03-01-16 L-1

Metabolism Bioenergetics
Carbohydrate disorder
Glycogen
Found in-
1. Liver
2. Muscle- Do not contribute to blood glucose level because glucose 6-phosphatease enzyme is not
found here.
Bioenergetics

Living cells and Organisms must work to stay alive, to grow and to reproduce.
All living organisms have the ability to produce energy and to channel it into biological work.
Living organisms carry out energy transduction. Conversion of one form of energy to another
form.
Bioenergetics is the quantitative study of energy transductions that occur in living cells and of
the nature and function of the chemical process underlying these transductions.
It includes the study of cellular processes such as respiration and many other metabolic processes
that can lead to production and utilization of energy in forms such ad ATP molecule.

The Goals of This lesson

o Review the laws of thermodynamics
o Understand the quantitative relationships among free energy, enthalpy and entropy
o Describe the special role of ATP in biological energy exchanges.
Bioenergetics and Thermodynamics
Biological energy transductions obey the laws of thermodynamics.
1st law
For any physical or chemical changes, the total amount of energy in the universe remain
constant, energy may change from one form or it may be transported from one region to
another but cannot be created or destroyed.
2nd law
The universe always tends toward increasing disorder: in all natural processes the entropy of
the universe increases.

o The reacting system may be an organism, a cell or two reacting compound. The reacting
system and its surroundings together constitute the universe.
o In the laboratory some chemical or physical processes can be carried out in closed systems
and no material or energy is exchanged with the surroundings.
o However living organism are open systems. They exchange both material and energy with
their surroundings.

04-01-16 L-2
Key pointATP transduction
Living system Open System Never at equilibrium with surrounding Uses free energy Expressed
as G or Gibbs free energy
Amount of energy capable of doing work during a reaction at constant temperature and pressure.
If G (-) reaction is exergonic
If G (+) reaction is endergonic
Unit is Joule/mole or calories/mole
Calorie: Energy required to raise the temperature of 1g of water to 1 degree Celsius.
1calorie = 4.186 Joules.
Joule: Energy transferred or work done when applying force of 1N through a distance of 1m.
G = H TS, (under constant temperature and pressure)
Change in free energy is related to enthalpy and entropy
Enthalpy: Heat content of reacting system. H reflects the number of kind of chemical bonds in the
reactants and products.
When a chemical reaction releases energy it is said to be exothermic.
Heat content of product < Heat content of reactant.
When absorbs energy it is endothermic.
Heat content of product > Heat content of reactant
Unit joules/mole or calories/mole
Entropy: Quantitative expression for the randomness and disorder of the system.
Joules/mole. Kelvin
How Biological System maintain Order?
Living organisms preserve their internal order by taking free energy from their surroundings in the form
of nutrients or sunlight and returning to their surroundings an equal amount of energy as heat and
entropy.
Example: The oxidation of glucose
Aerobic organisms extract free energy from glucose obtained from their surrouindings by oxidizing the
glucose with oxygen (also obtained from surroundings). The end products of this oxidation reaction are
CO2 and H2O and they are returned to the surroundings. At the end of this process, the surroundings
undergo an increase in entropy, whereas the organism itself remains in a steady state and no change
occurs in its internal order.

Whenever a chemical reaction results in an increase in the number of molecules the entropy of the
surroundings increases.
Whenever a solid substance is converted into liquid or gaseous forms the entropy of the surroundings
increases. Because this transformations allow the molecule more freedom for movement.
*Solid converts liquid/gaseous formEntropy increases
*Because More freedom More entropy
Cells require source of free energy
Acquire free energy from sunlight, nutrient etc. and transform this free energy into ATP or other energy
rich compound.
They are capable of providing energy for biological work at constant temperature.
At Constant temperature and pressure
Standard free energy change is related to equilibrium constant.
The composition of the reacting system changes until equilibrium is reached.
aA + bB cC + dD
At equilibrium, rate of forward reaction = rate of reverse reaction
And no further change occurs in the system.
The Keq is defined by the molar concentrations of products and reactants at equilibrium
[] []
Keq = [] []

