Glycolysis

Presentation

By Charmaine and Joanna

CONTENTS OF THIS PRESENTATION
● Metabolism
● Introduction to Carbohydrate Metabolism
● Glycolysis
● Regulation of glycolysis
● Biomedical Importance of glycolysis+Medical and Pharmaceutical
Implications
● Fate of Pyruvate
What is Glycolysis Aka Embden-Meyerhof-Parnas(EMP) pathway

This is the sequence of reactions that converts glucose into two molecules of
pyruvate in the presence of oxygen(aerobic) and lactate in the absence of
oxygen(anaerobic) with the production of ATP.
MORE INSIGHT…
● Glycolysis is a unique pathway because it can function by utilizing oxygen if available or, it can
function in the total absence of oxygen.
● Glycolysis not only yields pyruvate and ATP(chemical energy) but it also generates intermediates
for other biosynthetic pathways.
● Location—in the cytosol of all cells
● The process of glycolysis consists of 10 enzyme-catalysed reactions. These 10 reactions are
further divided into two phases:
1. Ist phase–Energy requiring/Preparatory phase (Investment)
2. 2nd phase- Energy generating phase (pay off)

● Let's dive into the Investment phase!🙃

 Investment
phase
Aka Energy Requiring phase
Lets see how we prepare for our final product
(pyruvate)!🤑🤗
 1
Reaction 1
Glucose—---------Glucose 6 phosphate
REACTION 1: Phosphorylation of Glucose to Glucose 6 phosphate
● This reaction is an irreversible reaction.
● The 6th carbon of glucose is phosphorylated to yield Glucose 6 Phosphate which is trapped inside the cell due
to its polarity
● The enzymes used are either Hexokinase or Glucokinase (usually used in emergencies,when there is a very
high concentration of glucose(affected by insulin). it has a low affinity for glucose), in the presence of a
cofactor (Magnesium ion)
● This polar phosphate group traps the glucose inside the cell and decreases the concentration of regular
glucose in the cell, which “encourages” more glucose to enter the cell via the GLUT 4 transporters.
● This step costs 1 ATP which provides the necessary phosphate group.
● Hexokinase is also inhibited by its product, glucose 6 phosphate whereas, glucokinase(specific to glucose
only) isn’t,
 2
Reaction 2
Glucose 6 phosphate—------Fructose 6 phosphate
 Reaction 2: Glucose 6 Phosphate is isomerized
● The enzyme phosphoglucose isomerase or phosphohexose isomerase, isomerize the Glucose 6
phosphate to fructose 6 phosphate
● This reaction is reversible.
● Isomerase e
 3
Reaction 3
Fructose 6 phosphate—--------Fructose 1,6 Bisophosphate
 Reaction 3: Phosphorylation of Fructose 6 Phosphate
● This is where the money is!🤑
● This is the rate limiting step of glycolysis and it is catalyzed by PFK-1 (phosphofructokinase-1) which is the
rate limiting enzyme.
● This reaction is irreversible!
● The second and last ATP molecule is invested here, which gives off the phosphate group to Fructose 6
Phosphate on Carbon 1 to yield ADP and Fructose 1,6 Bisphosphate.This commits this molecule to
Glycolysis!
● The enzyme PFK-1 is highly regulated. It is allosterically activated by Fructose 2.6 Bisphosphate ( When the
concentration of fructose-6-phosphate increases in the liver, some of it is converted to F-2,6-BP which is an allosteric
activator of PFK, thus increasing its activity)
● and Increased ADP and AMP molecules and allosterically inactivated by excess ATP(lowers its affinity for
Fructose 6 Phosphate(Inhibition of PFK by ATP results in F6P buildup. F6P exists in equilibrium with G6P so a
high concentration of F6P also leads to a high concentration of G6P. G6P then inhibits hexokinase. So PFK
communicates with hexokinase in this manner to shut down glycolysis) ,) and citrate.
● In cases where a lot of ATP is needed, PFK-2(inhibited by glucagon- a fasting hormone and activated by
insulin)is able to convert Fructose 6 Phosphate to Fructose 2, 6 Bisphosphate.
 4
Reaction 4
Fructose 1,6 bisphosphate—-------Glyceraldehyde 3 phosphate and Dihydroxyacetone
phosphate
 Reaction 4:Cleavage of Fructose 1. 6 Bisphosphate

