Glycolysis and Metabolic Fates of Pyruvate: Prof. Dr. Gerhard Grüber

Glycolysis and Metabolic fates of pyruvate
Prof. Dr. Gerhard Grüber

Nanyang Technological University
School of Biological Sciences
Division of Structural & Computational Biology
Singapore 637551
GGrueber@ntu.edu.sg
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State of reduction of carbon atoms in biomolecules
More reduced state Less reduced state
Fats Carbohydrates Carbonyls Carboxyls Carbon

dioxide
In living systems most of the energy needed to drive biosynthetic reactions is derived from the oxidation of
organic substrates. Oxygen, the ultimate electron acceptor for aerobic organisms, is a strong oxidant; it has the
tendency to attract electrons, becoming reduced in the process, e.g.:
becomes oxidized
becomes reduced
Garett & Grisham: Biochemistry 4th edition

The energy levels of electrons
Energy is the capacity to cause change
Potential energy is the energy that matter has because of its location or structure
The electrons of an atom have potential energy due to their distance from the nucleus
Changes in potential energy occur in steps of fixed amounts
An electron s energy level is correlated with its average distance from the nucleus
A ball bouncing down a flight

of stairs can come to rest only
on each step, not between steps.
Third shell (highest energy

level in this model)
Second shell (higher Energy

energy level) absorbed
First shell (lowest

Energy level)
Energy
lost
Atomic
nucleus
© 2017 Pearson Education, Ltd.

Stepwise Energy Harvest via NAD+
becomes oxidized
becomes reduced
In cellular respiration, glucose and other organic molecules are broken down in a series of steps.
Electrons from organic compounds are usually first transferred to NAD+, a coenzyme
2e 2H
2e H
NAD NADH H
Dehydrogenase
Reduction of NAD
2[H] H
(from food) Oxidation of NADH
Nicotinamide Nicotinamide
(oxidized form) (reduced form)

Overview of carbohydrate digestion
The major carbohydrates in the diet are starch,

lactose, and sucrose. The starches amylose and
amylopectin are polysaccharides composed of
hundreds to millions of glucosyl units linked
together through -1,4- and -1,6-glycosidic
bonds (Fig. right). Lactose is a disaccharide
composed of glucose and galactose, linked
together through a -1,4-glycosidic bond.
Sucrose is a disaccharide composed of glucose
and fructose, linked through an -1,2-glycosidic
bond. The digestive processes convert all of
these dietary carbohydrates to their constituent
monosaccharides by hydrolyzing glycosidic
bonds between the sugars.
Glucose is the universal fuel for human cells
and the source of carbon for the synthesis of
most other compounds. Every human cell type
uses glucose to obtain energy. The release of
the hormones insulin and glucagon by the
pancreas aids in the use and storage of 4)
✗ CI
Glucose
-
glucose. Other dietary sugars (mainly fructose +

and galactose) are converted to glucose or to ✗ (1-6) Galactose
intermediates of glucose metabolism. These pcl
-
4)
monosaccharides are absorbed from the
intestine, enter the blood, and travel to the sucrose + fructose
tissues, where they are metabolized (Fig. right).
↳ Glucose✗ ( 112)
C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition
Overview of carbohydrate metabolism
he ✗ 0 kinase
After glucose is transported into cells, it is

phosphorylated by a hexokinase to form glucose 6-
phosphate. Glucose 6-phosphate can then enter
several metabolic pathways. The three that are
common to all cell types are glycolysis, the pentose
phosphate pathway, and glycogen synthesis (Fig.
right).
Important. The phosphate group in glucose 6-
phosphate (G6-P) is completely ionized at
physiological pH, giving it an overall negative charge.
Major pathways of glucose metabolism.
Since the plasma membrane is impermeable to
charged molecules, G6-P cannot enter into the cells
from the blood stream. Hence phosphorylation of glucose prevents it from
leaving cells .
In tissues, fructose and galactose are

converted to intermediates of glucose
metabolism. Thus, the fate of these sugars
parallels that of glucose (Fig. right).
Overview of fructose and galactose metabolism.

