Carbohydrate Metabolism

Introduction and Digestion

Kinyi
Review of CHO Structure and Function

GLUCOSE
FRUCTOSE
 CHO DIGESTION
• Major dietary polysaccharides are of plant
(starch) and animal (glycogen) origin.
• Digestion starts in the mouth by salivary amylase
(pH of 6.4).
– Acts on dietary starch and glycogen in a random
manner,
– Hydrolyzing some α(1→4) glycosidic bonds.
– Cannot hydrolyze α(1→6) bonds
– Digestion results in a mixture of short, branched
oligosaccharides or dextrins.
 CHO DIGESTION
• Stomach-HCL inhibits salivary amylase- no
digestion.
• Small intestine-pancreatic α-amylase.
– Products -oligosaccharides, disaccharides and
monosaccharides.
 CHO DIGESTION
• Mucosal lining of the upper jejunum-
disaccharidases and oligosaccharidases.
– Isomaltase cleaves the α(1→6) bond in isomaltose
– maltase cleaves maltose
– sucrase cleaves sucrose producing glucose and
fructose
– lactase (β-galactosidase) cleaves lactose producing
galactose and glucose.
 Absorption
• Site: the duodenum and upper jejunum
• Insulin is not required
• Via an an active, energy-requiring process that
requires a concurrent uptake of sodium ions.
• Fructose uptake requires a sodium-independent
monosaccharide transporter (GLUT-5) for its
absorption.
• All three monosaccharides are transported from
the intestinal mucosal cell into the portal
circulation by yet another transporter, GLUT-2.
Absorption
 Digestive enzyme deficiencies
• Hereditary deficiencies of individual disaccharidases
(infants and children)
• Can also be caused by intestinal diseases, malnutrition,
or drugs that injure the mucosa of the small intestine.
• For example, brush border enzymes are rapidly lost in
normal individuals with severe diarrhea, causing a
temporary, acquired enzyme deficiency. Thus, patients
suffering or recovering from such a disorder cannot
drink or eat significant amounts of dairy products or
sucrose without exacerbating the diarrhea.
 Lactose intolerance
• The mechanism by which this age-dependent loss
of the enzyme occurs is not clear, but it is
determined genetically and represents a
reduction in the amount of enzyme protein
rather than a modified inactive enzyme.
• Treatment for this disorder is to reduce
consumption of milk while eating yogurts and
cheeses, as well as green vegetables such as
broccoli, to ensure adequate calcium intake; to
use lactase-treated products; or to take lactase in
pill form prior to eating.
 Entry of glucose into cells
• Insulin-independent transport system of
glucose :
• This is a carrier mediated uptake of glucose
which is not dependent on the hormone
insulin
• T his is operative in:
– hepatocytes
– Erythrocytes
– brain
 Entry of glucose into cells
• Insulin-dependent transport system :
– This occurs in muscle and adipose tissue.
– Glucose transporters : In recent years, at least six
glucose transporters(GLUT-l to GLUT-5 and GLUT-7) in
the cell membranes have been identified.
– They exhibit tissue specificity. For instance, GLUT-I is
abundant in erythrocytes whereas GLUT-4 is abundant
in skeletal muscle and adipose tissue.
– Insulin increases the number and promotes the
activity of GLUT-4 in skeletal muscle and adipose
tissue.
– In type 2 diabetes mellitus, insulin resistance is
observed in these tissues. This is due to the reduction
in the quantity of GLUT-4 in insulin deficiency.
GLYCOLYSIS
 Glycolysis
• Pathway employed by all tissues for the
breakdown of glucose
• Main purpose is to provide energy (in the form of
ATP) and intermediates for other metabolic
pathways.
• Pyruvate is the end product of glycolysis in cells
with mitochondria and an adequate supply of
oxygen (aerobic)
• Alternatively, pyruvate is reduced to lactate as
NADH is oxidized to NAD+ (anaerobic glycolysis)
 Reactions of glycolysis
• In glycolysis, a molecule of glucose is
degraded in a series of enzyme catalysed
reactions to yield two molecules of pyruvate.
• The free energy released from glucose is
conserved in the form of ATP and NADH.
• Glycolysis has two phases;
– The preparatory phase
– Pay off phase
 The preparatory phase
• Here the energy of ATP is invested raising the free
energy content of the intermediates.
• The carbon chains of all the metabolized hexoses are
converted into a common product, glyceraldehyde-3-
phosphate.
 Pay off phase
• In this phase energy is conserved by coupled
phosphorylation of four molecules of ADP to ATP.
• The net yield is two molecules of ATP per molecule of
glucose used, because two molecules of ATP were
invested in the preparatory phase.
• Energy is also conserved in the pay off phase in the
formation of two molecules of NADH per mole of
glucose.
 Glycolysis
• Three reactions are worthy to note in
glycolysis:
– Degradation of carbon Skelton of glucose to yield
pyrurate
– Phosphorylation of ADP to ATP by high energy
phosphate compounds formed during glycolysis.
– Transfer of hydride ion to NAD+ forming NADH
 Poisons of Glycolysis
• Arsenate which inhibits formation of 1,3-
phosphoglycerate.