● A lyase (aldolase) cleaves Fructose 1. 2 Bisphosphate to Glyceraldehyde 3 phosphate and Dihydroxyacetone

Phosphate.
● Basically spitting the 6 Carbon molecule into 2 3 carbon molecules.
● The enzyme aldolase may also be referred to as fructose bisphosphate aldolase.
 5
Reaction 5
Dihydroxyacetone phosphate to Glyceraldehyde 3 phosphate
Reaction 5: Isomerization of Dihydroxyacetone Phosphate

● Initially, Dihydroxyacetone phosphate molecules have a greater percentage compared to Glyceraldehyde 3

Phosphate molecules, but due to continued consumption of Glyceraldehyde 3 Phosphate by the system.
Dihydroxyacetone Phosphate is converted to Glyceraldehyde 3 Phosphate. This occurs due to Le Chatelier's
principle which states that if a stress is applied to a system at equilibrium, the position of equilibrium will shift in
order to reduce the stress.
● In this case, the stress is the high consumption of Glyceraldehyde 3 Phosphate and the change is producing more of
it through isomerization🙃
● This is an irreversible reaction catalyzed by Triose phosphate isomerase.
● This reaction marks the end of the preparatory phase
 Why do some
Mg2+ enzymes work in the
presence of a
Magnesium is an important coherent controller of
glycolysis. Many glycolytic enzymes are sensitive
magnesium ion?
to Mg2+, particularly those whose principle
function is to facilitate the transfer of high energy
phosphate (kinases). Its functions include: it being
a cofactor(a non-protein that binds to either the
enzymes or the substrate and helps its activity) for
these enzyme controlled reactions and it critically
stabilizing enzymes including those involved in
many ATP generating reactions.
 Biomedical importance
03
Haemolytic anaemia Type 1 diabetes
01 (insulin dependent) here there is nsulin deficiency (a decrease in the circulating
Defects in glycolysis (e.g concentration of insulin), Without insulin, cells are unable to use glucose as fuel and they
will start malfunctioning. In diabetes type 1, the pancreas does not make insulin, because
pyruvate kinase the body's immune system attacks the islet cells in the pancreas that make insulin. So if
deficiency)lead to low oxygen there is no insulin, the Glut transporters are not "activated" to go to the cell membrane and
create channels for glucose to enter the cell and start glycolysis to give energy. It is the
production in RBC which beta cells of the pancreas that make insulin.
results in their destruction
ANALYSIS 04
02 Type 2 diabetes -remember the mnemonic for glut transporters, so the reason why the liver,brain and red blood cells are
not stimulated by insulin in because of the GLUT transporters in these areas, they are not sensitive to
insulin hence there organs are also insensitive to insulin. How does insulin stimulate these muscles and
adipose tissue?, well it causes the glut transporters to move to the cell membrane and then form
In diabetes type 2, the pancreas makes less insulin than used to, channels that allow glucose to enter.In both cases, the glut-4 transporters are not adequately transported
from the cytoplasm to the cell membrane to allow glucose entry into the muscle cell.These
and your body becomes resistant to insulin.a decrease in the abnormalities lead to alterations in the metabolism of carbohydrates, lipids, ketones, and amino acids;
the central feature of the syndrome is hyperglycemia.
response of peripheral tissues to insulin. So even if insulin is
present the GLUT transporters are not stimulated to allow Insulin can be administered to individuals with Type 1 Diabetes to allow glucose enter the cell and
glucose to enter.This leads to high levels of blood glucose. undergo metabolic processes. Insulin can also be given in some cases of Type 2 diabetes
 Pay-off
phase!
Aka Energy giving phase
Lets see how we get value for our
investment!🤑🤗
NB-The action of aldolase (it is a lyase, thus it catalyzes the cleavage of a carbon-carbon bond) splits the 6 carbon
Fructose 1,6 bisphosphate into two 3 Carbon molecules. Hence from here on we are dealing with propanes🙃
After the 5th reaction, we are now working in 2’s because we have now produced 2 Glyceraldehyde -3 Phosphate
molecules.
 6
Reaction 6
Glyceraldehyde 3 phosphate—---- 1, 3 bisphosphoglycerate
REACTION 6 : Oxidation of Glyceraldehyde 3 Phosphate to 1, 3 Bisphosphoglycerate
● This is a reversible reaction.
● This is the first step of us stealing electrons from “glucose”!
● This reaction is catalysed by the enzyme Glyceraldehyde 3 phosphate dehydrogenase
*NB:Dehydrogenases are oxidoreductase enzymes. They catalyze the oxidation of a substrate through the