Overview of glycolysis and the TCA cycle
I
10 reactions
can occur w/o oxygen

>
Glycolysis (from the Greek glyk-, sweet, and lysis, splitting) begins
with the phosphorylation of glucose to glucose 6 phosphate (glucose
6-P) by hexokinase. In subsequent steps of the pathway, one glucose
6-P molecule is oxidized to two pyruvate molecules with generation
of two molecules of nicotinamide adenine dinucleotide (NADH).
A net generation of two molecules of ATP occurs through direct 2X
transfer of high-energy phosphate from intermediates of the pathway
to adenosine diphosphate (ADP) (substrate-level phosphorylation).
Glycolysis occurs in the cytosol and generates cytosolic NADH.
Because NADH cannot cross the inner mitochondrial membrane, its
reducing equivalents are transferred to the electron-transport chain by
either the malate aspartate shuttle or the glycerol 3-phosphate
shuttle.
Pyruvate is then oxidized completely to CO2 by pyruvate
dehydrogenase and the tricarboxylic acid (TCA) cycle inside the
mitochondrion. Complete aerobic oxidation of glucose to CO2 can
generate approximately 30 to 32 mol of ATP per mole of glucose.

Energy Investment Phase
R
Glyceraldehyde
3-phosphate (G3P)
ATP Glucose Fructose ATP Fructose

Glucose 6-phosphate 6-phosphate 1,6-bisphosphate
ADP ADP
Isomerase
Hexokinase Phosphogluco- Phospho- Aldolase

Dihydroxyacetone
isomerase fructokinase
phosphate (DHAP)
Glucose metabolism begins with transfer of a phosphate from ATP to glucose to form
glucose 6-P. Phosphorylation of glucose commits it to metabolism within the cell because
glucose 6-P cannot be transported back across the plasma membrane. The phosphorylation
reaction is irreversible under physiologic conditions because the reaction has a high-
negative free, 0 . Phosphorylation does not, however, commit glucose to glycolysis.
This reaction of the first ATP investment involves nucleophilic attack of the C6-OH of
glucose on the electrophilic terminal ( ) phosphate of ATP. Magnesium ion is required
because the reactive form of ATP is its chelated complex with Mg2+. Mg2+ partially
neutralizes the negative charges of the oxygen atoms, making the -phosphorus atom more
accessible for nucleophilic attack, and thus a better electrophile.
Never forget me!

D. R. Appling, S. Anthony-Cahill, C. K. Mathews, Biochemistry: Concepts and Connections, 2nd edition (2019)
Energy Investment Phase
Glyceraldehyde
3-phosphate (G3P)

ADP ADP
Isomerase

Dihydroxyacetone
phosphate (DHAP)
Hexokinases, the enzymes that catalyze the phosphorylation of glucose, are a family of
tissue-specific isoenzymes that differ in their kinetic properties. The iso-enzyme found in
liver and -cells of the pancreas has a much higher Km than other hexokinases and is called
glucokinase. In many cells, some of the hexokinase is bound to porins in the outer
mitochondrial membrane, which gives these enzymes first access to newly synthesized ATP
as it exits the mitochondria.
Glucose 6-P is isomerized to fructose 6-phosphate (fructose 6-P) and subsequently cleaved
into two three-carbon fragments. The isomerization is essential for the subsequent cleavage
of the bond between carbons 3 and 4.
well enzyme binds to substrate
Km value how
.
"
lower the km value better the
binding
speed
, .
conch still binds well nigh catalyst ng

Hexo kinase → low km value
i. e low glucose ,
9]
Hero kinase
everything compared
to
km value [
Glucokinase →
higher

Isomerization of glucose 6-phosphate to fructose 6-phosphate
1
Never forget me!
2 2
Recall that the open-chain form of glucose has an aldehyde group at carbon 1, whereas the open-chain form
of fructose has a keto group at carbon 2. Thus, the isomerization of glucose 6-phosphate to fructose 6-
phosphate is a conversion of an aldose into a ketose. The reaction catalyzed by phosphoglucose isomerase
includes additional steps because both glucose 6-phosphate and fructose 6-phosphate are present primarily in
the cyclic forms. The enzyme must first open the six-membered ring of glucose 6-phosphate, catalyze the
isomerization, and then promote the formation of the five-membered ring of fructose 6-phosphate.
In summery, the isomerization reaction is necessary because
the of the carbonyl from C-1 to C-2 creates a new primary alcohol function
at C-1, which becomes easily phosphorylated,
activation of C-3, facilitating the C-C bond cleavage in the 4th step of glycolysis.
J. M. Berg, J. L. Tymoczko & L. Stryer, Biochemistry 7th edition

