INTEGRATED BASIC SCIENCES

MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024

GLYCOLYSIS & OXIDATION OF PYRUVATE GLYCOLYSIS CAN FUNCTION UNDER ANAEROBIC
BIOMEDICAL IMPORTANCE CONDITIONS
• most tissues have at least some requirements for • fermentation in yeast was similar to breakdown of
glucose glycogen in muscle
o brain
▪ requirement is substantial • muscle contracts under anaerobic conditions:
o glycogen disappears & lactate appears
even in prolonged fasting:
▪ it can meet no > 20% of its energy • when O2 is admitted:
needs from ketone bodies o aerobic recovery takes place & lactate is no
longer produced
glycolysis
• main pathway of glucose metabolism • muscle contraction occurs under aerobic conditions:
• occurs in cytosol o pyruvate as end product of glycolysis

• can function either aerobically or anaerobically • pyruvate is oxidized further to CO2 & water
o depending on availability of O2 and ETC (‘.’ of
presence of mitochondria)

• RBC lack mitochondria ‘.’ completely reliant on

glucose as metabolic fuel
o metabolize it by anaerobic glycolysis.

ability of glycolysis to provide ATP in absence of

O2:
✓ allows skeletal muscle to perform at very high levels
of work output when O2 supply is insufficient
✓ allows tissues to survive anoxic episodes

• heart muscle, adapted for aerobic performance:

o relatively low glycolytic activity
o poor survival under conditions of ischemia

• diseases in w/c enzymes of glycolysis (eg, pyruvate

kinase) are deficient mainly seen:
o as hemolytic anemias
o if defect affects skeletal muscle (eg,
phosphofructokinase), as fatigue

fast-growing cancer cells

• glycolysis proceeds at high rate

• form large amounts of pyruvate, reduced to lactate

and exported
o produces acidic local environment in tumor

• lactate is used for gluconeogenesis in liver

• when O2 is in short supply:
o energy-expensive process
o responsible for much of hypermetabolism o mitochondrial reoxidation of NADH formed
seen in cancer cachexia during glycolysis is impaired
o NADH reoxidized by reducing pyruvate to
lactate
• Lactic acidosis results from:
o impaired activity of pyruvate ▪ = permit glycolysis to continue
dehydrogenase, in thiamin (B1) deficiency
• price of glycolysis under anaerobic conditions:
o limits amount of ATP formed per mole of
glucose oxidized ‘.’ more glucose must be
metabolized

• yeast & other microorganisms:

o pyruvate formed in anaerobic glycolysis is not
reduced to lactate, but decarboxylated &
reduced to ethanol

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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024

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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024
REACTIONS OF GLYCOLYSIS CONSTITUTE MAIN in glycolysis:
PATHWAY OF GLUCOSE UTILIZATION 2. converted to fructose-6-phosphate by
overall equation for glycolysis from glucose to lactate: phosphohexose isomerase
o involves an aldose–ketose isomerization

3. followed by another phosphorylation catalyzed by

phosphofructokinase (phosphofructokinase-1)
• all enzymes of glycolysis are cytosolic
forming fructose 1,6-bisphosphate
o irreversible under physiological conditions
• glucose enters glycolysis by phosphorylation to
glucose-6-
o enzyme both inducible and subject to
o catalyzed by hexokinase
allosteric regulation
▪ inhibited allosterically by its product G6P
▪ major role in regulating rate of glycolysis
o ATP as phosphate donor
4. fructose 1,6-bisphosphate cleaved by aldolase
o irreversible
(fructose 1,6-bisphosphate aldolase) into 2 triose
phosphates
tissues other than liver (& pancreatic β-islet cells):
✓ glyceraldehyde-3-phosphate
• availability of glucose for glycolysis (or glycogen
✓ dihydroxyacetone phosphate
synthesis in muscle & lipogenesis in adipose tissue)
controlled by transport into cell
are interconverted by enzyme
o regulated by insulin
phosphotriose isomerase
hexokinase
5. glycolysis continues w/ oxidation of
• high affinity (low Km) for glucose
glyceraldehyde- 3-phosphate to 1,3-
• in liver: saturated under normal conditions
bisphosphoglycerate
• ‘.’ acts at constant rate to provide glucose-6-
o by glyceraldehyde-3-phosphate
phosphate to meet liver’s needs
dehydrogenase
• liver cells contain an isoenzyme of hexokinase:
▪ NAD dependent
o glucokinase
▪ consists of 4 identical polypeptides
(monomers) forming a tetramer
glucokinase
• Km > normal intracellular concentration of glucose
▪ 4—SH groups present on each polypeptide
from cysteine residues w/in polypeptide
• removes glucose from hepatic portal blood
chain
following meal
• one found @ active site of enzyme
o ‘.’ regulate concentration of glucose available to
peripheral tissues

