Chapter VIII –

Carbohydrates
Metabolism
WALLY I. TAPAS, RPh, MA (Cand.)
 Learning
Outcomes
LEARNING OUTCOMES:
At the end of this lecture, students are expected to:
1. Recall the concepts of Carbohydrate metabolism and integrate
Metabolism is the sum of each specific pathways.
all the chemical reactions 2. Correlate the fates of pyruvate to establish interconnection
in a cell, tissue, or the between metabolic pathways.
whole body. 3. Understand the pathophysiology of clinical diseases that root
from faulty carbohydrate metabolism.
ANABOLIC
Synthesis of compounds from smaller raw materials
• Endergonic and divergent process (+ΔG)
• Examples: Protein and Triglyceride Synthesis, Glycogenesis

CATABOLIC
• Breakdown of larger molecules
• Usually oxidative reactions
• Exergonic and convergent process
• Produces reducing equivalent & ATP mainly via respiratory chain
• Examples: Glycolysis, Beta-Oxidation, Glycogenolysis

AMPHIBOLIC
• Crossroads of metabolism
• Link between anabolic and catabolic pathways
• Example: Citric Acid Cycle
REGULATORS OF METABOLISM
1. INTRACELLULAR
• Signals from within the cell
• Substrate availability and product inhibition
• Allosteric activators/inhibitors

2. INTERCELLULAR
• Communication between cells
o Direct contact (Gap junctions)
o Synaptic signaling (Neurotransmitters)
o Endocrine signaling (Hormones)
 GLYCOLYSIS

Major pathway for glucose metabolism

Converts glucose into 3 carbon compounds to
provide energy
Most common: Embden-Mayerhof-Parnas Pathway
• Occurs in: cytosol of all mammalian cells
• Substrate: Glucose
• End-products: 2 pyruvate or lactate
• Rate-limiting step:
Fructose-6-phosphate→Fructose-1,6-bisphosphate
Enzyme: Phosphofructokinase-1
IMPORTANT STEPS OF GLYCOLYSIS
STEP 1:PHOSPHORYLATION OF GLUCOSE
Glucose → Glucose-6-phosphate
Enzyme: Hexokinase/Glucokinase

STEP 3: PHOSPHORYLATION OF FRUCTOSE-6-PHOSPHATE

Enzyme: Phosphofructokinase-1 (PFK-1)

STEP 10: FORMATION OF PYRUVATE

Phosphoenolpyruvate → Pyruvate + ATP
• Enzyme: Pyruvate kinase
• Substrate-level phosphorylation
o Yields 1 ATP per molecule of PEP = 2
• Regulation
o Activated by fructose-1,6-BP
o Inhibited by glucagon (Fructose 1,6-BP)
ATP PRODUCTION
2 steps in glycolysis produce ATP via substrate-level
phosphorylation
1,3-Bisphosphoglycerate → 3-Phosphoglycerate
Enzyme: Phosphoglycerate kinase
Phosphoenolpyruvate → Pyruvate
Enzyme: Pyruvate kinase

NADH PRODUCTION
• 1 step in glycolysis produces NADH
• This is an oxidation reaction that passes electrons to NAD+
to make NADH
Glyceraldehyde-3-Phosphate → 1,3 Bisphosphoglycerate
Enzyme: Glyceraldehyde-3-phosphate dehydrogenase
Aerobic vs Anaerobic
AEROBIC GLYCOLYSIS
• NADH proceeds to the electron transport chain (Complex I)
• NADH cannot pass through the inner mitochondrial membrane,
requires shuttles for transport.

