Digestion glycolysis

Khalid Hussain
Khalid HussainProfessor um University of the Punjab
Digestion and Absorption of
carbohydrates
 CHO taken in diet are:
 polysaccharides,
 disaccharides
 monosaccharides
 These are supplied from external sources,
hence called exogenous CHO
 These may be
 digestible or
 indigestible
Absorption and digestion of carbohydrates
Digestion of CHO takes place in;
 mouth
stomach
Intestine
 Absorption takes place form small the
intestine
Digestion: Mouth
At slightly acidic pH, salivary amyalse (ptyalin) acts
on starch, which is converted into maltose and
isomaltose
The enzyme get inactivated in stomach
2(C6H10O5)n + nH2O nC12H22O11
starch maltose and isomaltose
Stomach
• HCl can cause hydrolysis of starch into maltose and
isomaltose and that of maltose to glucose but the
reaction is of little significance inside stomach
Small intestine
In the small intestine pancreatic amylase converts 87%
starch to maltose and isomaltose and 13 % glucose
(C6H10O5)n + nH2O nC6H12O6
Disaccharides present in brush border of
epithelial cells are lactose, sucrose, maltose
and isomaltose
Lactase, sucrase, maltase and isomaltase
hydrolyze disaccharides into their
components as:
Sucrose into glucose and fructose
Maltose into glucose
Lactose into glucose and galactose
Absorption
Glucose (80-85%) and few disaccharides
are absorbed from small intestine through
capillaries of intestinal mucosa
The order of ease of absorption of
monosaccharides is galactose > glucose >
fructose > mannose > pentose
Mechanism of absorption
Simple diffusion
Active transport
Factors upon which absorption of CHO depends
are given as:
Physical factors (how long food stays in intestine)
Hormone
thyroid hormones increase the rate of
absorption
hormones of adrenal cortex facilitate
absorption
Facts about absorption of
monosaccharides
Chemical nature of the most actively
transported monosaccharides has following
features in the choice of the carrier:
Presence of 6 and more carbon atoms
D-pyarnose structure
Intact OH at carbon 2
Glycolysis
Glucose and glycogen are broken down in the body
by a complex chain of reactions catalyzed by many
enzymes
There are many metabolic pathways by which
glucose can be utilized in the body; the most
important one is the Embden-Meyerhof pathway
followed by citric acid cycle
Glycolysis is of two types:
anaerobic and
aerobic
Anaerobic glycolysis
It takes place in cytosol (extra-mitochondrial)
Glucose is broken into two molecules of pyruvic
acid, which is then converted into lactic acid by
utilizing NADH/H+
It can not continue indefinitely;
 lactic acid lowers the pH to a level that is not suitable
for cellular function
On the other hand NADH/H+
becomes unavailable, if
aerobic metabolism remains suspended for a long time
Aerobic glycolysis
In the presence of oxygen the pyruvic acid is
formed in the same way as anaerobic
glycolysis but it does not give rise to lactic
acid
Pyruvic acid is converted into acetyle-CoA
which then enters in citric acid cycle
Reactions of citric acid cycle occur in
mitochondria
Main Features
The oldest of the Pathways
Occurs in Soluble Phase of Cytoplasm
(Cytosol)
Anaerobic Phase Energy
Generates ATP
Produces Pyruvate/Lactate
Produces Many Important Intermediates of
other Pathways
Relationship to Other Pathways
TCA Cycle
Gluconeogenesis (in Liver and Kidney)
Hexose Monophosphate Shunt (HMP)
Metabolism of other Sugars, e.g., Fructose and
Galactose
Metabolism of certain amino acids