• provides more glucose-6-phosphate than required

for glycolysis
o used for glycogen synthesis & lipogenesis

• also found in pancreatic β-islet cells

o detects high concentrations of glucose in portal
blood

• as more glucose is phosphorylated by glucokinase,

there is increased glycolysis
o = increased formation of ATP
o = closure of an ATP-K+ channel, causing
membrane depolarization & opening of voltage-
gated Ca++ channel

• resultant influx of Ca++

o fusion of insulin secretory granules w/ cell
membrane
o release of insulin

• glucose-6-phosphate: important compound @ Mechanism of oxidation of glyceraldehyde-3-phosphate.

junction of metabolic pathways (Enz, glyceraldehyde-3-phosphate dehydrogenase.) enzyme
✓ glycolysis is inhibited by—SH poison iodoacetate, thus able to inhibit
✓ gluconeogenesis glycolysis. NADH produced on enzyme is not so firmly bound to
enzyme as is NAD+ ‘.’ NADH is easily displaced by another
✓ pentose phosphate pathway
molecule of NAD+.
✓ glycogenesis
✓ glycogenolysis

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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES
o substrate initially combines w/ —SH group
▪ = thiohemiacetal oxidized to thiol ester
▪ hydrogens removed oxidation are
transferred to NAD+
o thiol ester then undergoes phosphorolysis
▪ inorganic phosphate (Pi) added
• = 1,3-bisphosphoglycerate
• = free —SH group
6. Phosphate is transferred from 1,3-
bisphosphoglycerate onto ADP
o catalyzed by phosphoglycerate kinase
forms:
o ATP (substrate-level phosphorylation)
o 3-phosphoglycerate
• 2 mol triose phosphate formed per molecule of
glucose metabolized
o = 2× ATP formed in this reaction per molecule
of glucose
• toxicity of arsenic
o result of competition of arsenate w/ inorganic
phosphate (Pi)
▪ = 1-arseno-3-phosphoglycerate
• undergoes spontaneous hydrolysis to
3-phosphoglycerate w/o forming ATP
7. 3-Phosphoglycerate is isomerized to 2-
phosphoglycerate
o by phosphoglycerate mutase
o likely that 2,3-bisphosphoglycerate
(diphosphoglycerate, DPG) is an intermediate
8. subsequent step is catalyzed by enolase &
involves a dehydration
o = phosphoenolpyruvate
enolase
o inhibited by fluoride
▪ blood samples taken for measurement of
glucose: glycolysis is inhibited by taking
sample into tubes containing fluoride
o dependent on presence of either Mg2+ or
Mn2+
10. phosphate of PEP is transferred to ADP in
another substrate-level phosphorylation
o by pyruvate kinase
o form 2× ATP per molecule of glucose oxidized
essentially irreversible under physiological
conditions because of:
o large free-energy change involved
o immediate product of enzyme-catalyzed
reaction is enolpyruvate
▪ undergoes spontaneous isomerization
to pyruvate ‘.’ product of reaction is not
available to undergo reverse reaction
MARCH | 2024
• availability of O2 now determines w/c of 2 pathways
is followed
under anaerobic conditions:
• NADH cannot be reoxidized through ETC
• pyruvate is reduced to lactate catalyzed by lactate
dehydrogenase
o = permits oxidization of NADH
▪ ‘.’ another molecule of glucose to undergo
glycolysis
under aerobic conditions:
• pyruvate is transported into mitochondria
o undergoes oxidative decarboxylation to acetyl-
CoA then oxidation to CO2 in citric acid cycle
• reducing equivalents from NADH formed in
glycolysis are taken up into mitochondria for
oxidation via either:
✓ malate-aspartate shuttle
✓ glycerophosphate shuttle
TISSUES THAT FUNCTION UNDER HYPOXIC
CONDITIONS PRODUCE LACTATE
• skeletal muscle, particularly white fibers
o rate of work output ‘.’ hence need for ATP
formation exceed rate at w/c O2 can be
taken up and utilized
glycolysis in erythrocytes
• always terminates in lactate
• subsequent reactions of pyruvate oxidation are
mitochondrial w/c they lack
other tissues that normally derive energy from
glycolysis & produce lactate:
✓ brain ✓ GI tract
✓ renal medulla ✓ retina
✓ skin
• lactate production
o increased in septic shock
o in many cancers
• liver, kidneys, & heart normally take up lactate
and oxidize it, but produce it under hypoxic
conditions.
• lactate production is high [e.g. vigorous exercise,
septic shock, & cancer cachexia]
o much is used in liver for gluconeogenesis
▪ = increase in metabolic rate to provide
ATP and GTP needed
o increase in O2 consumption from increased
oxidation of metabolic fuels to provide ATP and
GTP needed for gluconeogenesis
▪ = oxygen debt
• lactate may be formed in cytosol
o then enter mitochondrion to be oxidized to
pyruvate for onward metabolism
o provides a pathway for transfer of reducing
equivalents from cytosol into mitochondrion
for ETC in addition to glycerophosphate &
malate-aspartate shuttles
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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024
GLYCOLYSIS IS REGULATED AT 3 STEPS RBCS: 1 SITE OF ATP FORMATION IN
ST

