Carbohydrate Metabolism

Glycolysis
Assist. Prof. Derya CANSIZ
İstanbul Medipol University
School of Medicine
Biochemistry Deparment
Content
• Structure
• Functions
• Metabolism
✓ Glycolysis
✓ Glycogenesis
✓ Gluconeogenesis
✓ Glycogenolysis
✓ TCA
✓ Pentose Phosphate Pathway
✓ Uronic Acid Pathway
Introduction…..

• Disaccharides, oligosaccharides, and polysaccharides

consist of monosaccharides linked by glycosidic bonds.
• Compounds with the same chemical formula but
different structures are called isomers.
• Two monosaccharide isomers differing in configuration
around one specific carbon atom (not the carbonyl
carbon) are defined as epimers.
• Monosaccharides (simple sugars) can be
classified according to the number of carbon
atoms they contain.
• Carbohydrates with an aldehyde as their carbonyl
group are called aldoses, whereas those with a
keto as their carbonyl group are called ketoses
• glyceraldehyde is an aldose, whereas
dihydroxyacetone is a ketose.
• Monosaccharides can be linked by glycosidic bonds to
create larger structures
• Disaccharides contain two monosaccharide units,
Important disaccharides include
-Lactose (galactose +glucose)
-Sucrose (glucose + fructose)
-Maltose (glucose + glucose)
• Oligosaccharides contain three to ten monosaccharide
units
• Polysaccharides contain more than ten monosaccharide
units and can be hundreds of sugar units in length.
• Important polysaccharides include branched glycogen
(from animal sources) and starch (plant sources)
A glycosidic bond between two hexoses
producing a disaccharide.
Enantiomers
• A special type of isomerism is found in
the pairs of structures that are mirror
images of each other.
• These mirror images are called
enantiomers, and the two members of the
pair are designated as a D- and an L- D and L forms of alanine are mirror images
sugar.
• The vast majority of the sugars in
humans are D-isomers.
• In the D-isomeric form, the –OH group
on the asymmetric carbon (a carbon
linked to four different atoms or groups)
farthest from the carbonyl carbon is on
the right, whereas in the L-isomer, it is
on the left.
Anomers

• Creation of an anomeric carbon (the former carbonyl carbon) generates a new pair of isomers, the
α and β configurations of the sugar (for example, α-D-glucopyranose and β-D-glucopyranose).
• Reducing sugars: If the hydroxyl group on the anomeric carbon of a cyclized sugar is not linked
to another compound by a glycosidic bond, the ring can open.
• The sugar can act as a reducing agent and is termed a reducing sugar. Such sugars can react with
chromogenic agents (for example, the Benedict reagent) causing the reagent to be reduced and
colored as the aldehyde group of the acyclic sugar is oxidized to a carboxyl group.
• All monosaccharides, but not all disaccharides, are reducing sugars.

Fehling or the Benedict solution

• Colorimetric test can detect a reducing sugar in urine. A positive result is indicative of an
underlying pathology (because sugars are not normally present in urine).
What is the importance of Glucose?
Energy Pyruvate Lactate
Connective Blood Group Substances
Tissue
Molecules
Cholesterol and Fatty Acid
Synthesis
Glycolipid Glucose
glycoprotein
Antioxidant effect

Nucleic acid Glycogen

 DIETARY CARBOHYDRATE DIGESTION

• The principal sites of dietary carbohydrate digestion are

the mouth and intestinal lumen.
• This digestion is rapid and is catalyzed by enzymes
known as glycoside hydrolases (glycosidases) that
hydrolyze glycosidic bonds.
• The final products of carbohydrate digestion are the Hydrolysis of a glycosidic bond
monosaccharides glucose, galactose, and fructose that are
absorbed by cells (enterocytes) of the small intestine.
Digestion of Dietary Carbohydrates

• Salivary α-amylase initiates digestion of dietary polysaccharides (for example, starch or

glycogen), producing oligosaccharides.
• Pancreatic α-amylase continues the process.
• The final digestive processes occur at the mucosal lining of the small intestine.
A. Salivary α-amylase

• The major dietary polysaccharides are of plant (starch, composed of

amylose and amylopectin) and animal (glycogen) origin.
• During mastication (chewing), salivary α-amylase acts briefly on
dietary starch and glycogen, hydrolyzing random α(1→4) bonds.
• Carbohydrate digestion halts temporarily in the stomach, because the
high acidity inactivates salivary α-amylase.

