Chapter 9

CHEMOTROPIC
ENERGY:GLYCOLYSIS AND
FERMENTATION
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Glycolysis Generates ATP by
Catabolizing Glucose to Pyruvate
• Glycolysis (or the glycolytic pathway) is a ten-
step reaction sequence that converts one
glucose molecule into two molecules of pyruvate

• Pyruvate is a three-carbon compound

• Both ATP and NADH are produced

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Figure 9-6

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Figure 9-7

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Glycolysis is present in all organisms
• Glycolysis is common to both aerobic and
anaerobic organisms

• In most cells the enzymes for glycolysis are

found in the cytosol

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Glycolysis in Overview
• In the absence of oxygen glycolysis leads to
fermentation

• In the presence of oxygen glycolysis leads to

aerobic respiration

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Important features of the glycolytic
pathway are
• The initial input of ATP (Gly-1)

• The sugar splitting reaction in which glucose is split

into two three-carbon molecules

• The oxidative event that generates NADH (Gly-6)

• The two steps at which the reaction sequence is

coupled to ATP generation (Gly-7 and Gly-10)
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The glycolytic pathway can be divided
into three phases
• Phase I: the preparatory and cleavage steps

• Phase II: the oxidative sequence, which is the

first ATP-generating event

• Phase III: the second ATP-generating event

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Phase I: Preparation and Cleavage
• The net result of the first three reactions is to
convert glucose into a doubly phosphorylated
molecule (fructose-1,6-bisphosphate)

• The phosphates are transferred to glucose from

ATP

• ATP hydrolysis is also the driving force that

makes the phosphorylation exergonic and thus
irreversible

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The first reaction adds a phosphate to the
sixth carbon atom
• The bond formed is a phosphodiester bond, a
lower-energy bond than the phosphoanhydride
bonds in ATP

• The enzyme that catalyzes the reaction is

hexokinase, and is specific for phosphorylation of
other six-carbon sugars as well

• Liver cells also have glucokinase, which is specific

for just glucose
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The second phosphate is added to
carbon one
• The first carbon of glucose is not as easily
phosphorylated as the sixth

• The Gly-2 reaction first converts glucose-6-

phosphate to fructose-6-phosphate (isomerase),
allowing the Gly-3 reaction to add a phosphate to
carbon one

• This reaction is catalyzed by the enzyme

phosphofructokinase-1 (PFK-1)
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Summary of Gly-1 to Gly-5
• Glucose is split into two 3-C compounds by
aldolase
• The first phase of the glycolytic pathway can
be summarized as

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Phase 2: Oxidation and ATP Generation
• The net energy yield of phase one is negative

• Two molecules of ATP have been consumed

per molecule of glucose

• In phase 2, ATP production is linked to an

oxidative event, followed by the generation of
ATP in phase 3

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Gly-6 and Gly-7
• The oxidation of glyceraldehyde-3-phosphate
to 3-phosphoglycerate is highly exergonic,
and drives

– the reduction of NAD+ to NADH (Gly-6)

– the phosphorylation of ADP with inorganic

phosphate, Pi (Gly-7)

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Important features of Gly-6 and Gly-7
• NAD+ is an electron acceptor

• The oxidation is coupled to the formation of a

high-energy, doubly phosphorylated
intermediate, 1,3-bisphosphoglycerate

• ATP generation by transferring a phosphate

group to ADP from a phosphorylated substrate
such as 1,3-bisphosphoglycerate is called
substrate-level phosphorylation

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Summary of Gly-6 and Gly-7
• Gly-6 and Gly-7 can be summarized as

• Each reaction involving glyceraldehyde-3-

phosphate occurs twice per starting molecule of
glucose

• The two ATPs invested in the first phase are

recovered in the second phase, for no net ATP
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Phase 3: Pyruvate Formation and
ATP Generation
• The phosphoester bond of 3-phosphoglycerate is
converted to a phosphoenol bond

• The phosphate group is moved to the adjacent

carbon, forming 2-phosphoglycerate (Gly-8)

• Water is removed from the 2-phosphoglycerate by

the enzyme enolase (Gly-9) generating the high-
energy compound phosphoenolpyruvate (PEP)
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Phosphoenolpyruvate
• Hydrolysis of the phosphoenol bond of PEP is
one of the most exergonic known in biological
systems

• PEP hydrolysis drives ATP synthesis by

transferring a phosphate to ADP, catalyzed by
the enzyme pyruvate kinase (Gly-10)

• The transfer is irreversible in the direction of

pyruvate and ATP formation

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Summary of Gly-8 to Gly-1
• The third phase of glycolysis can be summarized
as

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Summary of Glycolysis
• The two molecules of ATP formed in the second
phosphorylation event (Gly-10) represent the net
yield of ATP for the glycolytic pathway

• .