Lipid metabolism
Glycoprotein Synthesis
Transport of glucose into the cell
Transportation of glucose is mediated by two types of
systems that are given as follows:
1- Insulin-independent transport system
Hepatocytes, erythrocytes and brain cells do not need
insulin for the entry of glucose. Transport occurs by a
protein, which is an oligomer (MW 200,000)
containing 4 sub-units of equal size
Transport of glucose into the cell
2- Insulin-dependent transport system
It occurs in muscles and adipose tissue cells. The
binding of insulin to the receptors enhances the
transport of glucose into cell by causing
o migration of glucose transport protein from
microsomes to plasma membrane
o and by increasing transport capacity of the transport
proteins
Glycolysis (Embden-Meyerhof pathway)
Glycolysis takes place in the cytosol of the cells
Glucose enters the glycolysis pathway by
conversion to glucose-6-phosphate, which is
initially an energy consuming step; energy input
corresponding to one ATP
H O
OH
H
OHH
OH
CH2OPO3
2−
H
OH
H
1
6
5
4
3 2
glucose-6-phosphate
H O
OH
H
OHH
OH
CH2OH
H
OH
H H O
OH
H
OHH
OH
CH2OPO3
2−
H
OH
H
23
4
5
6
1 1
6
5
4
3 2
ATP ADP
Mg2+
glucose glucose-6-phosphate
Hexokinase
1. Hexokinase catalyzes:
Glucose + ATP  glucose-6-P + ADP
The reaction involves nucleophilic attack of C-6
hydroxyl of glucose by P of the terminal phosphate
of ATP. ATP binds to the enzyme as a complex
with Mg++
Mg++
interacts with negatively charged phosphate
oxygen atoms, providing charge compensation &
promoting a favorable conformation of ATP at the
active site of the Hexokinase enzyme
N
N
N
N
NH2
O
OHOH
HH
H
CH2
H
OPOPOP
−
O
O
O
−
O
−
O O
O
−
adenine
ribose
ATP
adenosine triphosphate
The reaction catalyzed by Hexokinase is highly
spontaneous
A phosphoanhydride bond of ATP (~P) is cleaved
The phosphate ester formed - glucose-6-phosphate
- has a lower ∆G of hydrolysis
H O
OH
H
OHH
OH
CH2OH
H
OH
H H O
OH
H
OHH
OH
CH2OPO3
2−
H
OH
H
23
4
5
6
1 1
6
5
4
3 2
ATP ADP
Mg2+
glucose glucose-6-phosphate
Hexokinase
2. Phosphoglucose Isomerase catalyzes:
glucose-6-P (aldose)  fructose-6-P
(ketose)
H O
OH
H
OHH
OH
CH2OPO3
2−
H
OH
H
1
6
5
4
3 2
CH2OPO3
2−
OH
CH2OH
H
OH H
H HO
O
6
5
4 3
2
1
glucose-6-phosphate fructose-6-phosphate
Phosphoglucose Isomerase
3. Phosphofructokinase catalyzes:
fructose-6-P + ATP  fructose-1,6-bisP + ADP
The Phosphofructokinase reaction is the rate-limiting
step of Glycolysis.
CH2OPO3
2−
OH
CH2OH
H
OH H
H HO
O
6
5
4 3
2
1 CH2OPO3
2−
OH
CH2OPO3
2−
H
OH H
H HO
O
6
5
4 3
2
1
ATP ADP
Mg2+
fructose-6-phosphate fructose-1,6-bisphosphate
Phosphofructokinase
4. Aldolase catalyzes:
fructose-1,6-bisphosphate 
dihydroxyacetone-P + glyceraldehyde-3-P
The reaction is an aldol cleavage, the reverse of an aldol
condensation.
6
5
4
3
2
1CH2OPO3
2−
C
C
C
C
CH2OPO3
2−
O
HO H
H OH
H OH
3
2
1
CH2OPO3
2−
C
CH2OH
O
C
C
CH2OPO3
2−
H O
H OH+
1
2
3
fructose-1,6-
bisphosphate
Aldolase
dihydroxyacetone glyceraldehyde-3-
phosphate phosphate
Triosephosphate Isomerase
5. Triose Phosphate Isomerase catalyzes:
dihydroxyacetone-P  glyceraldehyde-3-P
6
5
4
3
2
1CH2OPO3
2−
C
C
C
C
CH2OPO3
2−
O
HO H
H OH
H OH
3
2
1
CH2OPO3
2−
C
CH2OH
O
C
C
CH2OPO3
2−
H O
H OH+
1
2
3
fructose-1,6-
bisphosphate
Aldolase
dihydroxyacetone glyceraldehyde-3-
phosphate phosphate
Triosephosphate Isomerase
The ketose/aldose conversion involves acid/base
catalysis, and is thought to proceed via an enediol
intermediate, as with Phosphoglucose Isomerase.
