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[BIOCHEMISTRY] 2.

3 Oxidation of Hexoses
[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD
Dr. Reyes

31 July 2013

06 Baybay, Bascua, Basilan, Bataan, Batac, Bate, Bautista A, Bautista B, Bayani


OUTLINE

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

Table 1. Fates of Glucose

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

Table 2. Stages of Glycolysis

Stage
1. Priming

Legend: Italicized quoted from the lecturer; bold emphasis, or


from references

2. Splitting

I. GLYCOLYSIS

3. Oxidoreductionphosphorylation

Embden-Meyerhof-Parnas (EMP) Pathway


Function:
o Glycolytic pathway in which anaerobic degradation of
glucose to lactate occurs with release of energy as ATP
o The major pathway for glucose and other hexoses
utilization to provide energy (ATP)
o Capable of producing 2 mol of ATP from 1 mol of
glucose in the absence of oxygen
o Emergency energy-yielding pathway
Site:
o Cytoplasm of all cells
o Extra-mitochondrial soluble fraction of the cell
Types:
o Aerobic uses oxygen; pyruvate is the end product
o Anaerobic uses no oxygen; lactate is the end product

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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C. Three stages (10 Steps of Glycolysis)

Figure 1. Glycolysis

STAGE 1: Priming Stage/Activation


Step 1: -D-Glucose + ATP Glucose 6-phosphate

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

Step 3: F6P + ATPFructose 1,6-bisphosphate + ADP

Go = -3.4 kcal/mol
Figure 4. Phosphorylation of F6P

o Enzyme: phosphofructokinase I (true rate limiting


enzyme or key regulatory enzyme of glycolysis)
o First committed step of glycolysis
o Irreversible phosphorylation of F6P to F1,6BP and uses
the second ATP needed to prime glucose
STAGE 2: Splitting/Cleaving Stage
Step 4: F1,6BP Dihydroxyacetone phosphate +
Glyceraldehyde 3-phosphate
o Enzyme: aldolase cleaves bond between C3 and C4
into two 3 carbon intermediates
C1-3 DHAP: Dihydroxyacetone phosphate
(ketone; need to be converted into aldehyde)
C4-6 GAP: Glyceraldehyde 3-Phosphate (true
substrate of the pathway)

Go = +5.7 kcal/mol

Figure 3. Isomerization of G6P

o Enzyme: phosphohexose isomerase


o Aldose-ketose isomerization: the reaction involves the
rearrangement of the carbon-oxygen bond to transform
the six-membered ring into a five-membered ring
o C6 is phosphorylated; C1 is susceptible to priming or
activation
o Reversible and is not subject to regulation

Go = +1.8 kcal/mol
Figure 5. Cleavage of F1,6BP and Interconversions of DHAP and GAP

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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


Step 5: DAHP Glyceraldehyde 3-phosphate
o Enzyme: Triose phosphate isomerase
o Ketone-aldehyde isomerization
o GAP is the only molecule that continues in the glycolytic
pathway
o 2 molecules of GAP are formed from each molecule
of glucose

Step 8: 3-phosphoglycerate 2-phosphoglycerate

Go = +1.06 kcal/mol

STAGE 3: Oxidoreduction reactions and ATP synthesis


+
Step 6: GAP 1, 3-bisphosphoglycerate + NADH + H

Figure 8. Isomerization of 3PG

o Enzyme: phosphoglycerate mutase


o Phosphoryl group transfer from C3 to C2
o 2,3-BPG is an obligatory intermediate at the active site

Go = +1.5 kcal/mol

Step 9: 2-phosphoglycerate Phosphoenolpyruvate

Figure 6. Oxidation and Phosphorylation of GAP

o Enzyme: glyceraldehyde 3-phosphate


dehydrogenase
o Oxidation of GAP is coupled to the reduction of NAD
First step when the cell starts to harvest energy
NADH will proceed to mitochondria for oxidative
phosphorylation

Go = +0.44 kcal/mol
Figure 9. Dehydration of 2PG

o Enzyme: enolase (lyase)


o A water molecule is removed to form
phosphoenolpyruvate which has a double bond between
C2 and C3
o This reaction generates a high energy phosphate from
one of markedly energy lower level