24-01-16 L-3
Bioenergetics: The study of the transformation of energy in living organisms.
When a reacting system is not at equilibrium the tendency to move toward the equilibrium
represents a driving force.
The magnitude of this driving force is expressed change in free energy or G.
If temperature is 25C or under standard condition and initial reactant and product concentration is
1M the driving force is defined as standard free energy change G.
But Biological system does not remain at standard condition.
By this definition, Standard state for reactions involves hydrogen ion is [H+] = 1M or pH=0.
However most biological reactions occur in well-buffered aqueous solution near pH=7. Both the
pH and concentration of water (55.5M) are essentially constant.
For convenience of calculations, biochemists define a different standard state in which the
concentration of [H+] is 10-7M and that of water is 55.5M, for reactions that involve Mg2+
(available in most reactions involving ATP), its concentration in solution is commonly taken to be
constant at 1mM.
Physical constants based on this biochemical standard state are called Standard Transformed
Constants and written as G'0 and K'eq to distinguish them from the untransformed constants
which are used by chemists.
 G'0 is the difference between the free energy content of the products and the free energy
contents of the reactants under standard conditions
G'0 = G'0 products G'0 reactives)
When G'0 is negative, the products contain less free energy than the reactants and the reaction
will proceed spontaneously under standard conditions
When G'0 is positive, the products contain more free energy than the reactants and the reaction
will tend to go in the revers direction under standard conditions
Each chemical reaction has a characteristic standard free energy change which may be positive,
negative or zero depending on the equilibrium constant of the reaction
G'0 tell us in which direction and how far a given reaction must go to reach equilibrium when
the initial concentration of each component is 1M, the pH is 7, the temparature is 250C.
Thus G'0 is a constant; a characteristic for a given reaction
Actual free energy change
Actual free energy change (G) is a function of reactant and product concentrations and of the
temparature prevailing during the reaction which will not necessarily match the standard
conditions as defined before.
G of any reaction proceeding spontaneously toward equilibrium is always negative and become
less negative as the reaction proceeds and ultimately becomes zero at equilibrium.
An example:
A+B C+D
Reaction is taking place at the standard temparature and pressure
But the concentrations of A,B,C and D are not equal and none of them at the 1M concentration
In order to determine actual G under these non-standard concentrations as the reaction
proceeds from left to right, we enter the actual concentrations of A,B,C and D in this equation.
Rest of the terms in the equation (R,T, G'0 ) are standard values
When the reaction is at equilibrium there is no force driving the reaction in either direction and
G is zero, thus equation reduces to
[C] [D]
0= G'0 + RTln Standard Transformed Constants
[A] [B]
G' = -RT lnK'eq
0
R=8.315 J/mole.K
If Keq >1 G'0 is negative
If Keq <1 G'0 is positive
If Keq=1 G'0 is Zero.
Phosphogulcomutase

Glucose-1-phosphate Glucose-6-phosphate, @25C and pH=7

[Glucose6phosphate]
Keq= = 19mM/1mM =19
[Glucose1phosphate]
G'0 = -RT lnK'eq = -7.3kJ/mol
Spontaneity of a reaction
The criteria for spontaneity of a reaction is the value of G (free energy change) not G'0
If G'0 is positive, Then G can be negative if RT lnK'eq is more negative than the value of G.
Practically it is done by immediate removal of the product.Then K decreases and RT lnK'eq becomes
negative.

25-01-16 L-4
1000 /
*Molar concentration of water = = 55.5M
18 /
*Reaction should be spontaneous if standard free energy is large negative. But thats always not the case.
*When wood burns its free energy is negative but after some times reaction does not proceed cause
activation energy of CO2 is very high. Enzymes decreases the activation energy in the biological system.
*Bioenergetics deal with how a thermodynamically unfavorable reaction can be turned into favorable
one. This is done by coupling reaction.
The main rule in biochemical reactions in living organisms:
All endergonic reactions are coupled to an exergonic reaction. There is an energy cycle in cells that links
anabolic and catabolic reactions.
This principle of bioenergetics explains how a thermodynamically unfavorable (endergonic) reaction can
be driven in the forward direction by coupling it to a highly exergonic reaction through a common
intermediate.
A B G'01
B C G'02
Sum: A C G'01 + G'02
*Change IN
Free energy additive (G'01 + G'02)
Equilibrium constant multiplication (K1K2)
An example: The first step of glycolysis
Glucose + Pi Glucose 6-phosphate+H2O G'0 =13.8 kj/mol
G'0 >0 reaction is not spontaneous
Another very exergonic cellular reaction: Hydrolysis of ATP
ATP + H2O ADP + Pi G'0 = -30.5 kj/mol
These two reactions share the common intermediates H2O and Pi and may be expressed as sequential
reactions:
Glucose + Pi Glucose 6-phosphate+H2O
ATP + H2O ADP + Pi
Sum: Glucose+ ATP Glucose 6-phosphate+ADP
The overall standard free energy changes: G'0 =13,8 kj/mol + (-30,5 kj/mol)= -16,7 kj/mol
Overall reaction is exergonic.
C16H32O2 Palmitic acid
DigestionAbsorbtionMetaboilismElectrontransoportProduce energy in the form of ATP.
High Energy Compound:
Heat acts as an enzyme in uncatalysed reaction
Fatty acid oxidation, Glucose Oxidation released energy
C6H12O6 + 6O2 6CO2 + 6H2O G0= -2850 kJ/mol
C16H32O6 + 23O2 16CO2 +16H2O G0= -9787kJ/mol
Energy released in every step of the reaction is not used up in the process but conserved in high energy
compound. This stored energy subsequently can be used in endergonic process or high energy
intermediate/ conserved energy energy currency.
Free energy producing reaction paying for those reaction in which energy is consumed.
Why high energy compound/ATP is used as energy currency?
Exergonic reaction releases energy and that energy is consumed in endergonic reaction. Breaking of ATP
or high energy compound releases energy. Thats why they are called energy currency.
Example of high energy compound ATP
Possible source of energy
1. Stored nutrient
2. Ingested Foods
3. Solar energy
Catabolic reaction (exergonic) CO2