ANAEROBIC GLYCOLYSIS
Enzyme: Lactate dehydrogenase
• NADH is used to reduce pyruvate to lactate
• Major fate of pyruvate in lens and cornea of the eye, kidney medulla,
testes, mature RBCs and WBCs
• MI and stroke
• Lactic acidosis happens in vigorous
exercise, septic shock, and
cancer cachexia
Fates of Pyruvate
2,3-BISPHOSPHOGLYCERATE
• Found in RBCs where the reaction catalyzed by
phosphoglycerate kinase is bypassed
• Rapoport-Luebering shunt pathway
1,3-BPG → 2,3-BPG
Enzyme: Bisphosphoglycerate mutase
ACETYL CoA PRODUCTION

PYRUVATE to ACETYL CoA

• Coenzymes:
o Thiamine pyrophosphate (from vitamin B1)
o FAD (from vitamin B2)
o NAD+ (from vitamin B3)
o Coenzyme A (contains pantothenic acid) (from vitamin B5)
o Lipoic acid (antioxidant)
MATURITY ONSET DIABETES OF THE YOUNG TYPE2
• Due to mutations that decrease the activity of
glucokinase
• Patients are generally asymptomatic, and
hyperglycemia is commonly discovered during routine
screening
• Majority of these patients do not require treatment,
except during pregnancy

ARSENIC POISONING
• Pentavalent arsenic competes with inorganic phosphate
as a substrate for glyceraldehyde-3-P dehydrogenase,
inhibits substrate level phosphorylation
o Final common pathway for the aerobic oxidation of
carbohydrates, lipids, and proteins
o Major pathway for formation of ATP
o Also provides substrates for gluconeogenesis,
amino acid synthesis, and fatty acid synthesis
o Occurs in: Mitochondrial matrix except: succinate
dehydrogenase (inner mitochondrial membrane)
• Substrate: Acetyl CoA
• Products: 2 CO2, 1 GTP, 3 NADH, and 1 FADH2
• Rate-limiting step: Isocitrate → α-ketoglutarate
Enzyme: Isocitrate dehydrogenase
TCA INTERMEDIATES
• Citrate: Delivers acetyl CoA to the cytosol for fatty
acid synthesis via the citrate shuttle
• Succinyl CoA: Used for heme synthesis and
activation of ketone bodies in extrahepatic tissues
• Malate: May be used for gluconeogenesis

REMEMBER THE FOLLOWING:

• The cycle does not synthesize new oxaloacetate
• There is no hormonal control
• Four B vitamins are essential in the TCA
• Anaplerotic reactions replenish intermediates of
the TCA that have been used for biosynthesis of
glucose, fatty acids, amino acids, or other
compounds
o Most important is pyruvate carboxylase reaction,
which maintains adequate concentration of OAA
o Other anaplerotic substrates: glutamine, glutamate,
aspartate, propionyl CoA
 GLUCONEOGENESIS
The process of synthesizing glucose from non-carbohydrate precursors,
in order to prevent hypoglycemia during a fast
Occurs in:
o Liver (90%) and the kidney (10%)
o In both mitochondria and cytosol (HUG)
Substrates:
1. Intermediates of glycolysis and TCA (except Acetyl CoA)
2. Lactate through the Cori Cycle
3. Glycerol and propionyl CoA from triacylglycerols
4. Carbon skeletons of glucogenic amino acids (CHONs)
Rate limiting step:
Fructose-1,6,-bisphosphate → Fructose-6-phospate
Enzyme: Fructose-1,6-bisphosphatase
GLUCONEOGENESIS: Important Steps
STEPS 1 & 2: PYRUVATE → OAA → PEP
• Pyruvate → Oxaloacetate
• Enzyme: Pyruvate carboxylase
o Requires biotin and ATP
o Allosterically activated by acetyl CoA
o Oxaloacetate is reduced to malate and exported from the mitochondrion to the
cytosol, where it is converted back to oxaloacetate
Oxaloacetate → Phosphoenolpyruvate
• Enzyme: Phosphoenolpyruvate Carboxykinase
o Requires GTP
o Key enzyme that catalyzes net transfer out of the citric acid cycle into gluconeogenesis
STEP 9: FRUCTOSE-1,6-BP → FRUCTOSE-6-P
• Enzyme: Fructose-1,6-bisphosphatase
o Rate-limiting step of gluconeogenesis
o Inhibited by fructose-2,6-bisphosphate and AMP
STEP 11: GLUCOSE-6-PHOSPHATE → GLUCOSE
• Enzyme: Glucose 6-phosphatase
o Final step shared with glycogenolysis
o Present in liver and kidney, to release glucose into the bloodstream
REGULATION OF GLUCONEOGENESIS
Gluconeogenesis is regulated primarily by:
o Circulating levels of glucagon
o Availability of glucogenic substrates
o Allosteric activation of hepatic pyruvate carboxylase by acetyl CoA
o Allosteric inhibition of fructose-1,6-bisphosphatase by AMP