B. Pancreatic α-amylase
• When the acidic stomach contents reach the small intestine, they are
neutralized by bicarbonate secreted by the pancreas, and pancreatic
α-amylase continues the process of digestion.

C. Intestinal disaccharidases
• The final digestive processes occur primarily at the mucosal lining of the
duodenum and upper jejunum and include the action of several
disaccharidases.

Digestion of carbohydrates
D. Intestinal absorption of monosaccharides
• The upper jejunum absorbs the bulk of the monosaccharide
products of digestion.
• However, different sugars have different mechanisms of absorption.
• Galactose and glucose are taken into enterocytes by secondary
active transport that requires a concurrent uptake (symport) of
sodium (Na+) ions.
• The transport protein is the sodium-dependent glucose cotransporter
1 (SGLT-1).
• Sugar transport is driven by the Na+ gradient created by the Na+-
potassium (K+) ATPase that moves Na+ out of the enterocyte and
K+ in
• Fructose absorption utilizes an energy- and Na+-independent
monosaccharide transporter (GLUT-5).
• All three monosaccharides are transported from the enterocytes into
the portal circulation by yet another transporter, GLUT-2.
Absorption by enterocytes of the
monosaccharide products of
carbohydrate digestion.
E. Abnormal degradation of disaccharides

• Lactose intolerance: Over 60% of the world’s adults are

lactose intolerant
• The age-dependent loss of lactase activity starting at
approximately age 2 years represents a reduction in the
amount of enzyme produced.
• Treatment for this disorder is to reduce consumption of
milk; eat yogurts and some cheeses (bacterial action and
aging process decrease lactose content) as well as green
vegetables, such as broccoli, to ensure adequate calcium
intake; use lactase-treated products; or take lactase in pill
form prior to eating.
 If energy is not required..

If energy is required..

16
GLYCOLYSIS
• The glycolytic pathway is used by all tissues for the oxidation of glucose to provide energy (as ATP) and
intermediates for other metabolic pathways.
• Glycolysis is at the hub of carbohydrate metabolism because virtually all sugars, whether arising from
the diet or from catabolic reactions in the body, can ultimately be converted to glucose (Fig. A)
• Pyruvate is the end product of glycolysis in cells with mitochondria and an adequate supply of O2.
• This series of ten reactions is called aerobic glycolysis because O2 is required to reoxidize the
NADH formed during the oxidation of glyceraldehyde 3-phosphate (Fig. B)
• Aerobic glycolysis sets the stage for the oxidative decarboxylation of pyruvate to acetyl CoA, a major fuel of
the TCA cycle.
• Alternatively, pyruvate is reduced to lactate as NADH is oxidized to NAD+ (Fig. C).
• This conversion of glucose to lactate is called anaerobic glycolysis because it can occur without the
participation of O2.
• Anaerobic glycolysis allows the production of ATP in tissues that lack mitochondria (for example, red blood
cells and parts of the eye) or in cells deprived of sufficient O2 (hypoxia).
 GLUCOSE TRANSPORT INTO CELLS
• Glucose cannot diffuse directly into cells but enters by one of two transport systems:
1. Sodium (Na+)-independent transport system (dependent cotransport system)
• Extracellular glucose binds to the transporter, which then alters its conformation,
transporting glucose across the cell membrane via facilitated diffusion.
Tissue specificity:
GLUT-3 is the primary isoform in neurons.
GLUT-1 is abundant in RBC and the blood–brain barrier but is low in adult muscle,
GLUT-4 is abundant in muscle and adipose tissue.
The number of GLUT-4 transporters active in these tissues is increased by insulin.
GLUT-2 is abundant in the liver, kidneys, and pancreatic β cells. The
other GLUT isoforms also have tissue-specific distributions.
Specialized functions:
GLUT-1, GLUT-3, GLUT-4 involved in glucose uptake from blood to tissue.
GLUT-2, in the liver and kidneys, can either transport glucose into these cells when
blood glucose levels are high or transport glucose from these cells when blood glucose
levels are low (for example, during fasting).
GLUT-5 primary transporter for fructose (not glucose) in the small intestine and testes.
2. ATP-independent transport system
• This energy-requiring process transports glucose
against (up) its concentration gradient (that is, from
low extracellular concentrations to higher
intracellular concentrations) as Na+ is transported
down its electrochemical gradient.
• The gradient is created by the Na+-potassium (K+)
ATPase. Because this secondary active transport
process requires the concurrent uptake (symport) of
Na+, the transporter is a sodium-dependent glucose
cotransporter (SGLT).
• This type of cotransport occurs in the epithelial cells
of the intestine and renal tubules
• Glucose released into the blood after absorption
rises up to the renal threshold of 180 mg/dL.
Once this limit is exceeded, glucose begins to
appear in the urine.
• In normal people, 2 hours after absorption, blood
sugar returns to fasting value because it is carried
to all cells by the effect of insulin and other
mechanisms.