C
C
CH2OPO3
2−
O
C
C
CH2OPO3
2−
H O
H OH
C
C
CH2OPO3
2−
H OH
OH
H
H OH H+
H+
H+
H+
dihydroxyacetone enediol glyceraldehyde-
phosphate intermediate 3-phosphate
Triosephosphate Isomerase
C
C
CH2OPO3
2−
H O
H OH
C
C
CH2OPO3
2−
O OPO3
2−
H OH
+ Pi
+ H+
NAD+
NADH 1
2
3
2
3
1
glyceraldehyde- 1,3-bisphospho-
3-phosphate glycerate
Glyceraldehyde-3-phosphate
Dehydrogenase
6. Glyceraldehyde-3-phosphate Dehydrogenase
catalyzes:
glyceraldehyde-3-P + NAD+
+ Pi

1,3-bisphosphoglycerate + NADH + H+
C
C
CH2OPO3
2−
O OPO3
2−
H OH
C
C
CH2OPO3
2−
O O−
H OH
ADP ATP
1
22
3 3
1
Mg2+
1,3-bisphospho- 3-phosphoglycerate
glycerate
Phosphoglycerate Kinase
7. Phosphoglycerate Kinase catalyzes:
1,3-bisphosphoglycerate + ADP 
3-phosphoglycerate + ATP
C
C
CH2OH
O O−
H OPO3
2−
2
3
1
C
C
CH2OPO3
2−
O O−
H OH2
3
1
3-phosphoglycerate 2-phosphoglycerate
Phosphoglycerate Mutase
8. Phosphoglycerate Mutase catalyzes:
3-phosphoglycerate  2-phosphoglycerate
Phosphate is shifted from the OH on C3
to the OH on C2.
9. Enolase catalyzes:
2-phosphoglycerate  phosphoenolpyruvate +
H2
O
This dehydration reaction is Mg++
- dependent.
C
C
CH2OH
O O−
H OPO3
2−
C
C
CH2OH
−
O O−
OPO3
2−
C
C
CH2
O O−
OPO3
2−
OH−
2
3
1
2
3
1
H+
2-phosphoglycerate enolate intermediate phosphoenolpyruvate
Enolase
10. Pyruvate Kinase catalyzes:
phosphoenolpyruvate + ADP  pyruvate + ATP
C
C
CH3
O O−
O2
3
1
ADP ATP
C
C
CH2
O O−
OPO3
2−
2
3
1
phosphoenolpyruvate pyruvate
Pyruvate Kinase
This phosphate transfer from PEP to ADP is spontaneous
 PEP has a larger ∆G of phosphate hydrolysis than ATP
 Removal of Pi
from PEP yields an unstable enol, which
spontaneously converts to the keto form of pyruvate
C
C
CH3
O O−
O2
3
1
ADP ATPC
C
CH2
O O−
OPO3
2−
2
3
1
C
C
CH2
O O−
OH2
3
1
phosphoenolpyruvate enolpyruvate pyruvate
Pyruvate Kinase
Hexokinase
Phosphofructokinase
glucose Glycolysis
ATP
ADP
glucose-6-phosphate
Phosphoglucose Isomerase
fructose-6-phosphate
ATP
ADP
fructose-1,6-bisphosphate
Aldolase
glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
Glyceraldehyde-3-phosphate
Dehydrogenase
Phosphoglycerate Kinase
Enolase
Pyruvate Kinase
glyceraldehyde-3-phosphate
NAD+
+ Pi
NADH + H+
1,3-bisphosphoglycerate
ADP
ATP
3-phosphoglycerate
Phosphoglycerate Mutase
2-phosphoglycerate
H2O
phosphoenolpyruvate
ADP
ATP
pyruvate
C
C
CH3
O−
O
O
C
HC
CH3
O−
OH
O
NADH + H+
NAD+
Lactate Dehydrogenase
pyruvate lactate
E.g., Lactate Dehydrogenase catalyzes reduction of the keto in
pyruvate to a hydroxyl, yielding lactate, as NADH is oxidized to NAD+
.
Lactate, in addition to being an end-product of fermentation, serves as a
mobile form of nutrient energy, & possibly as a signal molecule in
mammalian organisms.
Cell membranes contain carrier proteins that facilitate transport of lactate.