Fig: Central role of ATP

 A phosphoanhydride bond is found between two phosphate
groups of the ATP molecule, and this type of bond is what links
two phosphates together, which occurs between the Phosphorus
atom of one phosphate group and the oxygen atom of another
phosphate group. A phosphoester (no diester!!!) bond links
adenosine to a neighboring phosphate. This occurs when the -OH
group of adenosine becomes deprotonated, and it gets attacked
by a phosphate group.

31-01-16 L-5
ATP (Adenosine Tri-Phosphate) =Biological energy currency
NH2
N
N

N N
O O O O
HO P O P O P O OH
O O O OH

Large amount of free energy is released when a phosphodiester bond breaks.

AMP is produced when a pyrophosporyle group is released.
High energy compound IF hydrolysis releases >25kJ/mole energy.
Examples: Phosphoenolpyruvate, G = -61.9 kJ/mole
1, 3 Bisphosphoglycerate, G = -49.4 kJ/mole
Phosphocreatine (Emergency source for energyReserved in muscleReleased when vigorous
exerciseFormed when ATP is available by addition with creatine), G = -43.1 kJ/mole
ATP ADP + Pi, G = -30.5 kJ/mole
ATP AMP + PPi, G= -32.2 kJ/mole
Glucose-6-phosphate, G = -19 kJ/mole
Glucose-3-phosphate, G = -13 kJ/mole
As these two has G<25, so they are not high energy compound.
*Creatine Kinase (CK) is an enzyme of muscle and their measure determines heart function.
FIGURE 131 Chemical basis for the large free-energy change associated with ATP hydrolysis.
1) The charge separation that results from hydrolysis relieves electrostatic repulsion among the four
negative charges on ATP. 2) The product inorganic phosphate (Pi) is stabilized by formation of a resonance
hybrid, in which each of the four phosphorusoxygen bonds have the same degree of double-bond
character and the hydrogen ion is not permanently associated with any one of the oxygen. (Some degree
of resonance stabilization also occurs in phosphates involved in ester or anhydride linkages, but fewer
resonance forms are possible than for Pi.) 3) The product ADP2- immediately ionizes, releasing a proton
into a medium of very low [H] (pH 7). 4) A fourth factor (not shown) that favors ATP hydrolysis is the
greater degree of solvation (hydration) of the products Pi and ADP relative to ATP, which further stabilizes
the products relative to the reactant.
Why ATP is highly exergonic but Kinetically Stable?
*Although the hydrolysis of ATP is highly exergonic (G'0 = -30,5 kj/mol), the ATP is stable at pH 7, because
the activation energy for ATP hydrolysis is relatively high. Rapid hydrolysis of ATP occurs only when
catalyzed by an enzyme.
Activation energy -200 to -400 kJ/mole
Where G0 = -30.5 kJ/mole
Enzymes lowers that activation energy.
*The free energy change for ATP hydrolysis is -30.5 kJ/mole under standard conditions but the actual free
energy change (G) of ATP hydrolysis in living cells is very different.
The cellular concentrations of ATP, ADP and Pi are not same and are much lower than the 1 M standard
conditions.
In addition, Mg2+ in the cytosol binds to ATP and ADP and for most enzymatic reactions that involve ATP
as phosphorly group donor, the true substrate biological system is MgATP-2 (ATP + Mg2+). The relevant
G'0 is therefore that for MgATP-2 hydrolysis.
Gp= G0 + RT ln Keq
Phosphorylation potential (Gp) G (actual free energy change) for biological system.
In erythrocyte, concentration of ADP = .2510-3 , Pi = 1.610-3 , ATP = 2.2510-3
ATP ADP + Pi
Gp = -52kJ/mole (Ranges between -50 to -65 kJ/mole)
01-02-16 L-6
*Actual free energy Phosphorylation potential (Gp)
*Not only hydrolysis but also group transfer can provide energy by ATP