The renal threshold for glucose (RTg) corresponds

to a blood glucose level of ~180 mg/dL
GLYCOLYSIS REACTIONS

• Rate-limited Enzymes
1. Hexokinase
2. Phosphofructokinase
3. Pyruvate kinase
1. Hexokinase/Glucokinase is rate limiting enzyme
• Glucokinase is hexokinase IV isozyme
• Predominant enzyme responsible for glucose
phosphorylation.
• In β cells, glucokinase functions as a glucose sensor,
determining the threshold for insulin secretion.
• Hexokinase IV also serves as a glucose sensor in
hypothalamic neurons, playing a key role in the adrenergic
response to hypoglycemia.
• In the liver, the enzyme facilitates glucose phosphorylation
during hyperglycemia.
2. Phosphofructokinase-1 (PFK-1) is the most important
control point and the rate-limiting and committed step of
glycolysis.
-Regulation by intracellular energy levels:
• PFK-1 is controlled by the available concentrations of the
substrates ATP and fructose 6-phosphate as well as by other
regulatory molecules.
-Regulation by fructose 2,6-bisphosphate:
• 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.
• It is formed from fructose 6-phosphate by PFK-2.
Regulation by fructose 2,6-bisphosphate:

a. During the well-fed state: Decreased levels of glucagon and elevated levels of insulin cause an
increase in hepatic fructose 2,6-bisphosphate (PFK-2 is dephosphorylated) and, thus, in the rate of
glycolysis.
• Therefore, fructose 2,6-bisphosphate acts as an intracellular signal of glucose abundance.
b. During fasting: By contrast, the elevated levels of glucagon and low levels of insulin that occur
during fasting cause a decrease in hepatic fructose 2,6-bisphosphate (PFK-2 is phosphorylated).
• This results in inhibition of glycolysis and activation of gluconeogenesis.
 1,3-Bisphosphoglycerate synthesis:
Arsenic poisoning

Energy-generating phase:
2,3-Bisphosphoglycerate synthesis in RBC:

• Some of the 1,3-BPG is converted to 2,3-BPG by the action of

bisphosphoglycerate mutase.
• 2,3-BPG, which is found in only trace amounts in most cells,
is present at high concentration in RBC and serves to increase
Fluoride inhibits enolase, and O2 delivery.
water fluoridation reduces lactate • 2,3-BPG is hydrolyzed by a phosphatase to 3-
production by mouth bacteria, phosphoglycerate, which is also an intermediate in glycolysis.
decreasing dental caries • In the RBC, glycolysis is modified by inclusion of these shunt
• reactions.
The conversion of PEP to pyruvate,
catalyzed by pyruvate kinase, is the
third irreversible reaction of glycolysis.
 Pyruvate Kinase Regulation
1. Feedforward regulation:
• PK is activated by fructose 1,6-bisphosphate, the product of the PFK-1
reaction.
• This feedforward (instead of the more usual feedback) regulation has the
effect of linking the two kinase activities: increased PFK-1 activity
results in elevated levels of fructose 1,6-bisphosphate, which activates
PK.
2. Covalent regulation in the liver:
• Phosphorylation by cAMP-dependent PKA leads to inactivation of the
hepatic isozyme of PK.
• When blood glucose levels are low, elevated glucagon increases the
intracellular level of cAMP, which causes the phosphorylation and
inactivation of PK in the liver only.
• Therefore, PEP is unable to continue in glycolysis and, instead, enters
the gluconeogenesis pathway.
• This partly explains the observed inhibition of hepatic glycolysis and
stimulation of gluconeogenesis by glucagon.
• Dephosphorylation of PK by a phosphatase results in reactivation of the
enzyme.
3. Pyruvate kinase deficiency:
• Because mature RBC lack mitochondria, they are
completely dependent on glycolysis for ATP
production.
• ATP is required to meet the metabolic needs of
RBC and to fuel the ion pumps necessary for the
maintenance of the flexible, biconcave shape that
allows them to squeeze through narrow
capillaries.
• The anemia observed in glycolytic enzyme
deficiencies is a consequence of the reduced rate
of glycolysis, leading to decreased ATP
production by substrate-level phosphorylation.
Pyruvate reduction to lactate

• Lactate, formed from pyruvate by LDH,

is the final product of anaerobic
glycolysis in eukaryotic cells.
• Reduction to lactate is the major fate for
pyruvate in tissues that are poorly
vascularized (for example, the lens and
cornea of the eye and the kidney medulla)
or in RBC that lack mitochondria.
1. Lactate formation in muscle:
• In exercising skeletal muscle, NADH production (by glyceraldehyde 3-
phosphate dehydrogenase and by the three NAD+-linked dehydrogenases of
the TCA cycle) exceeds the oxidative capacity of the ETC.
• This results in an elevated NADH/NAD+ ratio, favoring reduction of pyruvate
to lactate by LDH.
• Therefore, during intense exercise, lactate accumulates in muscle, causing a
drop in the intracellular pH, potentially resulting in cramps.
• Much of this lactate eventually diffuses into the bloodstream and can be used
by the liver to make glucose

2. Lactate utilization:
• The direction of the LDH reaction depends on the relative intracellular
concentrations of pyruvate and lactate and on the ratio of NADH/NAD+.
For example, in the liver and heart, this ratio is lower than in exercising
muscle.
• Consequently, the liver and heart oxidize lactate (obtained from the blood) to
pyruvate.
• In the liver, pyruvate is either converted to glucose by gluconeogenesis or
converted to acetyl CoA that is oxidized in the TCA cycle.
• Heart muscle exclusively oxidizes lactate to carbon dioxide and water via the
TCA cycle.
3. Lactic acidosis:

• Elevated concentrations of lactate in the plasma, termed lactic acidosis (a type of metabolic
acidosis), occur when there is a collapse of the circulatory system, such as with myocardial
infarction, pulmonary embolism, and uncontrolled hemorrhage, or when an individual is in shock.
• The failure to bring adequate amounts of O2 to the tissues results in impaired oxidative
phosphorylation and decreased ATP synthesis.
Energy yield from glycolysis

1. Anaerobic glycolysis: A net of two molecules of ATP are generated for

each molecule of glucose converted to two molecules of lactate.
• There is no net production or consumption of NADH.

2. Aerobic glycolysis: The generation of ATP is the same as in anaerobic

glycolysis (that is, a net gain of two ATP per molecule of glucose).
• Two molecules of NADH are also produced per molecule of glucose.
• Ongoing aerobic glycolysis requires the oxidation of most of this NADH
by the ETC, producing three ATP for each NADH molecule entering the
chain
HORMONAL REGULATION

• Regular consumption of meals rich in carbohydrate or

administration of insulin initiates an increase in the amount of
glucokinase, PFK-1, and PK in the liver.
• Conversely, gene expression of the three enzymes is
decreased when plasma glucagon is high and insulin is low
(for example, as seen in fasting or diabetes).

Effect of insulin and glucagon on the expression of key

enzymes of glycolysis in the liver.
If insulin is insufficient, Fatty acids instead of Glucose activate TCA and thus blood
pH decreases.