INVOLVING NON-EQUILIBRIUM REACTIONS GLYCOLYSIS MAY BE BYPASSED

• most reactions of glycolysis are freely reversible • reaction catalyzed by phosphoglycerate kinase
w/ 3 markedly exergonic ‘.’ considered to be o may be bypassed by reaction of
physiologically irreversible bisphosphoglycerate mutase
▪ = catalyzes conversion of 1,3-BPG to 2,3-
rxns catalyzed by: BPG
✓ hexokinase (and glucokinase)
✓ phosphofructokinase • followed by hydrolysis to 3-phosphoglycerate and Pi,
✓ pyruvate kinase by 2,3-bisphosphoglycerate phosphatase

major sites of regulation of glycolysis

phosphofructokinase
• significantly inhibited @ normal intracellular
concentrations of ATP
o rapidly relieved by 5′AMP formed as ADP
begins to accumulate
▪ signal need for increased rate of glycolysis

• cells capable of gluconeogenesis (reversing

glycolytic pathway) have different enzymes that
catalyze reactions:
o to reverse irreversible steps:
▪ glucose-6-phosphatase
▪ fructose 1,6-bisphosphatase

o to reverse reaction of:

▪ pyruvate kinase
▪ pyruvate carboxylase
▪ phosphoenolpyruvate carboxykinase

• reciprocal regulation of phosphofructokinase in

glycolysis & fructose 1,6-bisphosphatase
o in gluconeogenesis

• fructose enters glycolysis by phosphorylation to

fructose-1-phosphate
o bypasses main regulatory steps
▪ = more pyruvate and acetyl-CoA than
required for ATP formation
• pathway involves NO NET yield of ATP from
glycolysis
in liver & adipose tissue:
o rather provides 2,3-BPG
o this leads to increased lipogenesis
▪ binds to hemoglobin
o ‘.’ high intake of fructose may be a factor in
▪ decreasing its affinity for oxygen ‘.’
development of obesity
oxygen more readily available to tissues

OXIDATION OF PYRUVATE TO ACETYL-COA IS THE

IRREVERSIBLE ROUTE FROM GLYCOLYSIS TO
CITRIC ACID CYCLE
• pyruvate into mitochondrion: proton symporter

