Metabolism of Glucose

Awotunde O.S (PhD)

Senior Lecturer
Department of Biochemistry HMS, IUIU.
Objectives
• Fate of pyruvate.
• Energy yield of glucose metabolism.
• Feeder Pathway.
 Review on glycolysis
• Glycolysis = ( Greek word- sugars and dissolution)
breakdown of sugars; glucose, fructose, galactose,
mannose etc.
• It occurs not only in animals and plants but also in a
great range of many microorganisms.
 Fate of the intermediates in glycolysis go?

 G-6-P goes off to make the ribose for nucleotides

 F-6-P -amino sugars-glycolipids and glycoproteins
 G-3-P/DHAP-lipids
 3PG-serine
 PEP-aromatic amino acids, pyrimidines, asp and asn
 Pyruvate-alanine

This pathway not only important in glucose metabolism--generates

intermediates for other important building blocks.
G-6-P = glucose 6 phosphate, F-6-P = fructose 6 phosphate, G-3-P = glyceraldehyde 3 phosphate, DHAP =
dihydryoxacetonephosphate, 3PG = phosphoglyceraldehyde, Pyr = pyruvate, PEP- phosphoenol pyruvate,
Asn = Asparagine, ASP Aspartic acid
 Fates of pyruvate (end product of glycolysis)

1. Alcohol fermentation (to form Ethanol and carbon dioxide)

- anaerobic conditions.
2. Oxidation of Pyruvate to Acetyl CoA ( acetyl-coenzyme A)
– occurs in animals, plants and others)- aerobic conditions .
Then to citric acid cycle where Acetyl CoA is oxidized to
carbon dioxide and water ( In animals and plants).
3. Reduction of Pyruvate to Lactate (mammals and others). Its
called anaerobic glycolysis.
Its is the important source of energy in very intense physical
activity.
The metabolic fate of pyruvate
Fate on pyruvate
 Cori cycle

oxaloacetate
 (a ) Reduction of pyruvate to Ethanol
( microorganisms)

 A two step process:

 1) Pyruvate decarboxylase requires thiamine pyrophosphate
TPP as a cofactor.
 2) Alcohol dehydrogenase requires Zn+2 as a cofactor.
 The decarboxylation of pyruvate by pyruvate dehydrogenase in
yeast and other microorganisms but not in humans.
 The enzyme requires thiamine pyro-phosphate as coenzyme
and catalyzes the reaction as pyruvate dehydrogenase.
 The product is acetaldehyde.
 The acetaldehyde is reduced by NADH in presence of Alcohol
dehydrogenase into Ethanol.
Alcoholic ( Ethanol) fermentation
 (b) Homolactic fermentation: conversion of pyruvate
to lactate

O LDH lactate H O
O O -
HR HS O O-
dehydrogenase
NH2 NH2
C O + HO C H +
+ H+
N CH3 N
CH3
R R
Pyruvate NADH L-Lactate NAD+

Mammals have two different types of enzymes: Isozymes

M type for muscle
H type for heart
 Lactate dehydrogenase is a tetramer H4 has a low
Km for pyruvate and is allosterically inhibited by
high concentrations of pyruvate.
M4 has a higher Km for pyruvate and is not
allosterically regulated
Although all five types can exist, H4, H3M, H2M2
HM3, M4.
The M predominates in anaerobic muscle tissues
which favor the formation of lactate while the H4
form predominates in aerobic tissues like heart where
the formation of pyruvate from lactate is preferred
 Pro-R hydride is
transferred from C4 of
NADH to C2 of pyruvate
with the concomitant
transfer of a proton from
His 195
All muscle lactate is transferred to
the liver where it is turned back to
glucose- gluconeogenesis
Cori Cycle
Lactate formation cont’d
 Lactate is the End product of glycolysis in anaerobic
microorganisms that carry out lactate fermentation.

 Lactic acid formed from sugars by lactic acid bacteria

causes the souring of milk.
 Also in eukaryotes, lactate is the end product for anaerobic
glycolysis.

 Lactate is the major fate for pyruvate in lens and cornea of

the eye, testes, leukocytes, kidney medulla and RBCs
( poorly vascularized and or lack mitochondria).
 Energetics of Fermentation

DG'
Glucose 2lactate + 2H+ -196 kJ• mol-1 of glucose
Glucose 2CO2 + 2ethanol -235 kJ• mol-1 of glucose

Formation of 2ATP +61 kJ• mol-1 of glucose

equals 31% and 26% efficient for energy conservation
Under physiological conditions this efficiency
approaches 50%
 ( c ) Oxidative decarboxylation of
pyruvate
It is an important pathway in tissues with high oxidative
capacity e.g. cardiac muscle.