Adenylyl group transfer into the substrate or enzymeRaises free energycovalently bound phosphate
containing moiety then transferred
Non-covalent binding of ATP/GTP followed by hydrolysis of ATP/GTP
o Provide energy which changes conformation of protein
o Muscle contraction
o Movement of DNA polymerase
o Motion of ribosome along mRNA
o G-protein activation
Signaling cascade a series of chemical reactions (Signal transfer) which are initiated by a stimulus (e.g.
Hormone).
*Mixture of all the product will be found when reacted
*Adenylyl moiety transforms thermodynamically unfavorable reactions into favorable.
*Fatty acidFatty acyl CoA, (Fatty acid activation reaction) Adenylyl reaction.
*Fatty acid + CoA + ATP Fatty acyl CoA + AMP +PPi
Next step PPi2Pi
USES OF ATP
1. Assembly of informational macromolecules
Energy is required for the assembly of protein, DNA and RNA from its precursor molecule for both the
condensation of monomeric unit and for the creation of ordered sequence.
*NTP (Nucleoside Tri-phosphate), dNTP (de-oxy Nucleoside Triphosphate) are the monomeric unit of RNA
and DNA. PPi and AMP released by the breakage of bond between and of ATP/GTP. This GMP binds
with the NTP/dNTP.
*In protein synthesis amino acids need to be activated for translation
A.A. + ATP Aminoacyl-AMP + PPi
Aminoacyle-AMP + t-RNA aminoacyl-t-RNA (activated form) +AMP

02-02-16 L-7
2. Active Transport
Energy mediated transport against a concentration gradient.
In resting condition in Human brain and Kidney, 2/3 of total energy consumption used for pumping K+ and
Na+ ion across the membrane via K+ and Na+ ATP synthase.
 3. Muscle contraction
Myosin head and Actin filament.

Myosin head has ATP binding sitewhen ATP bindsDissociates form ActinThen ATP hydrolysis
(ATPADP +Pi) Changes conformation When Pi dissociates Power strokes and goes to next
step/Shifting lastly ADP dissociates and myosin head returns to initial state (attached to actin).
The change in conformation of many individual myosin molecules results in the sliding of myosin fibrils
along actin filaments which translates into macroscopic contraction of the muscle fiber.

Enzyme Nucleoside diphosphate kinase

*Cellular system generally has 10 times more ATP than ADP.
During periods of intense demand for ATP, the cell lowers the ADP concentration, and at the same
time acquires ATP, by the action of adenylate kinase:
 2ADP + Mg2+ ATP + AMP
This reaction is fully reversible, so after the intense demand for ATP ends, the enzyme can recycle
AMP by converting it to ADP which can then be phosphorylated to ATP in mitochondria.
*Phosphocreatine (PCr) serves as a ready source of phosphoryl groups for the quick synthesis of ATP
from ADP. The phosphocreatine concentration in skeletal muscle is considerably higher than those in
the other tissues. The enzyme creatine kinase catalyzes the reversible reaction.
Mg2+ ADP+ PCr ATP+ Cr G'0 = -12.5 kj/mol
*Phosphocreatine concentration is 10 times higher than ATP.
When a sudden demand for energy depletes ATP, the PCr reservoir is used to replenish ATP at a rate
faster than ATP can be synthesized by catabolic pathways.
When the demand for energy slackens ATP produced by catabolism is used to replenish the PCr
reservoir by reversal of the creatine kinase reaction.
Firefly:
LuciferinComplex Carboxylic acid
LucoferaseEnzyme
Luciferyl
Adenylate
Added
ATP-->Pi
Luciferase+Mg2+

O2-->light + CO2 +AMP

Luciferin

Oxyluciferin

Application in molecular biology:

Measure the expression of gene of interest (GOI).
Gene of Luciferase inserted into the plasmid + Luciferine and Mg2+ added into the medium
If plasmid grows properlyexpression vectors expressedgene of interest expresses
Measure the emission of light using Illuminometer.
ATP Cycle:
 ATP
-->Pi Pi-->
ADP
ATPADP + Pi Phosphorylation ATP
Pi can enter in 2 ways
1. Oxidative Phosphorylation (In Mitochondria by ETC)
2. Substrate level Phosphorylation (Pi comes from a substrate)
07-02-16 L-8
General aspects of Metabolism
Metabolism (from Greek: ) metabol, "change" or (Greek: ) metabolisms, "out
throw" is the set of life-sustaining chemical transformations within the cells of living organisms.
These enzyme-catalyzed reactions allow organisms
o to grow and reproduce,
o maintain their structures, and
o Respond to their environments.
The word metabolism can also refer to all chemical reactions that occur in living organisms, including
digestion and the transport of substances into and between different cells, in which case the set of
reactions within the cells is called intermediary metabolism or intermediate metabolism.
Metabolism is usually divided into two categories:
1)Catabolism, that breaks down organic matter and harvests energy by way of cellular respiration, and
2)anabolism that uses energy to construct components of cells such as proteins and nucleic acids.
The chemical reactions of metabolism are organized into metabolic pathways, in which one chemical is
transformed through a series of steps into another chemical, by a sequence of enzymes. The reactants,
intermediates and products are referred to as metabolites.
Enzymes are crucial to metabolism because they allow organisms to drive desirable reactions that require
energy that will not occur by themselves, by coupling them to spontaneous reactions that release energy.
Enzymes act as catalysts that allow the reactions to proceed more rapidly. Enzymes also allow the
regulation of metabolic pathways in response to changes in the cells environment or to signals from other
cells.
The metabolism of a particular organism determines which substances it will find nutritious and which
poisonous.
For example, some prokaryotes use hydrogen sulfide as a nutrient, yet this gas is poisonous to animals.
The speed of metabolism, the metabolic rate, influences how much food an organism will require, and
also affects how it is able to obtain that food.
Basal metabolic rate (BMR)the minimal rate of energy expenditure per unit time (metabolic rate)
by endothermic animals at rest.
Body Mass Index (BMI) is a value derived from the mass and height of an individual. The BMI is defined
as the body mass divided by the square of the body height.
*A striking feature of metabolism is the similarity of the basic metabolic pathways and components
between even vastly different species.
For example, the set of carboxylic acids that are best known as the intermediates in the citric acid cycle
are present in all known organisms, being found in species as diverse as the unicellular bacterium
Escherichia coli and huge multicellular organisms like elephants.
These striking similarities in metabolic pathways are likely due to their early appearance in evolutionary
history, and retained because of their efficacy.
Catabolism (Greek kata = downward + ballein = to throw)
It Is the set of metabolic pathways that breaks down molecules into smaller units to release energy.
In catabolism, large molecules such as polysaccharides, lipids, nucleic acids and proteins are broken down
into smaller units such as monosaccharides, fatty acids, nucleotides, and amino acids, respectively.
The most common set of catabolic reactions in animals can be separated into three main stages.
In the first, large organic molecules such as proteins, polysaccharides or lipids are digested into their
smaller components outside cells.
Next, these smaller molecules are taken up by cells and converted to yet smaller molecules, usually acetyl
coenzyme A (acetyl-CoA), which releases some energy.
Finally, the acetyl group on the CoA is oxidized to water and carbon dioxide in the citric acid cycle and
electron transport chain, releasing the energy that is stored by reducing the coenzyme nicotinamide
adenine dinucleotide (NAD+) into NADH.

Not all amino acids are converted into acetyl CoA. Some of them form oxaloacetate or some pyruvate and
then enters into TCA cycle.
08-02-15 L-9
Experimental approaches to study metabolism:
A metabolic pathway can be understood at several levels:
1) In terms of the sequence of the reactions by which a specific nutrient is converted to end
products, and the energetics of these conversions.
2) In terms of the mechanisms by which intermediate is converted to its successor.
3) In terms of the control mechanisms that regulate the flow of metabolites through the pathway.
This includes the inter organ relationships that adjust metabolic activity to the needs of the entire
organism.
Metabolic studies on intact organisms
In 1921 Frederic Banting and Charles Best performed pancreas experiments on dogs (removed whole
pancreas) to measure sugar in their urine and blood. Over a summer they extracted the first antidiabetic
substance.
Surviving-slice and Manometric methods
Solid animal or plant tissues are sectioned into thin slices in which most of the cells remain intact. The
tissue slices are incubated in a buffered medium with a given metabolite to study its conversion into
metabolic products.
1. Tracing the metabolite fates
2. Perturbing the system
Isotope labeling methods:
A metabolic pathway in which one compound is converted to another can be followed by tracing a
specifically labeled metabolite.
Franz Knoop fed dogs with fatty acids that carried a phenolic group serving for labeling.
He observed that:
When giving the dog an odd-chain fatty acid derivatives, Hippuric acid (glycine derivative with a single
carbon in its acyl chain) was recovered from the dog's urine.
When an even-chain fatty acid derivatives were given, Phenylaceturic acid (glycine derivative with two
carbons in its acyl chain) was left.
Clearly, the acyl chain was not degraded one
carbon at a time, or both types of fatty acids
would have produced hypuric acid.
If degradation proceeded by groups of more than
two carbons, correspondingly larger phenylated
derivatives would be produced.
Franz Knoop therefore proposed that fatty acids
are shortened two carbons at a time by oxidation
at the beta carbon.
Chemical labeling has the disadvantage that the chemical properties of labeled metabolites differ from
those of normal metabolites. This problem is eliminated by labeling molecules with isotopes.
Heme biosynthesis actually start with glycine. This is proved using isotope labeling. If glycine is labeled
with 15N, it is found in heme molecule after the heme biosynthesis. But if Glutamic acid, proline are labeled
with 15N, it is not found in the end product (heme).