Reaction of the PFK-1 within the Energy Investment Phase
Glyceraldehyde
3-phosphate (G3P)

ADP ADP
Isomerase

Dihydroxyacetone
phosphate (DHAP)
The next step of glycolysis (reaction 3) is the phosphorylation of fructose 6-P to fructose 1,6-bisphosphate (fructose
1,6-bisP). The reaction involves the same nucleophilic substitution chemistry we saw for the hexokinase (reaction
1). Here in reaction 3, the C1-OH of F6P is the nucleophile that attacks the electrophilic -phosphate of ATP. The
reaction catalyzed by phosphofructokinase-1 (PFK-1), is generally considered the first committed step of the
pathway and is thermodynamically and kinetically irreversible. Therefore, PFK-1 irrevocably commits glucose to
the glycolytic pathway. PFK-1 is a regulated enzyme in cells, and its regulation controls the entry of glucose into
glycolysis. Like the hexokinase, it exists as tissue-specific isoenzymes whose regulatory properties match variations
in the role of glycolysis in different tissues.

Aldol cleavage within the Energy Investment Phase
Glyceraldehyde
3-phosphate (G3P)

ADP ADP
Isomerase

Dihydroxyacetone
phosphate (DHAP)
Fructose 1,6-bisP is cleaved into two phosphorylated three-carbon

compounds (triose phosphates) by the aldolase. Dihydroxyacetone
phosphate (DHAP) is isomerized to glyceraldehyde 3-P, which is a
triose phosphate. The aldolase is named for the mechanism of the
forward reaction, which is an aldol cleavage, and the mechanism of
the reverse reaction, which is an aldol condensation.

Mechanism for the fructose-1,6 bisphosphate aldolase reaction
Never forget me!
The aldolase activates the substrate for cleavage by nucleophilic attack on the keto carbon at position 2 with a lysine -amino group in the
active site. This is facilitated by a protonation of the carbonyl oxygen by an active site (aspartate). The resulting carbinolamine undergoes
dehydration to give an iminium ion, or protonated Schiff base (Schiff base: imine; R2 . A Schiff base is a nucleophilic addition
product between an amin group and a carbonyl group.
A reto-aldol reaction then cleaves the protonated Schiff base into an enamine plus GAP. The enamine is
protonated to give another iminium ion (protonated Schiff base; reaction 4), which is then hydrolyzed off
the enzyme to give the second product, DHAP.
The Schiff base intermediate is advantageous in this reaction because it can delocalize electrons. The
positively charged iminium ion is thus a better electron acceptor than a ketone carbonyl, facilitating retro-
aldol reactions. This reaction also demonstrates why it was important to isomerize G6P to F6P in reaction
2. If glucose had not been isomerized to fructose (moving the carbonyl from C-1 to C-2), then the
aldolase reaction would have given two- and four-carbon fragments, instead of the metabolically
equivalent three-carbon fragments.
Energy Payoff Phase
Electron carrier
^ 2 ATP 2 ATP
2 NADH 2 H2O
Glyceraldehyde 2 ADP
3-phosphate (G3P) 2 NAD 2H 2 ADP 2 2 2 2
2
Triose Phospho- Phospho- Enolase Pyruvate

phosphate 2 Pi glycerokinase glyceromutase kinase
dehydrogenase
1,3-Bisphospho- 3-Phospho- 2-Phospho- Phosphoenol- Pyruvate
glycerate glycerate glycerate pyruvate (PEP)
In the energy payoff phase of the glycolytic pathway, glyceraldehyde 3-P is oxidized and phosphorylated so that subsequent
intermediates of glycolysis can donate phosphate to ADP to generate ATP. The first reaction in this sequence, catalyzed by
glyceraldehyde 3-P dehydrogenase (a Triose phosphate dehydrogenase), is really the key to the pathway. This enzyme
oxidizes the aldehyde group of glyceraldehyde 3-P to an enzyme-bound carboxyl group and transfers the electrons to NAD+
to form NADH. The oxidation step is dependent on a cysteine residue at the active site of the enzyme, which is followed by
the process of substrate-level phosphorylation (the formation of a high-energy phosphate bond where none previously
existed, without the use of oxygen).
In the next (7th) reaction, the phosphate in this bond is transferred to ADP to form ATP by phosphoglycerate kinase. The
energy of the acyl phosphate bond is high enough (~10 kcal/mol) so that transfer to ADP is an energetically favorable
process. Another product of this reaction is 3-phosphoglycerate.