The enzyme required is pyruvate dehydrogenase which

is a complex enzyme.
Pyruvate dehydrogenase irreversibly converts pyruvate
into acetyl CoA.
Importance of acetyl CoA.
1. Major fuel for TCA cycle
2. The building block for fatty acid synthesis.
Energy Balance Sheet for the Oxidation of Glucose via Glycolysis

Gains: Net Gain: Losses:

4 ATP 2ATP
+ 2 ATP Glucose
2 pyruvate
Phosphate
2 NADH + H+
NAD+ (recycled)

Mitochondria for
further oxidation via
the TCA/Krebs cycle
 (d) Carboxylation of pyruvate to oxaloacetate

• The enzyme is pyruvate carboxylase

• This is a biotin- dependent reaction
• Importance
• 1. Replenishes the TCA cycle
• 2. Provides substrate for gluconeogenesis
Summary of TCA cycle
Oxidation of pyruvate via the TCA/Krebs/Citric Acid Cycle
 Pyruvate
CO2 NAD+
NADH
Acetyl CoA

•All compounds are

tricarboxylic acids
•Carbons from glucose
are shown in red
•Carbons from glucose
are lost as CO2
(decarboxylation)
•Several NADH + H+
are generated via
oxidation of
intermediates
•One high energy
phosphate compound
(GTP)is produced
Net Energy Yield from the Oxidation of Pyruvate via the TCA cycle

From Glycolysis:
+2NADH +2ATP

From TCA: + +

+2FADH +8NADH +2GTP

ETC:
3ATP/NADH
2ATP/FADH Do you know why?

+4ATP +30ATP
+38ATP TOTAL
Complex IV
• reduction of oxygen
• cytochrome oxidase
• cyt a+a3 red ---> oxidized state
• oxygen ---> water
• 2 H+ + 2 e- + ½ O2 -- 2 H2O
• transfers e- one at a time to oxygen
• Pumps 2H+ out
• Total of 10 H+ / NADH
• Total of 6 H+ / FADH2
Totals
• Proton gradient created as electrons transferred to oxygen forming
water
10 H+ / NADH
6 H+ / FADH2
Electron Transport Chain
Generation of ATP
• Proton dependant ATP synthetase
• Uses proton gradient to make ATP
• Protons pumped through channel on enzyme
• From intermembrane space into matrix
• ~4 H+ / ATP
• Called chemiosmotic theory
Generation of ATP
• Proton dependant ATP synthetase
• Uses proton gradient to make ATP
• Protons pumped through channel on enzyme
• From intermembrane space into matrix
• ~4 H+ / ATP
• Called chemiosmotic theory
Question
How many ATP is generated from the whole process from substrate
level oxidative phosphrylation?
Totals
NADH
10 H+ X 1 ATP = 2.5 ATP
4 H+

FADH2
6 H+ X 1 ATP = 1.5 ATP
4 H+
Total ATP from mitochondrial matrix
• Pyruvate dehydrogenase
• NADH ……………………………….2.5 ATP
• Krebs
3 NADH X 2.5 ATP/NADH ……….7.5 ATP
FADH2 X 1.5 ATP / FADH2……….1.5 ATP
GTP X 1 ATP / GTP ……………..1.0 ATP
(from a separate reaction)

Total …………….12.5 ATP

(Per glucose = X 2 = 25 ATP)
What about NADH from glycolysis?
• NADH made in cytosol
• Can’t get into matrix of mitochondrion
• 2 mechanisms
• In muscle and brain
• Glycerol phosphate shuttle
• In liver and heart
• Malate / aspartate shuttle
Glycerol Phosphate shuttle

• http://courses.cm.utexas.edu/jrobertus/ch339k/overheads-3/ch19_glycerol-shuttle.jpg
Glycerol phosphate shuttle
• In muscle and brain
• Each NADH converted to FADH2 inside mitochondrion
• FADH2 enters later in the electron transport chain
• Produces 1.5 ATP
Total ATP per glucose in muscle and brain
• Gycerol phosphate shuttle
• 2 NADH per glucose - 2 FADH2
• 2 FADH2 X 1.5 ATP / FADH2……….3.0 ATP
• 2 ATP in glycoysis ……………………2.0 ATP
• From pyruvate and Krebs
• 12.5 ATP X 2 per glucose ……………..25.0 ATP

Total = 30.0 ATP/ glucose

Malate – Aspartate Shuttle

• http://courses.cm.utexas.edu/emarcotte/ch339k/fall2005/Lecture-Ch19-2/Slide14.JPG
Malate – Aspartate Shuttle
in cytosol
• In liver and heart
• NADH oxidized while reducing oxaloacetate to malate
• Malate dehydrogenase
• Malate crosses membrane
Malate – Aspartate Shuttle
in matrix
• Malate reoxidized to oxaloacetate
• Malate dehydrogenase
• NAD+ reduced to NADH
• NADH via electron transport yields 2.5 ATP
Total ATP per glucose in liver and heart
• Malate – Aspartate Shuttle
• 2 NADH per glucose - 2 NADH
• 2 NADH X 2.5 ATP / NADH…………5.0 ATP
• 2 ATP from glycolysis………………..2.0 ATP
• From pyruvate and Krebs
• 12.5 ATP X 2 per glucose ……………..25.0 ATP