10-02-16 L-10
Isotope labeling technique is mostly used to study metabolic pathway.
Experimental approaches to study metabolism:
1. Metabolic studies on intact organisms
The beginning (e.g. how much oxygen is used) and end (e.g how much CO2 and ethanol is produced) of
some major metabolic pathways have been identified from balance sheet studies of the metabolic input
and output of intact organism. This doesnt give idea about the pathway.
2. Surviving-slice and Manometric methods
Solid animal or plant tissues are sectioned into thin slices in which most of the cells remain intact. The
tissue slices are incubated in a buffered medium with a given metabolite to study its conversion into
metabolic products. Monometer is a machine used to measure consumption and excretion of gas. This
doesnt give idea about the pathway.
3. Study in genetic mutants
In genetic mutants, there is defects in one or more enzymes in the biosynthetic pathway. Such defects
can result in the accumulation and excretion of the substrate of the defective enzyme.
E1 E2 E3 E4

A B C D Arg

Suppose there is a defect (mutant) in Enzyme E3. Then Arg will not be produced. But there will be Arg in
wild type. This method gives idea about the pathway.
4. Labeling of metabolite
a. Chemical labeling
b. Isotope labeling.
Glycine is common in Hippuric acid and phenylaceturic acid but their fatty acid part is different.
All 27 carbon atoms of cholesterol are derived from acetyl CoA in a three-stage synthetic process: proved
by using radioactive isotope 14C.

5. Study in cell free systems:very popular

If the cell membrane is disrupted by mild homogenization in isotonic sucrose solution, subcellular
organelles can be isolated by differential centrifugation. Theses fractions can then be tested in vitro for
metabolic studies.

Glycolysis: splits glucose to pyruvate, which can be converted to acetyl CoA or lactate.
Gluconeogenesis: converts pyruvate to glucose.
Glycogenesis: synthesis of glycogen, carbohydrate fuel storage form.
Glycogenolysis: breakdown of glycogen.
Pentose Phosphate Pathway (PPP): produces NADPH for cell biosynthesis.
Citric Acid Cycle: converts Acetyl CoA to CO2 and energy
Hypoglycemiclow blood sugar
Carbohydrate metabolism
Plants storage carbohydrate Starch
Animal storage carbohydrate glycogen
Amylase 1. Pancreatic 2. Salivary
15-02-16 L-11
Carbohydrate digestion
o In the mouth, the salivary enzyme amylase begins to hydrolyze starch into short
polysaccharides and maltose.
o In the stomach, acid continues to hydrolyze starch while fiber delays gastric emptying and
provides a feeling of fullness (satiety).
o In the small intestine, pancreatic amylase among other enzymes (maltase, sucrase, and
lactase) hydrolyzes starches to disaccharides and monosaccharides.
o In the large intestine, fibers remain and attract water, soften stools and ferment.
Limit dextrin fragments of amylopectin containing (1-6) branch points. Enzyme Dextrinase
Fiber: 1) Soluble (i.e. Pectin) 2) Insoluble
Carbohydrates ultimately converted into monosaccharide. The monosaccharides so formed are absorbed
into the intestinal epithelial cells.
o Carbohydrate metabolism begins with digestion in the small intestine where
monosaccharides are absorbed into the blood stream.
o Blood sugar concentrations are controlled by three hormones:
o insulin, glucagon, and epinephrine.
o If the concentration of glucose in the blood is too high, insulin is secreted by the pancreas.
Insulin stimulates the transfer of glucose into the cells. Glucose transporter GLUT.
o *Too much glucose in the cell can cause osmotic imbalance which will disrupt the cell.
o *Glucose 1st transported to the liver (not insulin dependent). Insulin is secreted when glucose
enters into the pancreatic -cell. Glucose as signaling molecule.
In the liver and muscles, most of the glucose is changed into glycogen by the process of glycogenesis
(anabolism). Glycogen is stored in the liver and muscles until needed at some later time when glucose
levels are low. If blood glucose levels are low, then epinephrine and glucogon hormones are secreted to
stimulate the conversion of glycogen to glucose. This process is called glycogenolysis (catabolism).
If glucose is needed immediately upon entering the cells to supply energy, it begins the metabolic process
called glycolysis (catabolism). The end products of glycolysis are pyruvic acid and ATP. Since glycolysis
releases relatively little ATP, further reactions continue to convert pyruvic acid to acetyl CoA and then
citric acid in the citric acid cycle. The majority of the ATP is made from oxidations in the citric acid cycle in
connection with the electron transport chain. During strenuous muscular activity, pyruvic acid is
converted into lactic acid rather than acetyl CoA. During the resting period, the lactic acid is converted
back to pyruvic acid. The pyruvic acid in turn is converted back to glucose by the process called
gluconeogenesis (anabolism). If the glucose is not needed at that moment, it is converted into glycogen
by glycogenesis.
Glucose Transport:
1. You eat causing blood sugar to rise, this is known as the "Fed" state.
2. The liver signals the pancreas to release Insulin, a hormone responsible for utilizing blood glucose.
Insulin-Insensitive: uptake of glucose by the liver, brain and RBC is maximally active in the absence of
insulin, insulin-insensitive. Since glucose reaches the liver before it signals the pancreas to release insulin
it is important that the liver function independently of insulin. -----> prevents hyperglycemia.
Insulin-Sensitive: associated with glucose promoting pancreatic release of insulin.