• Iodoacetate which inhibits the enzyme
aldolase.
• Fluoride which leads to the accumulation of
phosphoglycerate.
Summary of Glycolysis
Glucose + 2NAD+ + 2ADP + 2Pi → 2
pyruvate + 2NADH + 2H+ + 2ATP + 2H2O
Regulation of Glycolysis
IRREVERSIBLE STEPS OF GLYCOLYSIS
 Hexokinase reaction
• Mammals have two isozymes of the enzyme
hexokinase that catalyze the phosphorylation of
glucose to glucose 6-phosphate
• Hexokinase is inhibited by the reaction product,
glucose 6-phosphate, which accumulates when
further metabolism of this hexose phosphate is
reduced
• Glucokinase activity is not allosterically inhibited
by glucose 6-phosphate as are the other
hexokinases, but rather is indirectly inhibited by
fructose 6-phosphate, and is stimulated indirectly
by glucose.
Hexokinase reaction
 Phosphofructokinase-1
• PFK-1 is rate limiting enzyme and primary site
of regulation
– Allosterically stimulated by AMP e.g. High
glycolysis during exercise
– Allosterically inhibited by:
• ATP e.g. High energy, resting or low
exercise
• Citrate: Build up from Krebs’ cycle
 Phosphofructokinase-1
– Fructose 2,6-bisphosphate is the most potent activator
of PFK-1, and is able to activate the enzyme even when
ATP levels are high.
• Fructose 2,6-bisphosphate is formed by phosphofructokinase-2
(PFK-2), an enzyme different than PFK-1.
• The reciprocal actions of fructose 2,6-bisphosphate on glycolysis
(activation) and gluconeogenesis (inhibition) ensure that both
pathways are not fully active at the same time, preventing a
futile cycle in which glucose would be converted to pyruvate
followed by resynthesis of glucose from pyruvate.
Phosphofructokinase-1
 Pyruvate kinase
• is activated by fructose 1,6-bisphosphate, the
product of the phosphofructokinase reaction.
• This feed-forward regulation has the effect of
linking the two kinase activities: increased
phosphofructokinase activity results in
elevated levels of fructose 1,6-bisphosphate,
which activates pyruvate kinase.
 Hormonal Regulation
• The regulation of glycolysis by allosteric
activation or inhibition, rate-limiting enzymes,
is short term (minutes or hours).
• Hormonal effects are slower, and often more
profound and are mainly on the amount of
enzyme protein synthesized. These effects can
result in ten-fold to twenty-fold increases in
enzyme activity that typically occurs over
hours to days.
 Hormonal Regulation
• Regular consumption of meals rich in
carbohydrate or administration of insulin initiates
an increase in the amount of glucokinase,
phosphofructokinase, and pyruvate kinase in
liver.
• Conversely, synthesis of glucokinase,
phosphofructokinase, and pyruvate kinase are
decreased when plasma glucagon is high and
insulin is low, for example, as seen in fasting or
diabetes.
 Pasteur effect in glycolysis
• The rate and the total amount of glucose
consumed are many times greater under
anaerobic than aerobic conditions
• The ATP yield from glycolysis under anaerobic
conditions (2 ATP per molecule of glucose) is
much smaller than that from the complete
oxidation of glucose to CO2 under aerobic .
• About 15 times as much glucose must
therefore be consumed anaerobically as
aerobically to yield the same amount of ATP.
Glucose metabolism in Cancer cells
• Glucose uptake and glycolysis proceed about ten times
faster in most solid tumors
• Tumor cells commonly experience hypoxia causing cancer
cells more than 100 to 200 μm from the nearest capillaries
depend on anaerobic glycolysis for much of their ATP
production.
• They take up more glucose than normal cells, converting it
to pyruvate and then to lactate as they recycle NADH.
• The high glycolytic rate may also result in part from smaller
numbers of mitochondria in tumor cells; less ATP made by
respiration-linked phosphorylation in mitochondria means
more ATP is needed from glycolysis.
Glucose metabolism in Cancer cells
• In addition, some tumor cells overproduce
several glycolytic enzymes, including an isozyme
of hexokinase that associates with the cytosolic
face of the mitochondrial inner membrane and is
insensitive to feedback inhibition by glucose 6-
phosphate.
• This enzyme may monopolize the ATP produced
in mitochondria, using it to convert glucose to
glucose 6-phosphate and committing the cell to
continued glycolysis.
Glucose metabolism in Cancer cells
• The hypoxia-inducible transcription factor
(HIF-1) is a protein that acts at the level of
mRNA synthesis to stimulate the synthesis of
at least eight of the glycolytic enzymes.
• This gives the tumor cell the capacity to
survive anaerobic conditions until the supply
of blood vessels has caught up with tumor
growth.
FEEDER PATHWAYS OF GLYCOLYSIS
• Many carbohydrates besides glucose meet
their catabolic fate in glycolysis, after being
transformed into one of the glycolytic
intermediates.
• Glycogen in animal tissues is mobilized for use
by the enzyme glycogen phosphorylase,
generating glucose-1-phosphate.
FEEDER PATHWAYS OF GLYCOLYSIS
• D-Fructose present in free form in many fruits.