Total = 32.0 ATP/ glucose

Summary
• Total ATP / glucose
• Muscle and brain 30.0 ATP
• Uses glycerol phosphate shuttle

• Heart and liver 32.0 ATP

• Uses malate aspartate shuttle
Feeder Pathway of glycolysis
• A feeder pathway for glycolysis is pathway that basically feeds glucose
or another starting material into the glycolysis pathway
Starting materials
In glycolysis most carbohydrates except glucose are transformed into
one of the glycolytic intermediates. Listed below are the most
significant:
• 1.      The storage polysaccharides (glycogen and starch)
• 2.      The disaccharides (maltose, lactose, trehalose and sucrose)
• 3.      The monosaccharides (fructose, mannose and galactose)

• Note: the "starting material" may or may not enter the pathway at the
beginning.
(Metabolism of Polysaccharide)
Glycogen Phosphorylase
• Glycogen Phosphorylase removes the terminal glucose units of
glycogen (a similar enzyme works on starch). The enzyme uses
pyridoxal phosphate as a cofactor and relies on an attack by an
inorganic phosphate. Once this phosphate attacks the bond between
successive glucose residues, the terminal one is removed with a
phosphate ester. This phosphate ester preserves some of the energy
initially stored in the chain. The enzyme will continue in this manner,
taking successive glucose molecules off the end of the chain, until it
reaches four residues from a branch point.
Glucotransferase
• Glucotransferase works at the branch point in coordination with
glycogen phosphorylase to create glucose-1-P.
• But the aim is Glu-6-P.
Phosphoglucomutase
• Phosphoglucomutase converts 1-P to 6-P
• This allows the newly-freed glucose molecules to go through
glycolysis.
• Phosphoglucomutase requires glucose-1,6-bP as a cofactor.
Metabolism of fructose Pathway
• There are two routes in which fructose can be metabolized:
• 1.      In adipose tissue
• 2.      In the liver
• Let’s see how in each case.
• In adipose tissue: Fructose is phosphorylated to Fructose-6-phosphate
that enters into glycolysis by muscle and kidney enzyme Hexokinase
• In the liver: Fructose is metabolized by the Fructose-1-phosphate
pathway by the enzyme Glucokinase.
Monosaccharides
• Monosaccharides are not simply being broken down and fed into the
beginning of the cycle but are converted and then plugged in
Hexokinase
• It carries out the phosphorylation of glucose that enters the cell.
• Using Mg2+ as a cofactor and ATP as a phosphate source, it can
phosphorylate fructose (another hexose) to produce Fru-1-P.
• Note: Fru-1-P is a molecule which is an intermediate in glycolysis, and
the cycle can simply go from there.
Fructokinase, Fru-1-Phosphate aldolase, and
triose kinase
• Hexokinase is not the only enzyme that can act on fructose with ATP,
• In the liver, Fructose and ATP are recombined to again form Fru-1-P
and ADP.
• Then Fru-1-P aldolase forms glyceraldehyde and dihydroxyacetone
and triose kinase acts and the cycle proceeds.
Metabolism of Galactose Pathway

• Galactose enters glycolysis via the galactose-glucose interconversion

pathway which is a 4 step reaction sequence. See image below of this
pathway.
• Begins with galactose and ends with Glu-1-P.
• It uses UDP (Uridine diphosphate) as a cofactor,
Phosphomannose isomerase
• Begings with Man-6-P,
• Phosphomannose isomerase can create Fru-6-P in a single bound.
Disaccharides
• They can also be fed into the pathway.
• The dissacharide breakdown
• Lactase- breaks down lactose,
• Sucrase-breaks down sucrose, etc
• The resulting monomers are acted upon by enzymes as discussed
above.
 Sample questions

(A)1. Name three important routes taken by pyruvate after glycolysis.

State the conditions, enzymes involved and the product formed for each
route taken.

2. Hexoses other than glucose can be -------------------- and

-------------------into intermediates that enter glycolytic pathways. Fill in the
spaces. Also name these hexoses and the intermediate compounds.

3.--------------------is the major fate for pyruvate in lens and cornea of the
eye, testes, leukocytes, kidney medulla and RBCs because ------------------
Sample questions cont’d
4. Name the sugars that can be phosphorylated and
converted into intermediates of the glycolytic pathway .
 Also name the intermediates that feed the glycolytic
pathway in the table below.
Sugar Intermediate that enters the
glycolytic pathway

6. For each of the hexoses given, name the enzymes

involved in their metabolism and the final products formed.
b. Identify and describe the different feeder metabolic pathways seen in the
diagram