Pathways of carbohydrate System:

16-02-16 L-12
Overviews of Glucose Metabolism in Selected Tissues:
a= Insulin-Insensitive Transport of Glucose

b= Glycolysis

c= PPP

d= Glycolysis

e= Leaves the Cell as Lactate

a= Insulin-Insensitive Transport of Glucose

b= Glycolysis

c= PPP

d= Glycolysis

f= Metabolism to Acetyl CoA (PDH)

g= Aerobic Energy Metabolism (CO2 End product) - TCA Cycle

a= Insulin-*Sensitized Transport of Glucose b= Glycolysis c= PPP d= Glycolysis

e= Anaerobic Energy Metabolism (Lactate End product) - High Intensity

Exercise

f= Metabolism to Acetyl CoA (PDH)

g= Aerobic Energy Metabolism (CO2 End product) - TCA Cycle

h= Glycogenesis

i= Glycogenolysis
a= Insulin-*Sensitized Transport of Glucose

b= Glycolysis c= PPP

d= Glycolysis f= Metabolism to Acetyl CoA (PDH)

h= Glycogenesis i= Glycogenolysis

j= Lipogenesis

a= Insulin-Insensitive Transport of Glucose b= Glycolysis c= PPP

d= Glycolysis e= Lactate is Transported and Metabolized

f= Metabolism to Acetyl CoA (PDH)

g= Aerobic Energy Metabolism (CO2 End product) - TCA Cycle

h= Glycogenesis i= Glycogenolysis

j= Lipogenesis k= Gluconeogenesis

Hormones in controlling blood glucose level:

Insulin: Secreted from Pancreatic cells (lowers blood glucose level)-
1) Enhances entry of glucose into cells;
2) Enhances storage of glucose as glycogen, or conversion to fatty acids;
3) Enhances synthesis of fatty acids and proteins;
4) Suppresses breakdown of proteins into amino acids, fats of adipose tissue into free fatty acids.
Glucagon secreted from pancreatic -cells raises blood glucose levels.
1) Enhances release of glucose from glycogen;
2) Enhances synthesis of glucose from amino acids
The blood sugar concentration or blood glucose level is the amount of glucose (sugar) present in the blood
of a human or animal. Normally in mammals, the body maintains the blood glucose level at a reference
range between about 3.6 and 5.8 mM (mmol/L) or 64.8 and 104.4 mg/dL.
Three general classes of transport systems:
UNIPORT: systems that transport only one solute.
COTRANSPORT: (obligatory) transport of 2 solutes at the
same time.
Symport: the cotransported solutes go in the same direction
Antiport: the cotransported solutes go in opposite directions
Glucose is polar, therefore is hydrophilic
Facilitated diffusion occurs via 5 glucose transporters.
Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over
a plasma membrane.
Glucose transport in Mammals
GLUTs are integral membrane proteins that contain 12 membrane-spanning helices with both the amino
and carboxyl termini exposed on the cytoplasmic side of the plasma membrane.

22-02-16 L-13
Dietary Carbohydrate Starch, glycogen, Sucrose, Galactose, Lactose
Glycogen More compact (branching after every 8-10 residues and (1-4) linkage.
Starch Less compact (branching after every 20-30 residue and (1-4) linkage.
Breakdown occurs at the same way. Breakdown produces limit dextrin. Digestion stops at branching (1-6
linkage). Debranching enzymes are required for this breakdown. Then the chain is open for amylase to
function. Amylase works on dietary glycogen not on endogenous glycogen.
Dietary glycogenAmylase free glucose
Endogenous Glycogen glycogen phosphorylase glucose-1-P phosphoglucomutase glucose-6-P.
How glucose transporter work?
GLUT proteins transport glucose and related hexoses according to a model of alternate conformation
which predicts that the transporter exposes a single substrate binding site toward either the outside or
the inside of the cell. Binding of glucose to one site provokes a conformational change associated with
transport, and releases glucose to the other side of the membrane. These transport proteins mediate
facilitated transport, that is, they can only transport glucose (or fructose) from areas of high
concentration to areas of lower concentration. The sugar is bound by the protein, a flip-flop mechanism
reverses the membrane direction of the sugar-protein complex, the sugar is released and the protein flips
around once more to initiate a new cycle. Transport activity is dependent upon the sugar concentrations
and the number of transport proteins in the outer cell membrane. In principle the GLUT family can
transport glucose both into and out of cells. In most tissues the internal glucose concentration is quite
low; transport can only proceed from the extracellular area into the cell. In gluconeogenetic tissues
(liver and kidney), intracellular glucose concentration can exceed blood glucose concentration in the
post-absorptive or fasting states. Export of glucose from liver and kidney occurs through GLUT2.
GLUT1 is widely distributed in fetal tissues. In the adult, it is expressed at highest levels in erythrocytes
and also in the endothelial cells of barrier tissues such as the blood brain barrier. However, it is responsible
for the low-level of basal glucose uptake required to sustain respiration in all cells.
GLUT2 is a bidirectional transporter, allowing glucose to flow in 2 directions. Is expressed by renal tubular
cells, small intestinal epithelial cells, liver cells and pancreatic -cells. All three monosaccharides (glucose,
galactose and fructose) are transported from the intestinal mucosal cell into the portal circulation by
GLUT2.
GLUT3 Expressed mostly in neurons (where it is believed to be the main glucose transporter isoform), and
in the placenta.
GLUT4 Found in adipose tissues and striated muscle (skeletal muscle and cardiac muscle).
GLUT5 found in Mucosal surface in small intestine, sperm. Primarily fructose carrier in intestine.
Glucose transport in intestinal epithelial cells
SGLT= Sodium Dependent Glucose Transporter.