Step 7: 1,3BPG 3-phosphoglycerate + ATP


Go = +1.06 kcal/mol

Step 10: PEP + ADP + Mg2+ Pyruvate + ATP

Go = -6.1 kcal/mol

Figure 7. Transfer of High-energy Phosphate group

o Enzyme: phosphoglycerate kinase


o BPG has a mixed anhydride, a high-energy bond, at C1
high energy bond is hydrolyzed to a carboxylic acid
and the energy released is used to generate ATP from
ADP
o First site of ATP production in glycolysis all of the
invested ATP is recovered
o Bypassed in erythrocytes via Rappaport-Luebering
shunt

Figure 10. Formation of Pyruvate

o Enzyme: pyruvate kinase


o Second site of subtrate-level phosphorylation
o ADP has to be paired with magnesium ion for substrate
level phosphorylation to continue
o Irreversible reaction

Table 3. Glycolysis (summarized)

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

Triose phosphate isomerase

GAP

None

Oxidoreduction

G3P dehydrogenase

1,3BPG

1 NADH (x2)

Substrate level phosphorylation

Phosphoglycerate kinase

3PG

1 ATP (x2)

3PG

Isomerization

Phosphoglycerate mutase

2PG

None

2PG

Dehydration

Enolase

PEP

None

Substrate level phosphorylation

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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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


II. FATE OF PYRUVATE

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

Figure 14. Conversion of Pyruvate to Ethanol

o Occurs in some cells (like yeasts) and some anaerobic


organisms
o Pyruvate is converted to ethanol (excreted as a waste
product), releasing CO2 in the process
o Pathway has commercial roles in beer and bread
manufacture
o Mammals lack pyruvate decarboxylase; hence they
cannot produce ethanol from pyruvate
III. REGULATION OF GLYCOLYSIS
Regulatory enzymes in glycolysis (irreversible enzymes)
o Hexokinase or Glucokinase
o Phosphofructokinase I (PFK1)
o Pyruvate kinase

Figure 11. Transformation into Acetyl CoA

2. Carboxylation into a 4-C fragment oxaloacetate


o Intermediate of Krebs cycle and glucose synthesis

Table 4. Regulation of Glycolysis

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

AMP, ADP, F2,6BP

ATP, Citrate, H+
ATP, Alanine, Acetyl
CoA

AMP, ADP, F1,6BP

A. Regulation of Hexokinase IV or Glucokinase


o Not inhibited by G6P, unlike HK I, II, and III
o Predominant enzyme for glucose phosphorylation in the
liver (and -cells of the pancreas)
o Higher Km functions only when intracellular
concentration of glucose in the hepatocyte is elevated
o Has a high Vmax allowing liver to effectively remove
glucose from the blood
o Glucokinase levels are increased by carbohydrate-rich
diets and insulin
o At low levels of glucose, HK IV translocates into the
nucleus to bind with a regulatory protein glucokinase
inhibitory protein (GK-RP) to become metabolically
inactive
During hypoglycemia, the liver will not compete with
brain for glucose
o Migrates back to the cytoplasm when blood glucose
level returns to normal active
o Fructose 6-P helps retain the hexokinase IV-GKRP
bond thus F6P regulates HK IV.
o Note: the sugar specificity of HK-IV is similar to the
other hexokinase isoenzymes

Figure 12. Carboxylation of Pyruvate

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

B. Regulation of Phosphofructokinase I (PFK-1)


o In Devlin, named as 6-Phosphofructo-1-kinase
o Rate limiting enzyme of glycolysis
o Controls the rate of G6P entry into glycolysis in most
tissues
o Allosteric enzyme with 6 binding sites
2 for substrates: Mg-ATP and F6P
4 allosteric regulatory sites
Inhibitory site for Mg-ATP

Figure 13. Reduction of Pyruvate to Lactate

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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


Inhibitory site for citrate and other anions
Activation site for AMP
Activation site for Fructose 2, 6-bisphosphate (F2,6BP) and other bisphosphates

C. Regulation of Pyruvate kinase


o PK exists as tissue-specific isoenzymes
o The form present in brain and muscle: no allosteric sites
so PK does not contribute to the regulation of glycolysis
in these tissues
o Liver PK isoenzyme can be inhibited through
phosphorylation by the cAMP-dependent protein kinase
and by a number of allosteric factors which include:
o Activation by F1,6-bisP which ties rate of PK
to that of PFK-1
o Inhibition by ATP, which signifies high
energy levels