transfer of hydrogen (hydride) to an acceptor..
● With that being said, Glyceraldehyde 3 phosphate dehydrogenase works by making NAD+(electron carrier)
react with Glyceraldehyde 3 phosphate by letting it rip off a hydride (oxidizes) from Glyceraldehyde 3-
phosphate , leading to the formation of NADH (NAD+ is reduced)
Hydride= a proton + two electrons
● Initially, the first phosphate is on the 3rd carbon of Glyceraldehyde 3 phosphate, so when we add another
inorganic phosphate phosphate to the 1st carbon it is now called 1,3 Bisphosphate.
*But why “bis-” and not “bi-”🧐, because the attachments are not close to each other!
● Coenzyme (non-protein organic molecule that helps enzyme activity) NAD+ is a biological oxidizing agent
that converts C-H bonds to C-O bonds. In the process, NAD+ is reduced to NADH + H+.
 7
Reaction 7
1,3 bisphosphoglycerate—----3 phosphoglycerate
REACTION 7: Dephosphorylation from 1,3 bisphosphoglycerate to ADP
● This is also a reversible reaction.
● Here, we are getting rid of the phosphate we just added😂
● It is catalysed by the enzyme phosphoglycerate kinase
What does a kinase do🤔?- It phosphorylates substances, in this case ADP.
● Essentially this reaction is about taking a phosphate from carbon 1 of 1,3
Bisphosphoglycerate and giving it to ADP to form ATP.
● Remember we have two 1,3 bisphosphoglycerate molecules, so we take two ADP’s and
make 2 ATP’s
● The molecule remaining is now 3 Phosphoglycerate!
● ATP formation by direct phosphate group transfer from a substrate such as 1,3 BPG is
referred to as substrate level phosphorylation.
●
NB: Asernate
Asernate (asernic acid)is structurally similar to phosphate and thus is can also
participate in Step 6 in place of the inorganic phosphate. When it does, it reacts
with Glyceraldehyde 3 Phosphate and leads to the formation of an unstable
intermediate that is rapidly hydrolyzed to 3 Phosphoglycerate a product of Step
7, this means that we jump the step of ATP production and now only 2
molecules will be produced at the end of the reaction. With consideration of
our capital this will lead to a loss in our glycolysis business😭
 8
Reaction
3 phosphoglycerate—----- 2 phosphoglycerate
REACTION 8: CONVERSION OF 3-PHOSPHOGLYCERATE TO 2-
PHOSPHOGLYCERATE
● This is a reversible reaction.
● The phosphate group on carbon 3 of 3-phosphoglycerate is transported to the second carbon to
give 2-phosphoglycerate
● This reaction is catalysed by the enzyme phosphoglycerate mutase.
What does a mutase do🤔?- A mutase falls under the class of isomerase enzymes, it catalyzes the
movement of a functional group from one position to another within the same molecule.
 9
Reaction 9
2- phosphoglycerate—----Phosphoenolpyruvate
REACTION 9: Dehydration of 2 Phosphoglycerate to Phosphoenolpyruvate
● This reaction is reversible
● In the reaction, the enzyme enolase adds an enol to 2-Phosphoglycerate to form Phosphoenolpyruvate.
● Water is given out as a a result of this reaction (Dehydration reaction)
● Phosphoenolpyruvate is the first form of pyruvate obtained (an enol), it is later converted to the ketone form
(pyruvate) which is more stable
● NB- the enzyme enolase catalyzes the same reaction in gluconeogenesis
 Pharmaceutical
implication
The enzyme enolase is inhibited by
fluoride. To prevent ongoing glycolysis in
a patient’s blood samples collected for
sensitive glucose tolerance tests, blood
is collected in tubes containing fluoride
 10
Reaction 10
Phosphoenolpyruvate—-------Pyruvate
REACTION 10: Transfer of the phosphoryl group from Phosphoenolpyruvate to ADP
● This is the 3rd irreversible reaction in glycolysis
● In this reaction, phosphates are taken again, thus the enzyme has to be a kinase!
● Pyruvate kinase(highly regulated) catalyzes the transfer of phosphate from phosphoenolpyruvate to ADP,
forming ATP, thus leaving pyruvate (a 3 carbon molecule).
● Here 2 ADP molecules are phosphorylated thus 2 ATP molecules are made.This is the second substrate level
phosphorylation stage of glycolysis.
● Phosphoenolpyruvate requires 2 cofactors whicha re magnesium ion and potassium ion.
Regulation of Pyruvate Kinase
● NB: all the 3 irrevesible reactions are exergonic, The enzymes involved function
as “valves”, regulating the flow of carbon through glycolysis.The rates of these
steps are limited not by the substrate but by the activity of the enzymes
● Pyruvate kinase is allosterically inactivated by excess ATP
● This enzyme is also non competitively inhibited by alanine (it can bind to the free
enzyme or the ES complex)
 PRODUCTS OF GLYCOLYSIS
NB: in total, 8 molecules of ATP are produced from a single
molecule of glucose