Reactions of the Glyceraldehyde 3-P dehydrogenase
2 ATP 2 ATP
2 NADH 2 H2O
Glyceraldehyde 2 ADP
3-phosphate (G3P) 2 NAD 2H 2 ADP 2 2 2 2
2
Triose Phospho- Phospho- Enolase Pyruvate

phosphate 2 Pi glycerokinase glyceromutase kinase
dehydrogenase
1,3-Bisphospho- 3-Phospho- 2-Phospho- Phosphoenol- Pyruvate
glycerate glycerate glycerate pyruvate (PEP)
can be deprotonated depending The glyceraldehyde 3-P dehydrogenase glyceraldehyde (a Triose

on
pina value phosphate dehydrogenase) oxidizes the aldehyde group of glyceraldehyde
3-P to an enzyme-bound carboxyl group and transfers the electrons to NAD+
to form NADH. The oxidation step is dependent on a cysteine residue at the
active site of the enzyme, which forms a high-energy thioester bond during
the course of the reaction. The high-energy intermediate immediately
accepts an inorganic phosphate to form the high-energy acyl phosphate
bond in 1,3-bisphosphoglycerate (High-energy phosphates are indicated by
the red squiggles in the Figure), releasing the product from the cysteine
residue on the enzyme. This high-energy phosphate bond is the start of
substrate-level phosphorylation (the formation of a high-energy phosphate
bond where none previously existed, without the use of oxygen).

Reactions in steps 8-10 of the Energy Payoff Phase
2 ATP
2 H2O
2 ADP
2 2 2 2
Phospho- Enolase Pyruvate

glyceromutase kinase
3-Phospho- 2-Phospho- Phosphoenol- Pyruvate

glycerate glycerate pyruvate (PEP)
To transfer the remaining low-energy phosphoester on 3-phosphoglycerate to

ADP, it must be converted into a high-energy bond. This conversion is
accomplished by moving the phosphate to the second carbon (forming 2-
phosphoglycerate) and then removing water to form phosphoenolpyruvate
(PEP). The enolphosphate bond is a high-energy bond (its hydrolysis releases
approximately 14 kcal/mol of energy), so the transfer of phosphate to ADP by
pyruvate kinase is energetically favorable. This final reaction converts PEP to
pyruvate.

Phases of the glycolytic pathway
The glycolytic pathway, which cleaves
Energy Investment Phase 1 mol of glucose to 2 mol of the three-
carbon compound pyruvate, consists of a
Glucose preparative phase (energy investment
phase) and an ATP-generating phase
(energy payoff phase). In the initial
preparative phase of glycolysis, glucose
is phosphorylated twice by ATP and
2 ATP used 2 ADP 2P cleaved into two triose phosphates.
In the ATP-generating phase,
glyceraldehyde 3-phosphate
(glyceraldehyde 3-P) (a triose
phosphate) is oxidized by NAD+ and
phosphorylated using inorganic
Energy Payoff Phase phosphate. The high-energy phosphate
bond generated in this step is transferred
to ADP to form ATP. The remaining
4 ADP 4 P 4 ATP formed phosphate is also rearranged to form
another high-energy phosphate bond that
is transferred to ADP. Because 2 mol of
triose phosphate were formed, the yield
from the ATP-generating phase is 4 mol
2 NAD 4e 4H 2 NADH 2H of ATP and 2 mol of NADH. The result
is a net yield of 2 mol of ATP, 2 mol of
NADH, and 2 mol of pyruvate per mole
of glucose.
2 Pyruvate 2 H2O
Net
Glucose 2 Pyruvate 2 H2O
4 ATP formed 2 ATP used 2 ATP
2 NAD 4e 4H 2 NADH 2H
The critical reaction steps in glycolysis
Note the 3 large negative standard free-energy changes in the 10 reaction steps of glycolysis.
values are estimated from the approximate intracellular concentrations of glycolytic intermediates in rabbit skeletal muscle.
Remember:
The thermodynamically favored direction of a
reaction is determined by changes in both the
enthalpy (H) and the entropy (S). The free
energy, G = H - TS, takes both into account.
The criterion for a favorable process is that free
energy change G = H - S is negative
exergonic rather than positive
endergonic ; this is one statement of the
second law of thermodynamics.
D. L. Nelson, M. & M. Cox, Lehninger Principles of Biochemistry, 4th edition