Glycogen Glucose
Hexokinase or Glucokinase
Glucose-6-Pase
Glucose-1-P Glucose-6-P Glucose + Pi
Glycolysis
Pathway
Pyruvate
Glucose metabolism in liver.
Energy from glycolysis
ATP consumed 2 moles
ATP produced direct 4 moles
ATP indirect (NADH/H) 6 moles
Net ATPs = 10-2= 8 moles
If anaerobic glycolysis 2 moles
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Digestion glycolysis

  • 1. Digestion and Absorption of carbohydrates
  • 2.  CHO taken in diet are:  polysaccharides,  disaccharides  monosaccharides  These are supplied from external sources, hence called exogenous CHO  These may be  digestible or  indigestible Absorption and digestion of carbohydrates
  • 3. Digestion of CHO takes place in;  mouth stomach Intestine  Absorption takes place form small the intestine
  • 4. Digestion: Mouth At slightly acidic pH, salivary amyalse (ptyalin) acts on starch, which is converted into maltose and isomaltose The enzyme get inactivated in stomach 2(C6H10O5)n + nH2O nC12H22O11 starch maltose and isomaltose
  • 5. Stomach • HCl can cause hydrolysis of starch into maltose and isomaltose and that of maltose to glucose but the reaction is of little significance inside stomach Small intestine In the small intestine pancreatic amylase converts 87% starch to maltose and isomaltose and 13 % glucose (C6H10O5)n + nH2O nC6H12O6
  • 6. Disaccharides present in brush border of epithelial cells are lactose, sucrose, maltose and isomaltose Lactase, sucrase, maltase and isomaltase hydrolyze disaccharides into their components as: Sucrose into glucose and fructose Maltose into glucose Lactose into glucose and galactose
  • 7. Absorption Glucose (80-85%) and few disaccharides are absorbed from small intestine through capillaries of intestinal mucosa The order of ease of absorption of monosaccharides is galactose > glucose > fructose > mannose > pentose
  • 8. Mechanism of absorption Simple diffusion Active transport Factors upon which absorption of CHO depends are given as: Physical factors (how long food stays in intestine) Hormone thyroid hormones increase the rate of absorption hormones of adrenal cortex facilitate absorption
  • 9. Facts about absorption of monosaccharides Chemical nature of the most actively transported monosaccharides has following features in the choice of the carrier: Presence of 6 and more carbon atoms D-pyarnose structure Intact OH at carbon 2
  • 11. Glucose and glycogen are broken down in the body by a complex chain of reactions catalyzed by many enzymes There are many metabolic pathways by which glucose can be utilized in the body; the most important one is the Embden-Meyerhof pathway followed by citric acid cycle Glycolysis is of two types: anaerobic and aerobic
  • 12. Anaerobic glycolysis It takes place in cytosol (extra-mitochondrial) Glucose is broken into two molecules of pyruvic acid, which is then converted into lactic acid by utilizing NADH/H+ It can not continue indefinitely;  lactic acid lowers the pH to a level that is not suitable for cellular function On the other hand NADH/H+ becomes unavailable, if aerobic metabolism remains suspended for a long time
  • 13. Aerobic glycolysis In the presence of oxygen the pyruvic acid is formed in the same way as anaerobic glycolysis but it does not give rise to lactic acid Pyruvic acid is converted into acetyle-CoA which then enters in citric acid cycle Reactions of citric acid cycle occur in mitochondria