Figure 16. Mechanism of Triosphosphate isomerase

C. Mechanism of GAP dehydrogenase


1. Glyceraldehyde 3-phosphate reacts with the sulfhydryl group
(-SH) of cysteine residue of the enzyme glyceraldehyde 3phosphate dehydrogenase (large circle) to generate a
thiohemiacetal.
a. Thioester intermediate formation: ester group has 3 bonding
sites occupied after the bond from cysteine dissolves.
Remaining site will now easily accept the phosphoryl group
from orthosphosphate.

Reciprocal regulation between Glycolysis and


Gluconeogenesis
A. Glycolysis on; Gluconeogenesis off
B. Glycolysis off; Gluconeogenesis on
C. Avoidance of futile cycles

2. Followed by an internal oxidation-reduction reaction: NAD (


small circle) is reduced to NADH; thiohemiacetal is oxidized
to high-energy thioester.
3. Thioester reacts witth Pi to form the mixed anhydride and
regenerate the sulfhydryl group.
4. The mixed anhydride dissociates from the enzyme and
+
exogenous NAD replaces the bound NADH.

Otherwise a futile cycle would be present when both


metabolic pathways occur simultaneously in a given tissue.
For this reason, glycolysis regulation will be simultaneously
taken up with gluconeogenesis regulation.
IV. MECHANISM OF ACTION
A. Catalytic Mechanism of Hexokinase
1. Hexokinase catalyzes a phosphoryl group-transfer
reaction.
2. The C-6 hydroxyl O2 of glucose nucleophilically attacks the
2-.
-phosphorus of MgATP
3. MgADP is displaced and glucose 6-phosphate is
generated.

Figure 17. Mechanism of Glyceraldehyde 3-phosphate dehydrogenase

D. Mechanism of Phosphoglycerate kinase


+2
1. In cells, ADP or ATP is always complexed with Mg ions.
2. The terminal phosphoryl O2 of ADP nucleophilically attacks
the C1 phosphorus atom of 1,3-bisphosphoglycerate to form
+2
the reaction products 3-phosphoglycerate and Mg -ATP
complex.
Figure 15a. Mechanism of Hexokinase

Figure 18a. Mechanism of Phosphoglycerate kinase

Figure 15b. Mechanism of Hexokinase

B. Mechanism of Triosphosphate Isomerase


1. Triosphosphate isomerase catalyzes reversible
interconversion of DHAP and GAP
2. Transformation of DHAP into GAP one molecule of glucose
is converted into two molecules of GAP
Figure 18b. Phosphoglycerate bypass in Erythrocytes

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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


3. Arsenate prevents net synthesis of ATP without inhibiting
glycolysis.

E. Catalytic Mechanism of Phosphoglycerate mutase


1. A lysine residue at the active site of phosphoglycerate
mutase binds the carboxylate anion of 3-phosphoglycerate;
a histidine residue which is phosphorylated before the
substrate binds, donates its phosphoryl group to form 2,3bisphosphoglycerate intermediate.
2. Rephosphorylation of the enzyme with a phosphoryl group
from the C-3 position of the intermediate yields 2phosphoglycerate.

+
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

V. METABOLISM OF OTHER HEXOSES

3-Phosphoglycerate

(2)

+
Lys --- (CH2)4 NH3

1. A lysine residue of the active site


of phosphoglycerate mutase binds
the carboxylate anion of 3-phosphoglycerate; a histidine residue
which is phosphorylated before the
substrate binds, donates its phosphoryl group to form 2,3-bisphosphoglycerate intermediate.
2. Rephosphorylation of the enzyme with
a phosphoryl group from the C-3 of the
intermediate yields 2-phosphglycerate.

-O

Figure 21. Overview of Hexoses metabolism


O
II
-O-P +
N
I
-O

CH2

All are important sources of energy, like glucose


Initial phosphorylation is needed for them to be
metabolically useful
Entry to glycolytic pathway

O O
NH
II OC1
P
I
H-C2-O
OI
H-C3- OH
I
2-Phosphoglycerate
H

o Galactose via glucose-6-phosphate


o Mannose via fructose-6-phosphate
o Fructose
Via fructose-6-phosphate (in muscles, kidney, and
adipose tissues)
Via fructose 1,6-bisphosphate (in liver)

Figure 19. Phosphoglycerate mutase in isomerization of 3PG to 2PG

F. Mechanism of Action: Inhibition of Enolase


1. Fluoride is a potent inhibitor of Enolase
+2
2. Mg and Pi form an ionic complex with fluoride that inhibits
enolase by interfering with the binding of its substrate.