7th edition
Coffee break
Major sites of regulation in the glycolytic pathway
stimulators ←
I
inhibitors
Hexokinase and phosphofructokinase-1 are

the major regulatory enzymes in skeletal
muscle. The activity of pyruvate
dehydrogenase in the mitochondrion
determines whether pyruvate is converted to
lactate or to acetyl coenzyme A (acetyl-CoA).
The regulation shown for pyruvate kinase
occurs only for the liver (L) isoenzyme.

Regulation of Phosphofructokinase-1
Phosphofructokinase-1 (PFK-1) is an allosteric enzyme that has a total of six binding sites:
two are for substrates (MgATP and fructose 6-P) and four are allosteric regulatory sites. The
allosteric regulatory sites occupy a different domain on the enzyme than the catalytic site. The
allosteric sites for PFK-1 include an inhibitory site for MgATP, an inhibitory site for citrate, an
allosteric activation site for AMP, and an allosteric activation site for fructose 2,6-
bisphosphate (fructose 2,6-bisP).
ATP binds to two different sites on the enzyme: the substrate-binding site and an allosteric
inhibitory site. Under physiologic conditions, the ATP concentration is usually high enough to
saturate the substrate-binding site and inhibit the enzyme by binding to the ATP allosteric site.
This effect of ATP is opposed by AMP, which binds to a separate allosteric activator site. For
most of the PFK-1 isoenzymes, the binding of AMP increases the affinity of the enzyme for
fructose 6-P.
At high [ATP], PFK-1 behaves cooperatively, and the plot of enzyme activity versus [fructose-6-phosphate]
is sigmoid. High [ATP] thus inhibits PFK- -6-phosphate.

R. H. Garrett & C. M. Grisham, Biochemistry, 4th edition
Regulation of Glucose content
Overview of the major pathways of glucose metabolism. Pathways for production of blood glucose are shown by dashed lines.
Acetyl-CoA, acetyl coenzyme A; DHAP, dihydroxyacetone phosphate; FA, fatty acids; OAA, oxaloacetate; PEP,
phosphoenolpyruvate; TCA, tricarboxylic acid; TG, triacylglycerols.

Hormonal regulation of glycolysis
The regulation of glycolysis by allosteric activation or

inhibition, or the phosphorylation/dephosphorylation of
rate-limiting enzymes, is short term that is, they
influence glucose consumption over periods of minutes or
hours. Superimposed on these moment-to-moment effects
are slower, and often more profound, hormonal
influences on the amount of enzyme protein synthesized.
These effects can result in 10-fold to 20-fold increases in
enzyme activity that typically occur over hours to days.
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. These changes reflect an
increase in gene transcription, resulting in increased
enzyme synthesis. High activity of these three enzymes
favors the conversion of glucose to pyruvate, a
characteristic of the wellfed state. Conversely, gene
transcription and 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.
R. Harvey & D. Ferrier, BIOCHEMISTRY 5th edition

Biosynthetic functions of glycolysis
Glycolysis, in addition to providing ATP, generates

precursors for biosynthetic pathways. Intermediates
of the pathway can be converted to ribose 5-
phosphate, the sugar incorporated into nucleotides
such as ATP. Other sugars, such as UDP-glucose,
mannose, and sialic acid, are also formed from
intermediates of glycolysis. Serine is synthesized
from 3-phosphoglycerate, and alanine from
pyruvate. The backbone of triacylglycerols,
glycerol 3-P, is derived from DHAP in the glycolytic
pathway.