Model for glucose transport into erythrocytes by GluT1.

Transporter exists in 2 conformations, T1 with glucose binding site
exposed on outer surface of plasma membrane, and T2, with
binding site exposed on inner surface. D-Glc binding on outside to
stereospecific binding site on T1 conformation triggers
conformational change to T2. Glc is released into cytosol,
triggering conformational change back to T1, ready to pick up
another glucose from the outside. Process is fully reversible, and
as [S]in approaches [S]out, rates of entry and exit become equal.
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Glucose transport by GLUT4 into a myocyte
Diabetes a common metabolic disorder associated with hyperglycemia (increased glucose level, >7mM)
Glucose Homeostasis: maintain normal glucose level (3.5-6.5 mM before meal) by Insulin and glucagon,
1) Glucose uptake & utilization by peripheral tissues (mainly skeletal muscle).
2) Glucose production in the liver.
3) Action of insulin & counter regulatory action of glucagon.
Renal threshold level of blood glucose- 10mM/L kidney can retain glucose will not pass with urine.
Impaired Glucose Tolerance (IGT) Abnormally high level of glucose
First Random Blood Glucose Level is measured. If it is >7.0mmole/L then Fasting Blood Glucose level is
measured (Usually Overnight fasting). If it is >6.0 mmole/L then it is abnormal. So Glucose Tolerance Test
(GTT) is required.
They will take a small sample of blood and a urine sample. After giving a blood sample, you will be asked
to drink an extremely sweet and concentrated solution of glucose within a given amount of time (usually
five minutes). After this, you will be asked to sit in the waiting area until your glucose levels will be tested
again. If you are taking the 50-gram, or one-hour test, they will take your blood sample after one hour. If
you are taking the 75-gram, or two-hour test, they will take a blood sample every hour for two hours. If
you are taking the 100-gram, or three-hour test, they will take a blood sample every hour for three hours.
Diabetics (you will definitely find glucose in urine)
IGT (Glucose in the urine may or may not be found)
Symptoms: PPP=Polyuria (frequent urination), Polydipsia (thirst),
Polyphagia (hunger).
Management: DDD= Diet (take more complex carbohydrates with
the replacement of simple carbs), Discipline, Drug
Type1: No insulin secretion. Type2: Insulin resistance
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Glucose sensitive secretion of insulin from pancreatic beta-cell.

Hypoglycemia, also known as low blood sugar, is when blood sugar decreases to below normal levels. The
glucose level that defines hypoglycemia is variable. Generally, for adults when blood glucose level is
<3.0mM it is considered as severe hypoglycemia but for newborn baby it is <2.0mM. This condition is
more dangerous than hyperglycemia as people often dies of this. Newborn babies are in more prone to it
than adult as they cant share their feeling and it is more frequent in newborn.
Gestational diabetes also known as gestational diabetes mellitus (GDM), is when a woman without
diabetes, develop high blood sugar levels during pregnancy. Babies born to mothers with poorly treated
gestational diabetes are at increased risk of being too large, having low blood sugar or hypoglycemia as
they started to release more insulin in response to high sugar level in mothers blood. A mother's high
blood sugar may increase her risk of early labor and delivering her baby before its due date. Premature
babies, especially those with low birthweights (<2.5kg), who often have limited glycogen stores (sugar
stored in the liver) or an immature liver function.
Symptoms of hypoglycemia in newborn baby:
- Restlessness
- Blue skin color
- Low body temperature
- Sudden stop of breathing
- Poor body ton (less responsive)
Type 1 diabetes or IDDM is the type of diabetes that typically develops in children and young adults. In
type 1 diabetes the body stops making insulin and the blood sugar (glucose) level goes very high.
Treatment to control the blood glucose level is with insulin injections and a healthy diet. Type 1 diabetes
makes up an estimated 510% of all diabetes cases. It occurs at a very early age (12-14).
Diabetes mellitus type 2 or NIDDM is a long term metabolic disorder that is characterized by high blood
sugar, insulin resistance, and relative lack of insulin. Some people are more genetically (insulin receptor
not working properly) at risk than others. Type 2 diabetes makes up about 90% of cases of diabetes. If
blood sugar levels are not adequately lowered, the medication metformin is typically recommended.
Many people may eventually also require insulin injections.