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3 Oxidation of Hexoses
[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD
Dr. Reyes
31 July 2013
A. Fates of Glucose
1. Glucose transport
2. Hexokinase
3. HMP/Pentose phosphate pathway
4. Glycolysis
5. Lactate transport out of cell
6. Pyruvate decarboxylase
7. Krebs cycle
8. Glycogenesis
9. Glycogenolysis
10. Lipogenesis
11. Formation & release of VLDL
12. Gluconeogenesis
13. Hydrolysis of glucose-6-phosphate and release into the
blood
14. Glucuronide formation
I. Glycolysis
A. Fates of glucose
B. Balance
C. Three stages (10 steps) of glycolysis
II. Fate of pyruvate
A. Aerobic conditions
B. Anaerobic conditions
III. Regulation of glycolysis
A. Regulation of hexokinase IV
B. Regulation of phosphofructokinase I
C. Regulation of pyruvate kinase
IV. Mechanisms of Action
A. Of Hexokinase
B. Of Triosphosphate isomerase
C. Of GAP dehydrogenase
D. Of Phosphoglycerate kinase
E. Of Phosphoglycerate mutase
F. Inhibition of Enolase
G. Of Pyruvate kinase
V. Metabolism of other Hexoses
A. Metabolic fate of galactose
B. Metabolic fate of fructose
C. Metabolic fate of mannose
VI. Summary
VII. Competencies
VIII. Sample problems
Tissues
RBC
Brain
Muscle/Heart
Adipose
Liver parenchyma cells
OBJECTIVES
At the end of the lecture, the student should be able to:
1. Discuss the importance of glucose, galactose & fructose in energy
metabolism
2. Summarize the reactions and intermediates found in glycolysis
3. Enumerate the differences in the role of glycolysis in different cell
types
4. Discuss the three stages of glycolysis
5. Discuss the sources of ATP generation and utilization of glycolysis
6. Explain the relationship between lactate and pyruvate
7. Discuss the balance between NAD+ and NADH in the cell
8. Explain the primary controls of glycolysis
9. Explain the role of lactate dehydrogenase in the cell
Fates of Glucose
1, 2, 3, 4, 5
1, 2, 3, 4, 6, 7
1, 2, 3, 4, 5, 6, 7, 8, 9
1, 2, 3, 4, 6, 8, 9, 10
All
B. Balance
NAD+ and NADH
o Critical in the cytoplasm because this is where NAD is
required for glycolytic pathway to proceed
o Need for NAD+ in cytoplasm is closely related to levels
of pyruvate and lactate in the cytoplasm
o NAD is used by D-glyceraldehyde-3-phosphate
dehydrogenase
o Lactate dehydrogenase (anaerobic) will regenerate
levels of NADH in the cytoplasm
C. Three Stages (10 steps) of Glycolysis
References:
Devlin 7th ed.
2016 Trans
Stage
1. Priming
2. Splitting
I. GLYCOLYSIS
3. Oxidoreductionphosphorylation
Product
D-Fructose 1,6bisphosphate
D-Glyceraldehyde 3phosphate
L-Lactate (anaerobic
conditions)
Energy Yield
- 2 ATP
None
+ 4 ATP
Stage One:
o Trapping of glucose as glucose 6-phosphate within the
cytosol where all glycolytic enzymes are located
Stage Two:
o Reversible reaction, corresponding to an aldol cleavage
in one direction and an aldol condensation in the other
Stage Three:
o Payoff or harvest stage
o The overall reaction can be visualized as the coupling of
a very favorable exergonic reaction with an unfavorable
endergonic reaction
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Figure 1. Glycolysis
Go = -4.0 kcal/mol
Figure 2. Phosphorylation of Glucose
o Enzyme:
Hexokinase (hepatic cells) it has low Km for
glucose; thus once glucose enters the cell, it gets
phosphorylated
Glucokinase liver parenchymal cells and cells
of pancreas
o Phosphorylation of glucose by ATP is a