Anaerobic glycolysis
When cells have a limited supply of oxygen (e.g., the

kidney medulla), or few or no mitochondria (e.g., the red
blood cell), or greatly increased demands for ATP (e.g.,
skeletal muscle during high-intensity exercise), they rely
on anaerobic glycolysis for generation of ATP. In
anaerobic glycolysis, lactate dehydrogenase oxidizes the
NADH generated from glycolysis by reducing pyruvate to
lactate. Because O2 is not required to reoxidize the
NADH, the pathway is referred to as anaerobic. The
energy yield from anaerobic glycolysis (2 mol ATP per
mole of glucose) is much lower than the yield from
aerobic oxidation. The lactate (lactic acid) is released into
the blood. Under pathologic conditions that cause
hypoxia, tissues may generate enough lactic acid to cause
lactic acidemia.

Oxidative Fates of Pyruvate and Nicotinamide Adenine Dinucleotide
The NADH produced from glycolysis must be continuously reoxidized back to NAD+ to
provide an electron acceptor for the glyceraldehyde 3-P dehydrogenase reaction and
prevent product inhibition. Without oxidation of this NADH, glycolysis cannot continue.
There are two alternate routes for oxidation of cytosolic NADH.
One route is aerobic, involving shuttles that transfer reducing equivalents across the
mitochondrial membrane and ultimately to the electron-transport chain and oxygen
(A).
The other route is anaerobic (without the use of oxygen). In anaerobic glycolysis,
NADH is reoxidized in the cytosol by lactate dehydrogenase (LDH), which reduces
pyruvate to lactate (B).
Lactate dehydrogenase reaction
When the oxidative capacity of a cell is limited (e.g., in the red blood cell, which has no mitochondria), the
pyruvate and NADH produced from glycolysis cannot be oxidized aerobically. The NADH is therefore
oxidized to NAD+ in the cytosol by reduction of pyruvate to lactate. The reduction of pyruvate is catalyzed by
lactate dehydrogenase, which forms the L-isomer of lactate at pH 7:
Note: The overall equilibrium of this reaction strongly favors lactate

formation, as shown by the large negative standard free-energy change.

Fermented milk products
Yogurt and Dahi are fermented milk products produced by bacterial fermentation of
milk. The bacteria, Lactobacillus and Streptocossus, used to make yogurt are known
as "yogurt cultures". Fermentation of lactose by these bacteria produces lactic acid,
which acts on milk proteins to give yogurt and dahi the texture and characteristic tang.
Fate of Lactate
Carl F. Cori (1897-1984)

Gerty T. Cori (1896-1957)
Lactate released from cells that undergo anaerobic glycolysis is taken up by other tissues (primarily the liver, heart,
and skeletal muscle) and oxidized back to pyruvate. In the liver, the pyruvate is used to synthesize glucose
(gluconeogenesis), which is returned to the blood. The cycling of lactate and glucose between peripheral tissues
and liver is called the Cori cycle.
The heart, with its huge mitochondrial content and oxidative capacity, is able to use lactate released from other
tissues as a fuel. During exercise such as bicycle riding, lactate released into the blood from skeletal muscles in the
leg might be used by resting skeletal muscles in the arm. In the brain, glial cells and astrocytes produce lactate,
which is used by neurons or released into the blood.

Voet, Voet, Pratt: Principles of BIOCHEMISTRY 3rd edition
Tissues of the eye are also partially dependent on anaerobic glycolysis
The eye contains cells that transmit or focus light,

and these cells, therefore, cannot be filled with
opaque structures such as mitochondria or densely
packed capillary beds. The corneal epithelium
generates most of its ATP aerobically from its few
mitochondria but still metabolizes some glucose
anaerobically. Oxygen is supplied by diffusion from
the air.
The lens of the eye is composed of fibers that must
remain birefringent to transmit and focus light, so
mitochondria are nearly absent. The small amount of
ATP required can be generated from anaerobic
glycolysis even though the energy yield is low. The
lens is able to pick up glucose and release lactate
into the vitreous body and aqueous humor. It does
not need oxygen and has no use for capillaries.