  • 14. Main Features The oldest of the Pathways Occurs in Soluble Phase of Cytoplasm (Cytosol) Anaerobic Phase Energy Generates ATP Produces Pyruvate/Lactate Produces Many Important Intermediates of other Pathways
  • 15. Relationship to Other Pathways TCA Cycle Gluconeogenesis (in Liver and Kidney) Hexose Monophosphate Shunt (HMP) Metabolism of other Sugars, e.g., Fructose and Galactose Metabolism of certain amino acids Lipid metabolism Glycoprotein Synthesis
  • 16. Transport of glucose into the cell Transportation of glucose is mediated by two types of systems that are given as follows: 1- Insulin-independent transport system Hepatocytes, erythrocytes and brain cells do not need insulin for the entry of glucose. Transport occurs by a protein, which is an oligomer (MW 200,000) containing 4 sub-units of equal size
  • 17. Transport of glucose into the cell 2- Insulin-dependent transport system It occurs in muscles and adipose tissue cells. The binding of insulin to the receptors enhances the transport of glucose into cell by causing o migration of glucose transport protein from microsomes to plasma membrane o and by increasing transport capacity of the transport proteins
  • 18. Glycolysis (Embden-Meyerhof pathway) Glycolysis takes place in the cytosol of the cells Glucose enters the glycolysis pathway by conversion to glucose-6-phosphate, which is initially an energy consuming step; energy input corresponding to one ATP H O OH H OHH OH CH2OPO3 2− H OH H 1 6 5 4 3 2 glucose-6-phosphate
  • 19. H O OH H OHH OH CH2OH H OH H H O OH H OHH OH CH2OPO3 2− H OH H 23 4 5 6 1 1 6 5 4 3 2 ATP ADP Mg2+ glucose glucose-6-phosphate Hexokinase 1. Hexokinase catalyzes: Glucose + ATP  glucose-6-P + ADP The reaction involves nucleophilic attack of C-6 hydroxyl of glucose by P of the terminal phosphate of ATP. ATP binds to the enzyme as a complex with Mg++
  • 20. Mg++ interacts with negatively charged phosphate oxygen atoms, providing charge compensation & promoting a favorable conformation of ATP at the active site of the Hexokinase enzyme N N N N NH2 O OHOH HH H CH2 H OPOPOP − O O O − O − O O O − adenine ribose ATP adenosine triphosphate
  • 21. The reaction catalyzed by Hexokinase is highly spontaneous A phosphoanhydride bond of ATP (~P) is cleaved The phosphate ester formed - glucose-6-phosphate - has a lower ∆G of hydrolysis H O OH H OHH OH CH2OH H OH H H O OH H OHH OH CH2OPO3 2− H OH H 23 4 5 6 1 1 6 5 4 3 2 ATP ADP Mg2+ glucose glucose-6-phosphate Hexokinase
  • 22. 2. Phosphoglucose Isomerase catalyzes: glucose-6-P (aldose)  fructose-6-P (ketose) H O OH H OHH OH CH2OPO3 2− H OH H 1 6 5 4 3 2 CH2OPO3 2− OH CH2OH H OH H H HO O 6 5 4 3 2 1 glucose-6-phosphate fructose-6-phosphate Phosphoglucose Isomerase
  • 23. 3. Phosphofructokinase catalyzes: fructose-6-P + ATP  fructose-1,6-bisP + ADP The Phosphofructokinase reaction is the rate-limiting step of Glycolysis. CH2OPO3 2− OH CH2OH H OH H H HO O 6 5 4 3 2 1 CH2OPO3 2− OH CH2OPO3 2− H OH H H HO O 6 5 4 3 2 1 ATP ADP Mg2+ fructose-6-phosphate fructose-1,6-bisphosphate Phosphofructokinase
  • 24. 4. Aldolase catalyzes: fructose-1,6-bisphosphate  dihydroxyacetone-P + glyceraldehyde-3-P The reaction is an aldol cleavage, the reverse of an aldol condensation. 6 5 4 3 2 1CH2OPO3 2− C C C C CH2OPO3 2− O HO H H OH H OH 3 2 1 CH2OPO3 2− C CH2OH O C C CH2OPO3 2− H O H OH+ 1 2 3 fructose-1,6- bisphosphate Aldolase dihydroxyacetone glyceraldehyde-3- phosphate phosphate Triosephosphate Isomerase