thermodynamically favorable, irreversible under cellular
conditions
2+
o ATP is coupled with Mg
Step 2: Glucose 6-phosphate Fructose 6-phosphate
Go = +4.0 kcal/mol
Go = -3.4 kcal/mol
Figure 4. Phosphorylation of F6P
Go = +5.7 kcal/mol
Go = +1.8 kcal/mol
Figure 5. Cleavage of F1,6BP and Interconversions of DHAP and GAP
Go = +1.06 kcal/mol
Go = +1.5 kcal/mol
Go = +0.44 kcal/mol
Figure 9. Dehydration of 2PG
Go = -6.1 kcal/mol
Substrate
Enzyme
Product
[E]
Phosphorylation
Hexokinase
G6PO4
- 1 ATP
Isomerization
Phosphohexose isomerase
F6PO4
None
F6PO4
Phosphorylation
Phosphofructokinase I
F1,6BP
- 1 ATP
F1,6BP
Cleavage of C3-C4
Aldolase
DHAP, G3P
None
Isomerization
GAP
None
Oxidoreduction
G3P dehydrogenase
1,3BPG
1 NADH (x2)
Phosphoglycerate kinase
3PG
1 ATP (x2)
3PG
Isomerization
Phosphoglycerate mutase
2PG
None
2PG
Dehydration
Enolase
PEP
None
Pyruvate kinase
Pyruvate
1 ATP (x2)
Oxidoreduction
Lactate dehydrogenase
Lactate
NAD+, H+
Glucose
G6PO4
DHAP
GAP, NAD
1,3BPG, ADP
PEP, ADP
*Pyruvate, NADH
Reaction
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2. Ethanol Fermentation
o NAD+ is regenerated by the fermentation of pyruvate to
ethanol and CO2
o Enzymes: pyruvate decarboxylase and alcohol
dehydrogenase
A. Aerobic Conditions
1. Transformation into Acetyl CoA
o Proceeds to Krebs cycle to form CO2 plus H2O;
eventually regenerates NAD+ and forms more ATP
through oxidative phosphorylation
o Enzyme: pyruvate dehydrogenase
3. Formation of Glucose
o Through gluconeogenesis
Enzyme
Hexokinase
I, II, III
PFK1
Pyruvate
kinase
Enhanced by
(+) allosteric effector
Decreased by
(-) allosteric effector
AMP, ADP
Glucose 6-phosphate
ATP, Citrate, H+
ATP, Alanine, Acetyl
CoA
B. Anaerobic Conditions
1. Lactate Fermentation
o Anaerobic glycolysis in contracting muscles
o Regeneration of NAD+ from NADH in states of hypoxia
o When lactate accumulates, an elevation of lactic acid in
the blood occurs (lactic acidosis)
o Enzyme: lactate Dehydrogenase
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- [5] 06 Baybay, Bascua, Basilan, Bataan, Batac, Bate, Baustista A, Bautista B, Bayani
+
Lys --- (CH2)4 NH3
+
Lys --- (CH2)4 NH3
-O
His
O O
II OC1
CH2
P
I
H-C2-O
ON
NH
I
O
H-C3- O
I
P
H O
O2,3-bishosphoglycerate
His
intermediate
-O
His
C1
(1)
O
CH2
I ..
II
H-C2-O-H
+
O-P N
NH
I
O I
H-C3- O
-O
I
P
Phosphoglycerate
Carboxy- H O
Omutase
late
anion of
3-phosphoglycerate
3-Phosphoglycerate
(2)
+
Lys --- (CH2)4 NH3
-O
CH2
O O
NH
II OC1
P
I
H-C2-O
OI
H-C3- OH
I
2-Phosphoglycerate
H
K+
Mg+2
-O
I
C
II
O
Phosphoryl O2
of ADP
OI
P - OO
II
I
C O
II
CH2
+
1
Phosphoenolpyruvate (PEP)
K+
A.
Metabolism of Galactose
Mg 2+
OOI
I
-O - P O - P - O - Adenosine
II
II
O
O-
ATP
ADP
Mg+2
-O
I
C
II
O
O
I
C
II
CH2
Enolpyruvate
2
-O
I
C
II
O
H+
O
II
C
I
CH3
Pyruvate
Inhibitors of Glycolysis
+2
+2
1. Sulfhydryl reagents (heavy metals like Hg and Ag ) and
alkylating agents like iodoacetate inhibit glyceraldehyde 3phosphate dehydrogenase via inhibition of formation of a
thiohemiacetal (formed via interaction between the sulfhydryl
[SH] group of a cysteine residue in the active site of the
enzyme glyceraldehyde 3-phosphate dehydrogenase and
the substrate glyceraldehyde 3-phosphate).
2. Hyperglycemia promotes overproduction of reactive O2
species that activate poly (ADP-ribose) polymerase (PARP)
polyation of cysteine residues of the active site of
glyceraldehyde 3-phosphate dehydrogenase nonformation of a thiohemiacetal.