Anaerobic conversion of Pyruvate to Ethanol
The synthesis of ethanol by highly selected strains of yeast is important in the production of
beer and wine. Yeast cells convert pyruvate to ethanol and CO2 and oxidize NADH to
NAD+. Two reactions are required:
Pyruvate is decarboxylated in an irreversible reaction catalyzed by pyruvate
decarboxylase. This reaction is a simple decarboxylation and does not involve the net
oxidation of pyruvate. Pyruvate decarboxylase requires Mg2+ and has a tightly bound
coenzyme, thiamine pyrophosphate (TPP).
Alcoholdehydrogenase catalyzes the reduction of acetaldehyde to ethanol. This
oxidation-reduction reaction is coupled to oxidation of NADH.
The sum of the glycolytic reaction and the conversion of pyruvate to ethanol is
L. A. Moran, H. R. Horton, K. G. Scrimgeour & M. D. Perry, Principles in Biochemistry, 5th edition

TPP and its role in pyruvate decarboxylation
TPP is the coenzyme form of vitamin B1 (thiamine). The role of the functional group of thiamine
pyrophosphate (the reactive carbon in the thiazolium ring is shown in red) in formation of a covalent
intermediate. (A) A base on the enzyme (B) abstracts a proton from thiamine, creating a carbanion
(general acid base catalysis). (B) The carbanion is a strong nucleophile and attacks the partially
positively charged keto group on the substrate. (C) A covalent intermediate is formed, which, after
decarboxylation, is stabilized by resonance forms. The uncharged intermediate is the stabilized transition-
state complex.
L. A. Moran, H. R. Horton, K. G. Scrimgeour & M. D. Perry, Principles in Biochemistry, 5th edition

Production of Swiss Cheese
Three types of bacteria are used in the production of Emmental

cheese: Streptocossus salivarius, Lactobacillus, and
Propionibacterium. In a late stage of cheese production, the
propionibacteria consume the lactic acid excreted by the other
bacteria and release acetate, propionc acid, and carbon dioxide
gas. The carbon dioxide slowly forms the bubbles that develop
the "eyes". The acetate and propionic acid give Swiss its nutty
and sweet flavor.
In general, the larger the eyes in a Swiss cheese, the more
pronounced its flavor because a longer fermentation period gives
the bacteria more time to act.
http://en.wikipedia.org/wiki/Swiss_cheese
Fermentation Overview
© 2017 Pearson Education, Ltd

Major pathways for fermentation of sugars
Major pathways for fermentation of sugars including

organisms involved and end products formed
V. Müller (2001) Bacterial Fermentation. ENCYCLOPEDIA OF LIFE SCIENCES

Industrial products from fermentations
V. Müller (2001) Bacterial Fermentation. ENCYCLOPEDIA OF LIFE SCIENCES

The evolutionary significance of Glycolysis
Glycolysis is the most common metabolic pathway among organisms on Earth, indicating that it
evolved early in the history of life
Early prokaryotes may have generated ATP exclusively through glycolysis due to the low oxygen
content in the atmosphere
The location of glycolysis in the cytosol also indicates its ancient origins; eukaryotic cells with
mitochondria evolved much later than prokaryotic cells
CITRIC OXIDATIVE
GLYCOLYSIS PYRUVATE
ACID PHOSPHORYL-
OXIDATION
CYCLE ATION
ATP

Comparison of fermentation and respiration
Glucose
Glycolysis
CYTOSOL
Pyruvate
No O2 present: O2 present:
Fermentation Aerobic cellular
respiration
MITOCHONDRION
Ethanol, Acetyl CoA
lactate, or
other products
CITRIC
ACID
CYCLE

Quiz
one of the following statements concerning glycolysis is correct?

A. The conversion of glucose to lactate requires the presence of oxygen.
B. Hexokinase is important in hepatic glucose metabolism only in the absorptive period
following consumption of a carbohydrate-containing meal.
C. Fructose 2,6-bisphosphate is a potent inhibitor of phosphofructokinase.
D. The regulated reactions are also the irreversible reactions.
E. The conversion of glucose to lactate yields two ATP and two NADH.
2. The reaction catalyzed by phosphofructokinase-1:

A. is activated by high concentrations of ATP and citrate.
B. uses fructose 1-phosphate as substrate.
C. is the rate-limiting reaction of the glycolytic pathway.
D. is near equilibrium in most tissues.
E. is inhibited by fructose 2,6-bisphosphate.
Quiz
3. A major role of glycolysis is which of the following?