  • 25. 5. Triose Phosphate Isomerase catalyzes: dihydroxyacetone-P  glyceraldehyde-3-P 6 5 4 3 2 1CH2OPO3 2− C C C C CH2OPO3 2− O HO H H OH H OH 3 2 1 CH2OPO3 2− C CH2OH O C C CH2OPO3 2− H O H OH+ 1 2 3 fructose-1,6- bisphosphate Aldolase dihydroxyacetone glyceraldehyde-3- phosphate phosphate Triosephosphate Isomerase
  • 26. The ketose/aldose conversion involves acid/base catalysis, and is thought to proceed via an enediol intermediate, as with Phosphoglucose Isomerase. C C CH2OPO3 2− O C C CH2OPO3 2− H O H OH C C CH2OPO3 2− H OH OH H H OH H+ H+ H+ H+ dihydroxyacetone enediol glyceraldehyde- phosphate intermediate 3-phosphate Triosephosphate Isomerase
  • 27. C C CH2OPO3 2− H O H OH C C CH2OPO3 2− O OPO3 2− H OH + Pi + H+ NAD+ NADH 1 2 3 2 3 1 glyceraldehyde- 1,3-bisphospho- 3-phosphate glycerate Glyceraldehyde-3-phosphate Dehydrogenase 6. Glyceraldehyde-3-phosphate Dehydrogenase catalyzes: glyceraldehyde-3-P + NAD+ + Pi  1,3-bisphosphoglycerate + NADH + H+
  • 28. C C CH2OPO3 2− O OPO3 2− H OH C C CH2OPO3 2− O O− H OH ADP ATP 1 22 3 3 1 Mg2+ 1,3-bisphospho- 3-phosphoglycerate glycerate Phosphoglycerate Kinase 7. Phosphoglycerate Kinase catalyzes: 1,3-bisphosphoglycerate + ADP  3-phosphoglycerate + ATP
  • 29. C C CH2OH O O− H OPO3 2− 2 3 1 C C CH2OPO3 2− O O− H OH2 3 1 3-phosphoglycerate 2-phosphoglycerate Phosphoglycerate Mutase 8. Phosphoglycerate Mutase catalyzes: 3-phosphoglycerate  2-phosphoglycerate Phosphate is shifted from the OH on C3 to the OH on C2.
  • 30. 9. Enolase catalyzes: 2-phosphoglycerate  phosphoenolpyruvate + H2 O This dehydration reaction is Mg++ - dependent. C C CH2OH O O− H OPO3 2− C C CH2OH − O O− OPO3 2− C C CH2 O O− OPO3 2− OH− 2 3 1 2 3 1 H+ 2-phosphoglycerate enolate intermediate phosphoenolpyruvate Enolase
  • 31. 10. Pyruvate Kinase catalyzes: phosphoenolpyruvate + ADP  pyruvate + ATP C C CH3 O O− O2 3 1 ADP ATP C C CH2 O O− OPO3 2− 2 3 1 phosphoenolpyruvate pyruvate Pyruvate Kinase
  • 32. This phosphate transfer from PEP to ADP is spontaneous  PEP has a larger ∆G of phosphate hydrolysis than ATP  Removal of Pi from PEP yields an unstable enol, which spontaneously converts to the keto form of pyruvate C C CH3 O O− O2 3 1 ADP ATPC C CH2 O O− OPO3 2− 2 3 1 C C CH2 O O− OH2 3 1 phosphoenolpyruvate enolpyruvate pyruvate Pyruvate Kinase
  • 34. Glyceraldehyde-3-phosphate Dehydrogenase Phosphoglycerate Kinase Enolase Pyruvate Kinase glyceraldehyde-3-phosphate NAD+ + Pi NADH + H+ 1,3-bisphosphoglycerate ADP ATP 3-phosphoglycerate Phosphoglycerate Mutase 2-phosphoglycerate H2O phosphoenolpyruvate ADP ATP pyruvate
  • 35. C C CH3 O− O O C HC CH3 O− OH O NADH + H+ NAD+ Lactate Dehydrogenase pyruvate lactate E.g., Lactate Dehydrogenase catalyzes reduction of the keto in pyruvate to a hydroxyl, yielding lactate, as NADH is oxidized to NAD+ . Lactate, in addition to being an end-product of fermentation, serves as a mobile form of nutrient energy, & possibly as a signal molecule in mammalian organisms. Cell membranes contain carrier proteins that facilitate transport of lactate.
  • 36. Glycogen Glucose Hexokinase or Glucokinase Glucose-6-Pase Glucose-1-P Glucose-6-P Glucose + Pi Glycolysis Pathway Pyruvate Glucose metabolism in liver.
  • 37. Energy from glycolysis ATP consumed 2 moles ATP produced direct 4 moles ATP indirect (NADH/H) 6 moles Net ATPs = 10-2= 8 moles If anaerobic glycolysis 2 moles