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B. Non-classical galactosemia
o Deficiency of galactokinase
Accumulation of galactose in blood
(galactosemia) and urine (galactosuria)
o Galactose in diet
Accumulation of galactitol
Lactose intolerance
o Deficiency of -galactosidase (or lactase)
o Intestinal bacteria in lactase-deficient people hydrolyze
lactose
Causes osmotic diarrhea
o Common in premature infants, those which have
stomach surgery, mucosal damage after chronic bout of
diarrhea
o Milk and their products should be avoided
o Yoghurt can be tolerated in some because lactose has
been partially hydrolyzed via endogenous galactosidase of microorganism in yoghurt culture
o Commercial preparations of -galactosidase
Used to pretreat milk to reduce lactose content or
can be taken when milk products are ingested by
lactase-deficient individuals
B. Metabolism of Fructose
Via phosphoglucomutase
o Glucose-6-phosphate can then be converted to:
Pyruvate enters glycolysis
Free glucose via glycogenesis
Other information
Conversion of 1 molecule of galactose to 2 molecules of
pyruvate produces 2 molecules of ATP and 2 molecules of
NADH2 (same yield of glucose and fructose converted to
pyruvate)
Galactose metabolism
o Higher in infants than adults
o Blood level in systemic circulation is < 3 gm/dL
o No galactose lost in urine
Other sources of galactose
o Normal cell turnover
o Lysosomal degradation of glycolipids, glycoproteins and
glycosaminoglycans
I. Hexokinase Pathway
In muscles, adipose tissues, kidneys
Fructose metabolism in muscle differs little from glycolysis
Hexokinase has high Km
o Low fructose affinity
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o Glyceraldehyde DHAP
Enzyme: alcohol dehydrogenase
+
Initial reduction to glycerol by NAD
o DHAP Glycerol 3-phosphate
Enzyme: glycerol kinase
Phosphorylation of DHAP to glycerol 3-phosphate
in the presence of ATP
o Glycerol 3-phosphate DHAP
Enzyme: glycerol phosphate dehydrogenase
Reoxidation by NADH
Other information
Neither hexokinase nor phosphofructokinase can
phosphorylate fructose 1-phosphate at C6 to form the
glycolytic intermediate fructose 1,6-bisphosphate.
DHAP and GAP condense to form fructose 1,6-bisphosphate
that enters glycogenesis pathway
Glycerol 3-phosphate can enter TAG biosynthetic pathway
Metabolism of 1 molecule of fructose to 2 molecules of
pyruvate yields 2 molecules of ATP and 2 molecules of
NADH (same yield as conversion of glucose to pyruvate)
o Fate of fructose in parallel with glucose
Effect of bypassing PFK-1 reaction:
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VI. SUMMARY
Glucose is metabolized by all mammalian cells to form
ATP
Anaerobic glycolysis produces two molecules for each of
lactate and ATP from one molecule of glucose
Aerobic glycolysis produces two molecules each of NADH
and pyruvate
Glycolysis is regulated in three steps
+
NADH is formed from NAD during glycolysis
RedOx balance of the cell has to be maintained for furhter
cycles of glycolysis to continue
Mannose
o Minor diet component
o Unimportant energy source
o C2 epimer of glucose
Steps in Mannose Metabolism
Step 1: Mannose Mannose 6-phosphate
o Enzyme: hexokinase
o ATP as terminal phosphate donor
Step 2: Mannose 6-phosphate Fructose 6-phosphate
o Enzyme: phosphomannose isomerase
o Isomerization
o F6P enters glycolytic pathway
VII. COMPETENCIES
1. Given a well-fed person, identify the biochemical pathways or processes of carbohydrate metabolism involved.
2. Apply the biochemical concepts and principles that will help explain the development of diseases associated with defects in
carbohydrate metabolism.
3. Correlate the biochemical or molecular basis with the clinical manifestation of the disease, the findings on physical examination of the
patient and laboratory results.
VIII. SAMPLE PROBLEMS
1. Where does glycolysis take place in a cell?
2. In glycolytic steps one and three, what is the source of the phosphate groups that are added to glucose and fructose-6-phosphate,
respectively?
3. In step four of glycolysis, why is dihyroxyacetone phosphate (DHAP) immediately converted into glyceraldehyde-3-phosphate (GAP)
by the enzyme triosphosphate isomerase?
4. What two types of reactions does glyceraldehyde-3-phosphate dehydrogenase catalyze in the fifth step of glycolysis?
5. How many molecules of ATP are produced during the second stage of glycolysis?
6. What role does atomic magnesium play in step 6 of glycolysis involving the conversion of 1,3 bisphoglycerate to 3-phosphoglycerate?
7. What general name is given to an enzyme that catalyzes the transfer of a functional group from one position on a molecule to
another?
Answers:
1. Cytosol
2. ATP
3. DHAP is converted to GAP so that it can continue in glycolysis
4. Oxidation followed by phosphorylation
5. Four ATPs from steps 6 and 9
6. Magnesium functions to shield the negative charges
7. Mutase
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