BCH 312_Metabolism of Carbohydrates

(Dr. E.C. Ossai)

GLUCONEOGENESIS

The central role of glucose in metabolism arose early in evolution, and this sugar remains the
nearly universal fuel and building block in modern organisms, from microbes to humans. In
mammals, some tissues depend almost completely on glucose for their metabolic energy. For the
human brain and nervous system, as well as the erythrocytes, testes, renal medulla, and embryonic
tissues, glucose from the blood is the sole or major fuel source. The brain alone requires about 120
g of glucose each day—more than half of all the glucose stored as glycogen in muscle and liver.
However, the supply of glucose from these stores is not always sufficient; between meals and
during longer fasts, or after vigorous exercise, glycogen is depleted. For these times, organisms
need a method for synthesizing glucose from noncarbohydrate precursors through a pathway called
gluconeogenesis (“formation of new sugar”), which converts pyruvate and related three- and four-
carbon compounds to glucose.

Gluconeogenesis occurs in all animals, plants, fungi, and microorganisms. The reactions are
essentially the same in all tissues and all species. The important precursors of glucose in animals
are three-carbon compounds such as lactate, pyruvate, and glycerol, as well as certain amino acids
(Fig. 1). In mammals, gluconeogenesis takes place mainly in the liver, and to a lesser extent in
renal cortex. The glucose produced passes into the blood to supply other tissues. After vigorous
exercise, lactate produced by anaerobic glycolysis in skeletal muscle returns to the liver and is
converted to glucose, which moves back to muscle and is converted to glycogen—a circuit called
the Cori cycle. In plant seedlings, stored fats and proteins are converted, via paths that include
gluconeogenesis, to the disaccharide sucrose for transport throughout the developing plant.
Glucose and its derivatives are precursors for the synthesis of plant cell walls, nucleotides and
coenzymes, and a variety of other essential metabolites. In many microorganisms, gluconeogenesis
starts from simple organic compounds of two or three carbons, such as acetate, lactate, and
propionate, in their growth medium.

Gluconeogenesis and glycolysis are not identical pathways running in opposite directions,
although they do share several steps (Fig. 2); seven of the ten enzymatic reactions of
gluconeogenesis are the reverse of glycolytic reactions. However, three reactions of glycolysis are

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essentially irreversible in vivo and cannot be used in gluconeogenesis: the conversion of glucose to
glucose 6-phosphate by hexokinase, the phosphorylation of fructose 6-phosphate to fructose 1,6-

Figure 1: Carbohydrate synthesis from simple precursors

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bisphosphate by phosphofructokinase-1, and the conversion of phosphoenolpyruvate to pyruvate
by pyruvate kinase (Fig. 2). In cells, these three reactions are characterized by a large negative
free-energy change, ΔG, whereas other glycolytic reactions have a ΔG near 0.

Figure 2: Opposing pathways of glycolysis and gluconeogenesis

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In glycolysis, glucose is converted into pyruvate; in gluconeogenesis, pyruvate is converted into
glucose. However, gluconeogenesis is not a reversal of glycolysis. Several reactions must differ
because the equilibrium of glycolysis lies far on the side of pyruvate formation. The actual ∆G for
the formation of pyruvate from glucose is about -20 kcal mol-1 (-84 kJ mol-1) under typical cellular
conditions. Most of the decrease in free energy in glycolysis takes place in the three essentially
irreversible steps catalyzed by hexokinase, phosphofructokinase, and pyruvate kinase.

In gluconeogenesis, the following new steps bypass these virtually irreversible reactions of
glycolysis:

1. Phosphoenolpyruvate is formed from pyruvate by way of oxaloacetate through the action of

pyruvate carboxylase and phosphoenolpyruvate carboxykinase.

2. Fructose 6-phosphate is formed from fructose 1,6-bisphosphate by hydrolysis of the phosphate

ester at carbon 1. Fructose 1,6-bisphosphatase catalyzes this exergonic hydrolysis.

3. Glucose is formed by hydrolysis of glucose 6-phosphate in a reaction catalyzed by glucose 6-

phosphatase.

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GLYCOGENESIS

This involves intracellular synthesis of glycogen, i.e., taking up of excess glucose and converting
them to glycogen. Glycogenesis occurs only in the muscle and the liver. The first reaction in
glycogen synthesis involves the phosphorylation of glucose to glucose-6-phoshate (G.6.P), which
is a reaction catalyzed by glucokinase in the liver (hepatic cells) and hexokinase in the muscle
(peripheral tissues). G.6.P is converted to glucose-1-phosphate in a reaction catalyzed by
phosphoglucomutase. A unique reaction found in the next step involves the formation of UDP-
glucose by the action of G.1.P-uridyltransferase.

!"#$%&' + !"# → !. 6. ! + !"#

!. 6. ! → !. 1. !

!. 1. ! + !"# → !"# − !"#$%&' + !!"

The reaction generates an “activated” glucosyl residue, which can be used to build the glycogen
molecule. The formation of UDP-glucose is made energetically favourable and the reaction is
irreversible by subsequent hydrolysis of pyrophosphate by pyrophosphatase.

!!" + !! ! → 2!"

By the action of the enzyme, glycogen synthase, the C1 of the activated glucose of UDP-glucose
forms a glycosidic bond with a C4 of a terminal glucose residue of glycogen, liberating UDP
(uridine diphosphate). A pre-existing glycogen molecule or glycogen primer must be present to
initiate the reaction. The glycogen primer may in turn be formed on a protein primer known as
glycogenin.