4 ATP’s 2 Pyruvate 2 NADH+H+

Due to the action of kinases in steps 7 and
10. ADP was phosphorylated into ATP. To
calculate our “profit” we subtract what we
invested from what we got. We invested 2
ATP’s in steps 1 and 3, thus our “profit” is 2
ATP molecules
Produced in the final
step.It’s fate will be
discussed shortly!🙃
This will be involved in creating
lactate. Produced in Step 6.It
creates an acidic environment. It
will also be used in the later stages
of cellular respiration.
FATE OF
PYRUVATE
WHAT HAPPENS TO THIS
PRODUCT🧐?
Let's take a look…
Under aerobic Pyruvate is oxidised to acetate which enters
to citric acid cycle where it is then is then
conditions oxidised to Carbon dioxide and Water.

NADH formed is reoxidised by passage of

oxygen in mitochondrial respiration.
 Reduction of Pyruvate to give NAD+

Here, NADH generated by glycolysis cannot be oxidised by oxygen.

This failure to generate NAD+ leaves the cell with no electron acceptor for the
oxidation of glyceraldehyde 3 phosphate (step 6).

Under NAD+ is generated by the reduction of pyruvate to lactate, catalyzed by the

enzyme lactate dehydrogenase. It is this reaction that allows glycolysis to occur in
anaerobic the absence of oxygen.

In red blood cells, under aerobic conditions, glycolysis always ends in lactic acid
conditions production.

E.g in strenuously Other tissues that normally derive their energy from glycolysis and produce lactate
include brain, renal medulla, testes, GIT and retina of eyes.
active muscles
Lactate formed by active muscles is carried in the blood to the liver where its
converted to glucose during the recovery phase

Acidification that results from ionization of lactic acid in muscle and blood causes
pain and limits exercise
 Biomedical importance cont..

Lactic Acidosis
Lactic Acidosis- can be due to several causes of improper
utilization of lactate.Lactic acidosis is a type of metabolic
acidosis that occurs when lactic acids build up in your blood.
Your body produces more lactate when your tissues are deprived
of oxygen. Lactate can also build up if your livers and kidneys
aren't able to metabolize it efficiently
 Importance of Phosphorylated
intermediates
1. 2. 3.
Because the P group is
ionized at pH 7 and has Binding of P groups to the
charge and because plasma P groups are
active sites of enzymes
membranes are not necessary for chemical
permeable to the charged provides binding energy
energy. Energy is
molecules, the phosphorylated which decreases the
molecules can not pass the released after the
activation energy and
membranes. They are trapped hydrolysis of ATP.
in the cell. increase the specificity of
enzyme-catalyzed reactions.
 TABLE OF CONTENTS

1 2 3
ABOUT COLORS PINK CONTENT OVERVIEW
}A variety of D-hexoses, including fructose,
You can describe the You can describe the You can describe the
topic of the section here mannose, and
topic of the section here galactose, can
topic of the be here
section funneled in
glycolysis.
}
THANK YOU!