ENERGY REQUIREMENT
• Gluconeogenesis from pyruvate requires:
o Cleavage of 6 high-energy phosphate bonds (4 ATP + 2 GTP)
o Oxidation of 2 NADH
• Energy is derived from the oxidation of fatty acids
Clinical Correlates
GLUCOSURIA
• In hyperglycemia, the glomerular filtrate may contain more glucose than can be reabsorbed
• Occurs when the venous blood glucose concentration exceeds 10.0 mmol/L (renal threshold)

HYPERGLYCEMIA IN CRITICALLY ILL PATIENTS

• Excessive gluconeogenesis in response to injury and infection
• Associated with poor outcome

HYPOGLYCEMIA DURING PREGNANCY

• High fetal glucose consumption
• Risk of maternal and fetal hypoglycemia especially during fasting

HYPOGLYCEMIA IN NEONATES
• Enzymes of gluconeogenesis are not yet fully developed
• Premature and low-birth-weight infants more susceptible because
they have little adipose tissue
• Major storage carbohydrate in animals
• Approximately 500 g of glycogen stored in muscle
and liver
• Branched polymer of α-D-glucose
o Primary glycosidic bond: α(1→4)
o Branch points after 8 to 10 residues: α(1→6)

GLUCOGENESIS
• Synthesis of glycogen
• Occur in: Liver and muscle Cytosol
• Substrate: α-D-glucose
• Rate-limiting Step: Elongation of glycogen
chains i.e., creation of α-(1,4) glycosidic bonds
Enzyme: Glycogen synthase
 GLYCOGENIN
• Protein that serves as a primer for glycogen synthesis when glycogen is completely depleted

• Otherwise, residual glycogen fragment can accept

glucose residues

SYNTHESIS OF UDP-GLUCOSE
• α-D-glucose is attached to uridine diphosphate and
this becomes the source of all glucosyl residues that are
added to the glycogen molecule
• Enzymes: phosphoglucomutase and UDP-glucose
pyrophosphorylase

ELONGATION OF GLYCOGEN CHAINS

• Enzyme: Glycogen synthase
• Forms α(1→4) bonds between glucose residues
• Bonds formed at the non-reducing end (i.e., carbon 4)
• The rate-limiting step of glycogenesis
ELONGATION OF GLYCOGEN CHAINS
• Enzyme: Glycogen synthase
• Forms α(1→4) bonds between glucose residues
• Bonds formed at the non-reducing end (i.e., carbon 4)
• The rate-limiting step of glycogenesis

FORMATION OF BRANCHES IN GLYCOGEN

• Enzyme: Branching enzyme composed of amylo α(1→4) →α(1→6) transglucosidase
• Forms new α(1→6) bonds by transferring 5 to 8 glucosyl residues
GLYCOGENOLYSIS
• Mobilizing stored glycogen
• Occurs in: Liver and muscle, Cytosol
• Substrate: Glycogen
• Products:
o Glucose in liver
o Glucose-6-phosphate in muscle
• Rate-limiting Step: Shortening of glycogen chains by Glycogen phosphorylase

SHORTENING OF CHAINS
• Sequential cleavage of α(1→4) bonds between the glucosyl
residues at the non-reducing ends of the glycogen chains
• Enzyme: Glycogen phosphorylase
• Coenzyme: Pyridoxal phosphate
• Stops when only 4 glucosyl units remain (limit dextrin)
REMOVAL OF BRANCHES
• Enzyme: Debranching enzyme composed of
o α(1→4) → α(1→4) glucantransferase
o Amylo-α(1→6) glucosidase
• Involves cleavage of α(1→4) and α(1→6) bonds
• Yields free glucose from cleavage of the α(1→6) bond

GLUCOSE-1-P → GLUCOSE-6-P → GLUCOSE

• Enzymes: phosphoglucomutase and, when present, glucose-6- phosphatase
• End product in muscle: glucose-6-phosphate
• End product in liver: glucose