This is the major pathway of fructose entry into

glycolysis in the muscles and kidney.
 FEEDER PATHWAYS OF GLYCOLYSIS
• In liver however, fructose enters by a different pathway.
• Galactose the product of hydrolysis
of lactose (milk sugar).
• In the liver it is phosphorylated by
galacto kinase
• Galactose-1-phosphate is then
converted to glucose-1-phosphate
through a set of reactions involving
Uridine diphospate (UDP).
• Defects in enzymes vary from
development of cataracts in infants,
to poor growth in children, speech
abnormality, mental deficiency and
liver damage depending on the
deficient enzyme.
FEEDER PATHWAYS OF GLYCOLYSIS
Fates of Pyruvate
 Conversion to Acetyl-CoA
• In aerobic organisms, pyruvate is converted to
acetyl CoA.
• Acetyl CoA is then entered into the citric acid
cycle.
Pyruvate + CoA + NAD -> AcetylCoA + CO2 + NADH
Enzyme: Pyruvate Dehydrogenase Complex (PDH)
• Multi-enzyme complex
– Three enzymes
– 5 co-enzymes
– Allows for efficient direct transfer of product from one
enzyme to the next
 Pyruvate dehydrogenase complex
• The PDH complex is composed of multiple
copies of three enzymes:
– Pyruvate dehydrogenase, E1 (with its bound
cofactorTPP);
– Dihydrolipoyl transacetylase, E2 (with its
covalently bound lipoyl group);
– Dihydrolipoyl dehydrogenase, E3 (with its
cofactors FAD and NAD).
 Pyruvate dehydrogenase complex
• E1 catalyzes first the decarboxylation of
pyruvate, producing hydroxyethyl-TPP, and
then the oxidation of the hydroxyethyl group
to an acetyl group.
• E2 catalyzes the transfer of the acetyl group to
coenzyme A, forming acetyl-CoA.
• E3 catalyzes the regeneration of the disulfide
(oxidized) form of lipoate; electrons pass first
to FAD, then to NAD .
Conversion to Acetyl-CoA
 Reduction of pyruvate to lactate
• This occurs in vigorously contracting muscle
functioning under low oxygen conditions
(hypoxia).
• This is known as Anaerobic Glycolysis. NADH
cannot be reoxidised to NAD+, but NAD+ is
required as an electron acceptor for further
oxidation of pyruvate in certain tissues e.g.
retina and erythrocytes convert glucose to
lactate even under aerobic conditions.
Reduction of pyruvate to lactate
 Reduction of pyruvate to lactate
• The lactate formed by active skeletal muscles
(or by RBCs) can be recycled.
• It is carried in blood to the liver where it is
converted to glucose during recovery from
strenuous muscle activity.
• The cycle of reactions that include glucose
conversion from lactate in muscle and lactate
conversion to glucose in the liver are called
the cori cycle.
THE CORI CYCLE
 Formation of ethanol
(ethanol/alcohol fermentation)
• It occurs in some plant tissues, invertebrates
and brewer’s yeast.
• Alcohol dehydrogenase is present in many
organisms, including humans where it
catalyzes the oxidation of ethanol to
acetaldehyde
 Formation of ethanol
(ethanol/alcohol fermentation)