• Normally, fasting blood glucose is

between 70-100 mg/dL (4.6-
6.1mmol/L).
• According to current information,
this value should not exceed ≥140
mg/dL (7.8 mmol/L) 2 hours after
a meal.
 Blood Glucose Rises in Insulin Deficiency (Diabetes Mellitus)

• In the case of insulin deficiency, symptoms such as weight loss,

polyuria, polydipsia are observed even if adequate nutrition is
provided.
• Insulin is the main hormone that prevents the rise of blood sugar in the
organism. It does this by regulating glucose-related metabolic
pathways.
• Insulin is also an anabolic hormone.
• It provides storage of carbohydrates and fats. It also shows its anabolic
effect by positively affecting protein synthesis.
• The tissues where insulin is mainly effective are muscle, liver and
adipose tissues.
ALTERNATE FATES OF PYRUVATE

• Pyruvate can be metabolized to products other than lactate.

1. Oxidative decarboxylation to acetyl CoA
• PDHC irreversibly converts pyruvate, the end product of aerobic
glycolysis, into acetyl CoA, a TCA cycle substrate and the carbon
source for fatty acid synthesis.
2. Carboxylation to oxaloacetate
• Carboxylation of pyruvate to oxaloacetate by pyruvate carboxylase
is a biotin-dependent reaction. This irreversible reaction is important
because it replenishes the TCA cycle intermediate and provides
substrate for gluconeogenesis.
3. Reduction to ethanol (microorganisms)
• The decarboxylation of pyruvate to acetaldehyde by thiamine-
requiring pyruvate decarboxylase occurs in yeas and certain other
microorganisms but not in humans.
Relationship between Glycolysis and Erythrocyte

• Erythrocytes are cells with a special structure that carry oxygen from the lungs to the tissues and
carry waste materials such as CO2 from the tissues to the lungs.
• A few special molecules are required for erythrocytes to release the oxygen they carry from the
lungs to the tissues or to do the opposite.
• 2,3diphosphoglycerate (2,3DPG) is one of them.
• The amount of this molecule is one of the important factors that enable the oxygen carried from the
air to be released to the tissues.
Glycolysis is required for the Bohr Curve showing Oxygen Saturation. Because
glycolysis is where 2,3 diphosphoglycerate is produced.

• Saturation of Hb with O2 is related to the O2

concentration of the environment (partial oxygen
pressure = pO2 ).
• The saturation curve of hemoglobin is sigmoidal.
(Allosteric modulators: 2,3-BPG, hydrogen ions)
BOHR EFFECT

• The Bohr effect is the oxygen binding curve of

hemoglobin under a certain pressure.
• In the formation of this curve;
• The amount of CO2
• pH
• Some glycolytic enzymes
• 2,3,DPG are important.
• The curve has a normal position.
• Formation above or below this causes difficulty
in transporting oxygen and releasing it to the
tissues.
• For these reasons, the curve showing the Bohr effect has physiological significance.
• When arterial blood enters the tissues, CO2 enters the erythrocytes by diffusion. Meanwhile, the pH
decreases. These conditions reduce the affinity of hemoglobin for oxygen.
• As a result, oxygen is released into the cell environment.
• H+HbO2 → O2+HHb
• Reactions in the lungs take place in contrast to those in the tissues.
• Here, the pH rises with the loss of CO2.
• In this case, the affinity of hemoglobin for oxygen increases and the binding of oxygen to
hemoglobin increases at low pressure.
Glycosylated Hemoglobins

• Hemoglobin is the substance inside red blood cells that carries oxygen to the cells.
• Glucose molecules in the blood normally become stuck to hemoglobin molecules - this means the
hemoglobin has become glycosylated (also referred to as hemoglobin A1c, or HbA1c).
• As a person's blood sugar becomes higher, more of the person's hemoglobin becomes glycosylated.
• The glucose remains attached to the hemoglobin for the life of the red blood cell, or about 3
months.
• A blood test can measure the amount of glycosylated hemoglobin in the blood.
• The glycosylated hemoglobin test shows what a person's average blood glucose level was for the 3
months before the test.
• This help determine how well a person's diabetes is being controlled over time.
Thank your for your attention