Thus, these hexoses behave like glucose also and undergo


the rest of glycolytic steps produce ATP, NADH,
pyruvate/lactate

G. Catalytic Mechanism of Pyruvate kinase


1. The phosphoryl oxygen of ADP nucleophilically attacks the
PEP phosphorus atom, thereby displacing the enol pyruvate
and forming ATP.
2. Enolpyruvate converts to pyruvate.
PEP phosphorus atom

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

1. The phosphoryl O2 of ADP


nucleophilically attacks the
PEP phosphorus atom, thereby
displacing the enol pyruvate and
forming ATP.
2. Enolpyruvate converts to
pyruvate .

Figure 22a. Metabolism of Galactose

Pyruvate

Figure 20. Mechanism of Pyruvate kinase

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.

Figure 22b. Synthesis of Glucose from Galactose

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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


Galactose C4 epimer of glucose
o 4 reactions required before galactose can be converted
to glycolytic intermediate glucose-6-phosphate

o Galactose conversion to galactitol via aldose


reductase in polyol pathway
Cataract formation (in hypergalactosemia than
hyperglycemia)
o Liver damage
Formation of cataracts, mental retardation, and
early death
o Children with galactosemia
Vomiting, diarrhea when drinking milk due to
enlarged liver and jaundice
o Special diet is needed (little galactose and lactose,
artificial milk made from casein or soybean
hydrosylate) to avoid severe effects

Steps in Galactose Metabolism


Step 1: Lactose Galactose Galactose 1-phosphate
o Lactose Galactose
Via intestinal lactase of -galactosidase
o Galactose Galactose 1-phosphate
Via galactokinase (with ATP as PO4 group donor
2+
and Mg as cofactor)
Galactose is phosphorylated at C1 to produce
galactose-1-phosphate
o Entry of glucose to liver is insulin independent (in
mammals)

B. Non-classical galactosemia
o Deficiency of galactokinase
Accumulation of galactose in blood
(galactosemia) and urine (galactosuria)
o Galactose in diet
Accumulation of galactitol

Step 2: Galactose 1-phosphate Glucose 1-phosphate +


UDP-galactose

o UDP-glucose Galactose 1-phosphate


Via galactose 1-phosphate uridyl transferase
transfers uridyl
o Galactose 1-phosphate UDP-galactose
Condensation of galactose from galactose 1phosphate with UDP of UDP-glucose
o Galactose 1-phosphate Glucose 1-phosphate
Condensation of phosphate group from galactose
1-phosphate with glucose of UDP-glucose

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

Step 3: UDP-galactose UDP-glucose


+
Via NAD -dependent UDP-galactose 4-epimerase
o UDP glucose Free glucose
Via glycogenesis in entering the glycogenesis
pathway
o UDP galactose can now be a donor of galactose units
in:
Lactose synthesis UDP galactose condenses with
glucose via lactose synthase to form lactose in
mammary glands
Synthesis of glycoproteins, glycolipids, and
glycosaminoglycans

B. Metabolism of Fructose

Step 4: Glucose 1-phosphate Glycolytic intermediate


Glucose 6-phosphate

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

Figure 23. Metabolism of Fructose

Fructose from dietary sucrose like honey, fruits, corn


syrup; from hydrolysis of sucrose or table sugar
o Enters pathway of intermediary metabolism via
phosphorylation via hexokinase pathway and
fructokinase pathway

Disorders of Galactose Metabolism


Galactosemia
A. Classical galactosemia
o Genetic absence of galactose 1-phosphate uridyl
transferase
Accumulation of galactose 1-phosphate in
tissues and galactose in blood and urine