• then undergoes oxidative decarboxylation to

acetyl-CoA by a multienzyme complex associated
w/ inner mitochondrial membrane
o pyruvate dehydrogenase complex
o analogous to α-ketoglutarate dehydrogenase
complex of citric acid cycle

• pyruvate is decarboxylated by pyruvate

dehydrogenase to a hydroxyethyl derivative of
thiazole ring of enzyme-bound thiamin diphos-
phate
o in turn reacts w/ oxidized lipoamide [prosthetic
group of dihydrolipoyl transacetylase]
▪ = acetyl lipoamide

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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024
PYRUVATE DEHYDROGENASE IS REGULATED BY
END-PRODUCT INHIBITION & COVALENT
MODIFICATION
• pyruvate dehydrogenase is inhibited by its
products
o = acetyl-CoA & NADH

Oxidative decarboxylation of pyruvate by the pyruvate

dehydrogenase complex. Lipoic acid joined by an amide link to
lysine residue of transacetylase component of enzyme complex. It
forms a long flexible arm, allowing lipoic acid prosthetic group to
rotate sequentially between active sites of each enzyme of
complex. (FAD, flavin adenine dinucleotide; NAD+, nicotinamide
adenine dinucleotide.)

thiamin (vitamin B1) deficiency

• glucose metabolism is impaired Regulation of pyruvate dehydrogenase (PDH). Arrows with
• significant (& potentially life-threatening) lactic and wavy shafts indicate allosteric effects. (A) Regulation by end-
product inhibition. (B) Regulation by interconversion of active and
pyruvic acidosis inactive forms.

• acetyl lipoamide reacts w/ coenzyme A • regulated by:

o = acetyl-CoA & reduced lipoamide o phosphorylation (by kinase) of 3 serine
residues on pyruvate dehydrogenase
• reaction is completed when reduced lipoamide is component of multienzyme complex
reoxidized by flavoprotein, dihydrolipoyl ▪ decreased activity
dehydrogenase
o contain flavin adenine dinucleotide (FAD) o dephosphorylation (by phosphatase)
▪ increase in activity
• reduced flavoprotein is oxidized by NAD+
o in turn transfers reducing equivalents to • kinase is activated by increases in ratios of:
respiratory chain ✓ [ATP]/[ADP]
✓ [acetyl-CoA]/[CoA]
overall rxn: ✓ [NADH]/[NAD+]

• ‘.’ pyruvate dehydrogenase, and ‘.’ glycolysis, is

both inhibited by:
• complex consists polypeptide chains of each 3
✓ adequate ATP (& reduced coenzymes for ATP
component enzymes
formation) available
o intermediates do not dissociate, but channeled
✓ fatty acids being oxidized
from one enzyme site to next
o ‘.’ increases rate of reaction & prevents side
in fasting:
reactions
• when nonesterified fatty acid concentrations
increase = decrease of enzyme in active form
o = spare carbohydrate

in adipose tissue:
• glucose provides acetyl-CoA for lipogenesis
• ‘.’ enzyme is activated in response to insulin

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INTEGRATED BASIC SCIENCES
MODULE 11 | METABOLISM OF CARBOHYDRATES MARCH | 2024
CLINICAL ASPECTS
INHIBITION OF PYRUVATE METABOLISM LEADS
TO LACTIC ACIDOSIS
• arsenite & mercuric ions
o react w/ —SH groups of lipoic acid
o inhibit pyruvate dehydrogenase, as does a
dietary deficiency of thiamin
o = pyruvate accumulation

• many alcoholics are thiamin deficient (poor diet +

alcohol inhibiting thiamin absorption)
o develop potentially fatal pyruvic & lactic
acidosis

• patients w/ inherited pyruvate dehydrogenase

deficiency
o result of defects in components of enzyme
complex

o present w/ lactic acidosis

▪ after a glucose load

because of brain’s dependence on glucose fuel:

o metabolic defects commonly cause neurological
disturbances

• inherited aldolase A deficiency & pyruvate

kinase deficiency in RBCs
o cause hemolytic anemia

• exercise capacity of patients w/ muscle

phosphofructokinase deficiency is low
o particularly if on high-carbohydrate diets