• The pathway is highly exergonic in the direction

of pyruvate formation; DG in a cell is typically
–20 kcal/mol

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Conservation of Glycolysis
• The glycolytic pathway is one of the most
common and highly conserved metabolic
pathways known

• Virtually all cells have the ability to convert

glucose to pyruvate, extracting energy in the
process

• The next steps depend on the availability of

oxygen

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Alternative Substrates for Glycolysis
• Glucose is a major substrate for both
fermentations and respiration in a variety of
organisms and some tissues

• But for some organisms and tissues, glucose is

not significant at all

• There are a variety of alternatives to glucose,

which are often converted into an intermediate in
the glucose catabolism pathway

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Other Sugars and Glycerol Are Also
Catabolized by the Glycolytic Pathway
• Many sugars are available to cells, either
monosaccharides or disaccharides that can be
readily hydrolyzed into monosaccharides

• The monosaccharides are then converted into a

glycolytic intermediate

• Glucose and fructose enter most directly after

phosphorylation on carbon atom 6; mannose and
fructose require more steps
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Pentoses and glycerol can be channeled
into the glycolytic pathway too
• Phosphorylated pentoses can enter the glycolytic
pathway but must first be converted to hexose
phosphates

• Glycerol, a three-carbon molecule resulting from lipid

breakdown, can enter the cycle

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Figure 9-9

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Polysaccharides Are Cleaved to Form
Sugar Phosphates That Also Enter
the Glycolytic Pathway
• Glucose occurs primarily in the form of storage
polysaccharides, most often starch in plants and
glycogen in animals

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Gluconeogenesis
• The process of glucose synthesis is called
gluconeogenesis

• Glucose is synthesized from three- and four-

carbon precursors (noncarbohydrate)

• Pyruvate and lactate are the most common

starting materials
• Occurs in all organisms and in animals mainly
in liver and kidneys
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Gluconeogenesis and glycolysis
• Gluconeogenesis occurs by simple reversal of
glycolysis using the same enzyme in both
directions

• But not all the steps are simple reversals of

glycolysis: Gly-1, Gly-3, and Gly-10 are
accomplished by other means

• These are the most exergonic reactions of

glycolysis
• Thermodynamically difficult to reverse
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Figure 9-11

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Gluconeogenesis

• Biosynthetic anabolic pathways are seldom

just a reversal of the corresponding catabolic
pathways
• Glucose to pyruvate has G about
-20kcal/mole
• Reverse process would be about
+20kcal/mole and highly endergonic and
thermodynamically impossible

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Gluconeogenesis

• Gly1, Gly3 and Gly10 do not simply run in

reverse
– Gluconeogenic pathway has bypass
reactions
– Gly1 and Gly3 you have hydrolytic cleaving
that liberated inorganic phosphate instead
of trying to synthesize ATP.
• Exergonic reaction with G= -3.3 kcal/mole

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Gluconeogenesis

• Gly 10 is bypassed by a two-reaction

sequence
– Both reactions are driven by hydrolysis of a
phosphoanydride bond
– One from ATP and one from GTP
– Carbon dioxide is added to pyruvate in a
carboxylation reaction
– Next, carboxyl group is removed by
decarboxylation to form PEP
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Glycolytic vs Gluconeogenesis

• Glycolysis is catabolic producing net two

ATPs per glucose
• Gluconeogenesis in anabolic requiring the
equivalent of six ATPs per molecule of
glucose synthesized
• Gluconeogenesis proceed exergonically in
the direction of glucose synthesis

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What is happening right now to the
sugars you consume?
• Enzymes begin to break down carbohydrates
in the mouth (sucrase, maltase, lactase, etc)
• Glucose, galactose, and fructose are
absorbed by intestinal epithelial cells.- enter
bloodstream
– Galactosemia
Main sugar in blood is glucose. Regulated