A. To synthesize glucose
B. To generate energy
C. To produce FAD(2H)
D. To synthesize glycogen
E. To use ATP to generate heat
Quiz
4. Which one of the following organs has the highest demand for glucose as a fuel?
A. Brain
B. Muscle (skeletal)
C. Heart
D. Liver
E. Pancreas
The brain requires glucose because fatty acids cannot readily cross the blood brain barrier to
enter neuronal cells. Thus, glucose production is maintained at an adequate level to allow the
brain to continue to burn glucose for its energy needs. The other organs listed as possible
answers can switch to the use of alternative fuel sources (lactate, fatty acids, amino acids) and
are not as dependent on glucose for their energy requirements as is the brain.
Thank you!

Glycolysis and Metabolic Fates of Pyruvate: Prof. Dr. Gerhard Grüber

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Glycolysis and Metabolic Fates of Pyruvate: Prof. Dr. Gerhard Grüber

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Glycolysis and Metabolic fates of pyruvate

Prof. Dr. Gerhard Grüber

More reduced state Less reduced state

Fats Carbohydrates Carbonyls Carboxyls Carbon

Garett & Grisham: Biochemistry 4th edition

A ball bouncing down a flight

Third shell (highest energy

Second shell (higher Energy

First shell (lowest

© 2017 Pearson Education, Ltd.

© 2017 Pearson Education, Ltd.

The major carbohydrates in the diet are starch,

glucose. Other dietary sugars (mainly fructose +

After glucose is transported into cells, it is

In tissues, fructose and galactose are

Overview of fructose and galactose metabolism.

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

can occur w/o oxygen

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

ATP Glucose Fructose ATP Fructose

Hexokinase Phosphogluco- Phospho- Aldolase

Never forget me!

ATP Glucose Fructose ATP Fructose

Hexokinase Phosphogluco- Phospho- Aldolase

conch still binds well nigh catalyst ng

© 2017 Pearson Education, Ltd.

J. M. Berg, J. L. Tymoczko & L. Stryer, Biochemistry 7th edition

ATP Glucose Fructose ATP Fructose

Hexokinase Phosphogluco- Phospho- Aldolase

© 2017 Pearson Education, Ltd.

ATP Glucose Fructose ATP Fructose

Hexokinase Phosphogluco- Phospho- Aldolase

Fructose 1,6-bisP is cleaved into two phosphorylated three-carbon

© 2017 Pearson Education, Ltd.

Never forget me!

Triose Phospho- Phospho- Enolase Pyruvate

© 2017 Pearson Education, Ltd.

Triose Phospho- Phospho- Enolase Pyruvate

can be deprotonated depending The glyceraldehyde 3-P dehydrogenase glyceraldehyde (a Triose

© 2017 Pearson Education, Ltd.

Phospho- Enolase Pyruvate

3-Phospho- 2-Phospho- Phosphoenol- Pyruvate

To transfer the remaining low-energy phosphoester on 3-phosphoglycerate to

© 2017 Pearson Education, Ltd.

D. L. Nelson, M. & M. Cox, Lehninger Principles of Biochemistry, 4th edition

Hexokinase and phosphofructokinase-1 are

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

The regulation of glycolysis by allosteric activation or

R. Harvey & D. Ferrier, BIOCHEMISTRY 5th edition

Glycolysis, in addition to providing ATP, generates

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

When cells have a limited supply of oxygen (e.g., the

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

Note: The overall equilibrium of this reaction strongly favors lactate

D. L. Nelson, M. & M. Cox, Lehninger Principles of Biochemistry, 4th edition

Carl F. Cori (1897-1984)

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

The eye contains cells that transmit or focus light,

C. M. Smith, A. D. Marks & M. A. Liebermann , 7th edition

L. A. Moran, H. R. Horton, K. G. Scrimgeour & M. D. Perry, Principles in Biochemistry, 5th edition

L. A. Moran, H. R. Horton, K. G. Scrimgeour & M. D. Perry, Principles in Biochemistry, 5th edition