CO2
Dehydroascorbate CO2
Norepinephrine
Histamine Serotonin
S-adenosylmethionine
S-adenosylhomocysteine
Epinephrine
Phenylethanolamine N-
*=decarboxylation
methyltransferase 38
Dr. Mwale, 2023
FROM THE ABOVE FIGURE;
• Tyrosine gives rise to a family of catecholamines that includes dopamine, norepinephrine,
and epinephrine
– Levels of catecholamines correlate with changes in blood pressure
– Neurological disorder Parkinson’s disease is associated with an underproduction of
dopamine and has been treated by administering L-Dopa
• Glutamate decarboxylation gives rise to γ-aminobutyrate (GABA), an inhibitory
neurotransmitter
– Overproduction of GABA is associated with epileptic seizures
– GABA analogs are used in the treatment of epilepsy and hypertension
• Serotonin, a neurotransmitter is derived from tryptophan
• Histidine is decarboxylated to form histamine, a powerful vasodilator in animals
– Histamine is released in large amounts as part of an allergic response, and also
stimulates acid secretion in the stomach
– Histamine receptor antagonist cimetidine (Tagamet ) is a structural analog of
histamine.
– It promotes healing of duodenal ulcers by inhibiting secretion of gastric acids
14 39
Dr. Mwale, 2023
Arginine is a precursor for biological synthesis of nitric oxide
Arginine Hydroxyarginine Citrulline + NO
• The reaction is catalyzed by nitric oxide synthase which

structurally is related to NADPH cytochrome P-450 reductase
• In humans NO plays a role in a range of physiological processes-

neurotransmission, blood clotting, signal transduction and the
control of blood pressure
Dr. Mwale, 2023 40

OVERVIEW of AMINO ACID METABOLISM
CATABOLISM
1.
group and processed to
produce urea
2.
3.
Processing of the
-Transamination reactions
remaining carbon
-Amidation reactions
skeleton into products
-Synthesis from other amino acids
Catecholamines
GABA -Metabolic Defects
Serotonin Ketogenic and glucogenic
Enzymes involved in transamination
Histidine Urea cycle enzymes products
NO Phenylalanine hydroxylase---PKU
Porphyrins Dihydrobiopterin reductase defect
Creatine----Phosphocreatine Tyrosinemia I, II, III
Glutathione Alkaponuria 41
Medical diagnosis in levels some enzymes Dr. Mwale, 2023
THANK YOU
inprotected.com 42

Biochemistry Notes Test 2 2023

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THE UNIVERSITY OF ZAMBIA

NUTRITION AND FAT SOLUBLE VITAMINS

Mwale Nicholas K.(PhD)

3. Genetics? Find out what this is about

• It can be summarised thus:

• A rough estimate of the BMR may be obtained by assuming it is 24

-This range prevents chronic diseases

How does one know their AMDR?

• They are not essential nutrients, because the carbon

• Absence of dietary carbohydrate leads to ketone body

• The RDA for carbohydrate is set at 130 g/day for

Dr. Mwale, 2023

Dr. Mwale, 2023

• Dietary ω-3 polyunsaturated fats suppress cardiac

• They do not occur naturally in plants, but occur in

• They are formed during the hydrogenation of liquid

Dr. Mwale, 2023

A. 9 ESSENTIAL AMINO ACIDS

-Tyrosine is considered conditionally essential. It is

-Cysteine is also considered as conditionally essential.

Dr. Mwale, 2023

C. PLANT SOURCES OF PROTEIN

1. Positive nitrogen balance

Dr. Mwale, 2023

- Two forms of vitamins: fat-soluble & water-soluble

• Fat-soluble vitamins deficiencies are most likely in patients

• Water-soluble vitamins are transported in blood

Water-Soluble Vitamins Fat-Soluble Vitamins

A) Retinal is the aldehyde derived from the oxidation of

A) Retinal is the aldehyde derived from the oxidation of

• Also needed for normal growth and for mucus

• Also required for the synthesis of the transferrin and

• Vitamin A is also an important antioxidant that may

Release of Vitamin A from the liver

• Retinol is oxidized to retinoic acid which binds with high

• This activated retinoic acid–receptor complex interacts

Acne and psoriasis

• Occasionally, muscle weakness, fatigue, nausea, and

• However, Vitamin E is the least toxic of the fat-soluble

•Note that the carboxylation is sensitive to inhibition by

•There is also an interaction of prothrombin with platelets,

•Animal foods, in general, contain limited amounts of

•Vit.K1 available in green leaves of plants as phylloquinone

WATER SOLUBLE VITAMINS

Mwale Nicholas K.(PhD)

The vitamins of the B complex

• The B vitamins are also important for normal appetite,

• It is also important in transmission of nerve impulses by

• It also helps to release energy from foods as Thiamine

-Chronic peripheral neuritis

-Beriberi (severe muscle weakness and muscle wasting,

• Major symptoms of mild thiamine deficiency include: loss of

• Has a central role in energy yielding metabolism; it

• Flavin coenzymes (FAD and FMN) are the two most

• The main dietary precursors of riboflavin are milk and

• Riboflavin deficiency usually occurs along with other

– Symptoms include cracks at the corners of the

– Riboflavin deficiency can occur in infants receiving

• Niacin strongly inhibits lipolysis in adipose tissue

• Niacin also useful in treating hyperlipoproteinemia

• The liver normally uses these circulating fatty acids as

• So both VLDL and LDL are elevated.

• Roles-Coenzyme for many enzymes in amino acid