The reducing end of glucose (C1) is always added to the non-reducing end of glycogen chain in a
1 → 4 linkage. Glycogen synthase will create chains of glucose molecules with !(1 → 4) linkages
but will not participate in the formation of !(1 → 6) linkages. Its action alone will produce
amylose, which is the straight chain polymer of glucose with !(1 → 4) linkages. Once an amylose
chain of at least 11 residues has been formed, a “branching” enzyme come into play. Its name is
amylo- ![1 → 4] → [1 → 6]-transglucosidase, which transfers a part of the 1 → 4 chain,
minimum length, 6-glucose residues to a neighbouring chain to form a 1 → 6 linkage, thus,
establishing a branch point in the molecule. The branches grow by further addition of 1 → 4
glucosyl unit and further branching. A new branch is introduced, at least, 4-glucosyl residues from
an adjacent branch point. This process synthesizes glycogen with multiple branch points.

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Notice that this enzyme is named for the reverse reaction; the same or another glycogen chain, thus creating a new
in the cell, the reaction proceeds in the direction of UDP- branch (Fig. 15–9). Further glucose residues may be
glucose formation, because pyrophosphate is rapidly added to the new branch by glycogen synthase. The
hydrolyzed by inorganic pyrophosphatase (Fig. 15–7). biological effect of branching is to make the glycogen
UDP-glucose is the immediate donor of glucose res- molecule more soluble and to increase the number of
idues in the reaction catalyzed by glycogen synthase, nonreducing ends. This increases the number of sites
which promotes the transfer of the glucose residue from accessible to!.!.!!!"#$%&
!!!"#!!! glycogen phosphorylase and glycogen syn-
!"#$%&'()"
UDP-glucose to a nonreducing end of a branched glyco- thase, both of!"#$%&'"#%'
!"#$%&#'()* which act only at nonreducing ends.
!"#$%&' + !"# !"# + !. 6. ! !. 1. ! !"# − !"#$%&'

6 CH
2OH
5
O
H H
H
4 1
OH H
HO O
3 2
H HO
O
"
O P O P O"
CH2OH CH2OH
O O CH2
Uracil O O
O H H H H
H H
UDP-glucose 4 1 4 1
H H OH H OH H
H H HO O O
OH OH H OH H OH
Nonreducing end of
a glycogen chain
glycogen
synthase with n residues
UDP (n # 4)

CH2OH CH2OH CH2OH

O O O
H H H H H H
New nonreducing H H H
4 1 4 1 4 1
end OH H OH H OH H
HO O O O
H OH H OH H OH
Elongated glycogen
with n ! 1 residues

FIGURE 15–8 Glycogen synthesis. A glycogen chain is elongated by glycogen synthase. The en-
zyme transfers the glucose residue of UDP-glucose to the nonreducing end of a glycogen branch
(see Fig. 7–15) to make a new (!1n4) linkage.

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DISORDERS OF CARBOHYDRATE METABOLISM

Disorders of carbohydrate metabolism are diseases associated with alteration in metabolism of

carbohydrate as a result of either deficiency of certain key enzymes or defect in mechanism.
Among the disorders of carbohydrate metabolism are diabetes mellitus, lactic acidosis, pyruvate
kinase deficiency and haemolytic anaemia, glycogen storage disease, fructose intolerance, glucose-
6-phosphate dehydrogenase deficiency and a host of others.

Fructose Intolerance Disease

Patients with hereditary fructose intolerance are deficient in the liver aldolase, which is an enzyme
responsible for splitting fructose-1-phosphate in the metabolism of fructose into dihydroxyacetone
phosphate and glyceraldehyde. Consumption of fructose by this patient results in the accumulation
of fructose-1-phosphate and in the depletion of inorganic phosphate (Pi) and ATP in the liver. The
reactions involved are those catalyzed by fructokinase and the enzymes of oxidative
phosphorylation.

Fructose + ATP = Fructose-1-phosphate + ADP

ADP + Pi + energy from electron transport chain = ATP

Pi + fructose = fructose-1-phosphate

Tying up of inorganic phosphate in the form of fructose-1-phosphate makes it impossible for liver
mitochondria to generate ATP by oxidative phosphorylation. The ATP level falls precipitously,
making it impossible for the liver to carry out its normal functions. Damage results to the cells in
large part because they are unable to maintain normal ion gradient by means of the ATP-dependent
cation pumps. The cells swell and eventually loose their internal content by osmotic lysis.

Pyruvate Kinase Deficiency and Haemolytic anaemia

Mature erythrocytes are absolutely dependent on glycolytic activity for ATP production. An ATP
is needed for the ion pump especially the Na+-K+-ATPase, which maintains the biconcave disc
shape of erythrocytes, which is the characteristic that helps erythrocytes slip through the capillaries
as they deliver oxygen to the tissues. Without ATP, the cells swell and lyse. Anaemia due to
excessive erythrocyte destruction is referred to as haemolytic anaemia. Pyruvate kinase deficiency
is rare but is by far the most common genetic defect of the glycolytic pathway known to cause
haemolytic anaemia. Most pyruvate kinase deficiency patients have 5 – 25% of normal red blood

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cell pyruvate level and flux through the glycolytic pathway is restricted severely, resulting in
markedly lower ATP level. The expected cross-over of the glycolytic intermediate is observed, i.e.,
those intermediates proximal to pyruvate kinase-catalyzed step accumulate whereas pyruvate and
lactate concentrations decrease.

Normal ATP levels are observed in reticulocytes of patients with this disease, this is because the
reticulocytes, which are younger red blood cells have mitochondria. Although deficiency in
pyruvate kinase, these immature cells have mitochondria and can generate ATP by oxidative
phosphorylation. Maturation of reticulocytes into red cells results in the loss of mitochondria and
complete dependent on glycolysis for ATP production. Since glycolysis is defective in this patient,
the mature cells are lost rapidly from the circulation. Anaemia results because the cells cannot be
replaced rapidly enough by erythropoiesis.