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Cori Cycle

• Skeletal muscle is main source of blood

lactate
• It can function in either the presence or the
absence of oxygen
• Liver is primary site of gluconeogenesis
• Lactate is converted to glucose and released
into the blood-Cori cycle

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The Regulation of Glycolysis
and Gluconeogenesis
• Cells have enzymes for both glycolysis and
gluconeogenesis, so the processes must be
regulated
• Spatial regulation keeps the two processes
confined to separate places in the body
– Muscle cells – glycolysis Liver- gluconeogenesis

• There is also temporal regulation in which the

two processes take place at different times in
one cell
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Regulation of Glycolysis and
Gluconeogenesis
• Both are regulated to function at rates that
are responsive to cellular and organismal
needs for their products. ATP or glucose
• Regulated in reverse manner
• Intracellular conditions know to stimulate one
pathway usually have an inhibitory effect on
the other.

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Key Enzymes in the Glycolytic and
Gluconeogenic Pathways Are Subject
to Alllosteric Regulation
• Allosteric regulation involves the interconversion
of an enzyme between two forms, one catalytically
active and the other inactive

• The enzyme will be active or not depending on

whether an allosteric effector is bound to the
allosteric site

• The effector might be an activator or inhibitor

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Figure 9-12

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Key regulatory enzymes of glycolysis
and gluconeogenesis
• For glycolysis the enzymes are hexokinase,
phosphofructokinase-1 (PFK-1), and pyruvate
kinase

• For gluconeogenesis they are fructose-1,6-

bisphosphatase, and pyruvate carboxylase

• Each of the enzymes is unique to its pathway so

the pathways can be regulated independently
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Glycolysis and gluconeogenesis are
reciprocally regulated
• AMP and acetyl CoA, the two effectors to which
both pathways are sensitive, have opposite
effects
• AMP activates glycolysis and inhibits
gluconeogenesis
– Low ATP and high AMP cell is low on energy.
Activate glycolysis. High ATP inhibit glycolysis.

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• Acetyl CoA activate gluconeogenesis but
inhibits glycolysis
• High Acetyl CoA has enough energy inhibit
glycolysis

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Fructose-2,6-Bisphosphate Is an
Important Regulator of Glycolysis
and Gluconeogenesis
• Fructose-2,6-Bisphosphate (F2,6BP) is the most
important regulator of both glycolysis and
gluconeogenesis- ATP phosphorylation of C-2

• Synthesis of F2,6BP is catalyzed by

phosphofructokinase-2 (PFK-2), not PFK-1

• F2,6BP activates the glycolytic enzyme (PFK-1)

that phosphorylates fructose-6-phosphate and it
inhibits FBPase that catalyzes the reverse reaction
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Figure 9-13

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Effect of cAMP on F2,6BP
• cAMP affects the F2,6BP concentration in two
ways

• 1. It inactivates the PFK-2 kinase activity

• 2. It stimulates the F2,6BP phosphatase

activity

• These two effects tend to decrease the

concentration of F2,6BP in the cell

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Effect of cAMP on hormone regulation
• cAMP level in cells is controlled primarily by
the hormones glucagon and epinephrine
(adrenaline)

• These cause an increase in cAMP

concentration, stimulating gluconeogenesis
when more glucose is needed

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Novel Roles for Glycolytic Enzymes
• Glycolysis is connected to other cell processes
• Hexokinase (Gly-1) is a transcriptional repressor
in yeast cells under high glucose levels
• Mammals have four isoforms of hexokinase
- One is expressed in highly catabolically active
tumor cells
- Another binds to mitochondria and helps
coordinate glycolysis with mitochondrial functions
• Many other examples exist

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Additional functions of other glycolytic
enzymes
• Glyceraldehyde-3-phosphate dehydrogenase
(Gly-6) and enolase (Gly-9) have DNA-binding
abilities

• They can act as transcriptional regulators

• They connect the glycolytic pathway with

processes such as cell division and programmed
cell death
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Cancer connections
• Phosphoglucoisomerase (PGI; Gly-2) is
involved in cell motility and migration during
cancer cell metastasis

• Metastasis: the release of cells from malignant

tumors into the bloodstream; these can form
secondary tumors throughout the body

• PGI stimulates cell proliferation and migration

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