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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[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


o Fructose is more rapidly glycolyzed or oxidized than
glucose
o More pyruvate (and acetyl CoA) is formed than is
required for ATP formation
Diet rich in fructose or sucrose leads to fatty liver
because of overproduction of pyruvate- precursor in
synthesis of fats and cholesterol- lipogenic effect of
fructose

o Less favored and much slower unless fructose


consumption is high
Steps in Hexokinase Pathway
Step 1: Fructose Fructose 6-phosphate
o Enzyme: hexokinase same hexokinase that
phosphorylates glucose to G6P
o Fructose phosphorylated to F6P in entry to muscle eclls
with ATP as terminal phosphate donor
o Only one reaction step prior to its entry to glycolysis
Therefore, it follows the rest of glycolytic steps

Polyol or Sorbitol Pathway

Step 2: F6P Fructose 1,6-bisphosphate


o Enzyme: phosphofructokinase I
o Irreversible step of glycolysis
Step 3: F1,6P DHAP + GAP
o Enzyme: aldolase A
II. Fructokinase Pathway
Major pathway (in liver and small intestines)
Fructokinase pathway more favored pathway and major
route for fructose phosphorylation
6 enzymes to convert fructose to glycolytic intermediates

Figure 24. Polyol/Sorbitol pathway

Glucose oxidized or reincorporated to glycogen in normal


conditions
Glucose fructose or vice versa (via sorbitol) without
phosphorylation in mammalian tissues (lens of eye, testes,
pancreas, brain)
Glucose reduced to sorbitol (glucitol) via NADPHdependent aldolase reductase
o High Km, low glucose affinity
o Glucose is usually metabolized by glycolysis
+
Sorbitol fructose via NAD dependent sorbitol
dehydrogenase (polyol dehydrogenase)
Present in seminal vesicles
o Sperm cells utilization of fructose, then switches to
glucose utilization when inside female reproductive tract
Increased concentration of glucose leads to increased
amounts of sorbitol in the said mammalian tissues
Less sorbitol dehydrogenase activity compared to aldose
reductase
o Sorbitol accumulate inside cell mebranes (relatively
impermeable to sorbitol)
o Increased osmolarity of cells
o Aggregation and precipitation of lens proteins
o Cataract in diabetics sorbitol accumulation in lens

Steps in Fructokinase Pathway


Step 1: Fructose Fructose 1-phosphate
o Enzyme: fructokinase present only in liver
o ATP as terminal phosphate donor
Step 2: Fructose 1-phosphate D-glyceraldehyde + DHAP

o Enzyme: aldolase B or fructose 1-phosphate


aldolase found only in liver
o Products: Trioses
o Rate-limiting step
Step 3: Glyceraldehyde Glyceraldehyde 3-phosphate (GAP)

o Enzyme: triokinase or glyceraldehyde kinase


o Direct phosphorylation by ATP
Alternate: Glyceraldehyde DHAP Glycerol 3-phosphate
DHAP

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

Disorders of Fructose Metabolism


Enzyme deficiencies
o Essential fructosuria
Rare; due to lack of hepatic fructokinase
No toxic metabolites of fructose occurs in liver
benign asymptomatic or benign condition
Fructosemia and fructosuria develops on fructose
consumption secondary to impaired fructose
utilization
Some fructose is phosphorylated by hexokinase in
non-hepatic tissues; some appear in urine

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:

o Hereditary fructose intolerance


Lack of aldolase B
Accumulation of fructose 1-PO2 in liver, kidney
damage and hypoglycemia from inhibition of
gluconeogenesis & glycogenolysis
Self-limiting; development of strong distaste for
anything sweet
Substrate overproduction
o Diet rich in fructose or glucose may lead to fatty liver
because of overproduction of pyruvate which is a

- [8] 06 Baybay, Bascua, Basilan, Bataan, Batac, Bate, Baustista A, Bautista B, Bayani

[BIOCHEMISTRY] 2.3 Oxidation of Hexoses Reyes, MD


Mannose via fructose-6-phosphate can also enter
mainstream of gluconeogenesis, glycogenesis, HMP and
uronic acid pathway (similar to glucose)

precursor in synthesis of fats and cholesterol via acetyl


coA
C. Mannose Metabolism

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

Figure 25. Mannose metabolism

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

Figure 26. Summary of entry of CHO from diet into glycolysis

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

- [9] 06 Baybay, Bascua, Basilan, Bataan, Batac, Bate, Baustista A, Bautista B, Bayani