LYSOSOMAL DEGRADATION OF GLYCOGEN

• Enzyme: α(1→4) glucosidase (acid maltase)
• Degrades 1 to 3% of glycogen
• Purpose unknown, but deficiency of enzyme activity leads to Pompe disease
 OTHER
CARBOHYDRATE
METABOLISM
PHOSPHORYLATION OF FRUCTOSE
• Fructose → Fructose-1-P
• Enzyme: Fructokinase or hexokinase
FORMATION OF DHAP AND
GLYCERALDEHYDE
• Fructose-1-P → DHAP + Glyceraldehyde
• Enzyme: Aldolase B
Aldolase A – most tissues
PHOSPHORYLATION OF GALACTOSE
• Galactose → Galactose-1-P
• Enzyme: Galactokinase
FORMATION OF UDP-GALACTOSE
• Galactose-1P + UDP-glucose→ UDP-galactose+Glucose-1P
Enzyme: Galactose-1-P uridyl transferase (GALT)
USE OF GALACTOSE AS CARBON SOURCE
• UDP-galactose → UDP-glucose
• Enzyme: UDP-hexose-4-epimerase
Conversion of glucose to sorbitol
• Glucose → Sorbitol
• Enzyme: Aldose reductase
o Found in the lens, retina, Schwann cells,
liver, kidney, placenta, red blood cells,
ovaries, and seminal vesicles

Conversion of sorbitol to fructose

• Sorbitol → Fructose
• Enzyme: Sorbitol dehydrogenase
o Found in the liver, ovaries and seminal
vesicles
 URONIC ACID PATHWAY
• Alternative pathway for oxidation of glucose in the liver
• Does not produce ATP
• Main pathway for production of glucuronic and iduronic acid
• Also synthesizes ascorbic acid in plants and mammals
o Not possible for guinea pigs and primates because of absence of
L-gulonolactone oxidase

GLUCURONIC ACID
• Essential component of
glycosaminoglycans (GAG)
• Also required in detoxification
reactions of insoluble compounds
o Bilirubin, steroids, morphine and
other drugs
• AKA Hexose MonoPhosphate shunt (HMP)
o Produces NADPH
o Produces ribose 5-phosphate required for biosynthesis of nucleotides
o Provides a mechanism for the metabolic use of 5-carbon sugars
o Take Note: Neither produces nor consumes ATP
• Occurs in:
o RBCs and tissues that produce lipids (liver, adipose tissue,
adrenals, thyroid, testes, lactating mammaries)
o In the cytosol
• Substrate: Glucose-6-phosphate
• Important products: NADPH and Ribose-5-phosphate
Rate-limiting step: Glucose-6-phosphate → 6-phosphogluconate
Enzyme: Glucose-6-phosphate dehydrogenase
 NADPH
o Reductive biosynthesis of fatty acids and steroids
o Glutathione reduction inside RBCs
o Cytochrome P450 monooxygenase system
o Oxygen-dependent bactericidal mechanism of WBCs
o Synthesis of nitric oxide

GLUTATHIONE
• Reduced glutathione (G-SH) removes H2O2 in a reaction catalyzed by
glutathione peroxidase
• Reduced glutathione is regenerated by glutathione reductase, which
requires NADPH
GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PD) DEFICIENCY
• Involves ↓NADPH in RBCs and ↓activity of glutathione reductase causing free radicals
and peroxides accumulate
• Presents with hemolytic anemia after oxidative stress, due to poor RBC defense against
oxidizing agents
• Precipitating factors:
o Infection (most common), drugs (sulfonamides, primaquine, chloramphenicol), Fava beans
• Pathology
o Heinz bodies - Altered hemoglobin that precipitates within RBCs
o Bite cells - Abnormally shaped RBCs due to phagocytic removal of Heinz bodies in spleen
CHRONIC GRANULOMATOUS DISEASE
• Deficiency in NADPH oxidase
o Converts molecular oxygen into superoxide in leukocytes
(especially neutrophils and macrophages) and used in the
respiratory burst
that kills bacteria
• Severe, persistent, and chronic pyogenic infections
caused by catalase-positive bacteria
This STRESSFUL presentation is prepared by:
Wally “the stress but fresh” Tapas