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Cellular Respiration

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Cellular Respiration

A. Malcolm Campbell, PhD

Christopher J. Paradise, PhD
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Cellular Respiration
Copyright © A. Malcolm Campbell and Christopher J. Paradise. 2016.

All rights reserved. No part of this publication may be reproduced, stored

in a retrieval system, or transmitted in any form or by any means—
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brief quotations, not to exceed 250 words, without the prior permission
of the publisher.

First published in 2016 by

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 Abstract
What happens to a meal after it is eaten? Food consists primarily of
lipids, proteins and carbohydrates (sugars). How do cells in the body
process food once it is eaten and turned it into a form of energy that
other cells can use? This book examines some of the classic experimen-
tal data that revealed how cells break down food to extract the energy.
Metabolism of food is regulated so that energy extraction increases when
needed and slows down when not needed. This type of self-regulation
is all part of the complex web of enzymes that convert food into energy.
Adding to this complexity is that all food eventually winds up as two
carbon bits that are all processed the same way. This book will also reveal
why animals breathe oxygen and how that relates to the end of the en-
ergy extraction process and oxygen’s only role in the body. Rather than
look at all the details, this book takes a wider view and shows how cel-
lular respiration is self-regulating.

Keywords
first law of thermodynamics, second law of thermodynamics, entropy,
free energy, reduced, oxidized, homeostasis, palmitic acid, casein, lactose,
enymes, redox, cofactors, coenzymes, beta oxidation, acetyl CoA, mitro-
chondrial matrix, cellular respiration, limiting factor, deamination, alloste-
rically modulated, phosphofructokinase, citrate, negative feedback loop,
mitochondrial matrix, chemiosmosis, ATP synthase
 Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 Molecules Carry Energy in their Covalent Bonds...............1
Chapter 2 Converting Common Foods into Energy.........................11
Lipid Metabolism.............................................................13
Protein Metabolism..........................................................20
Carbohydrate Metabolism................................................25
Ethical, Legal, Social Implications:
Memorizing Details Obscures Learning........................32
Chapter 3 Energy Extraction from 2-Carbon Intermediates..............39
Chapter 4 ATP Production from Digested Foods.............................47
Overview of Energy Homeostasis.....................................54
Ethical, Legal, Social Implications:
The Importance of Eating Vegetables............................57
Conclusion............................................................................................61
Glossary................................................................................................63
Index....................................................................................................65
Free ebooks ==> [Link]

Preface
This book on cellular respiration is part of a thirty book series that col-
lectively surveys all of the major themes in biology. Rather than just pres-
ent information as a collection of facts, the reader is treated more like a
scientist, which means the data behind the major themes are presented.
Reading any of the thirty books by Campbell and Paradise provides read-
ers with biological context and comprehensive perspective so that readers
can learn important information from a single book with the potential to
see how the major themes span all size scales: molecular, cellular, organ-
ismal, population and ecologic systems. The major themes of biology en-
capsulate the entire discipline: information, evolution, cells, homeostasis
and emergent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
Readers will learn, in an engaging and very non-conventional way, how
cells extract energy from lipids, proteins and carbohydrates and some
of the supporting evidence behind our understanding. The historic and
more recent experiments and data will be explored. Instead of believing
or simply accepting information, readers of this book will learn about the
science behind cellular respiration the same way professional scientists
do—with experimentation and data analysis. In short, data are put back
into the teaching of biological sciences.
Readers of this book who wish to see the textbook version of this content
can go to [Link]/icb where they will find pedagogically-
designed and interactive Integrating Concepts in Biology for introductory
biology college courses or a high school AP Biology course.

[Link]
 Acknowledgments
Publishing this book would not have been possible without the generous
gift of Dr. David Botstein who shared some of his Breakthrough Prize
with AMC. David’s gift allowed us to hire talented artists (Tom Webster
and his staff at Lineworks, Inc.) and copyeditor Laura Loveall. Thanks go
to Kristen Mandava for project management and guidance on the pub-
lishing process. In particular, we are indebted to Katie Noble and Melissa
Hayban for their many hours of help and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to admin-
istrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond, Verna
Case, and Barbara Lom who had confidence in us and encouraged us to
persist despite setbacks along the way.
These books were the product of the shared labor of my two vision-
ary coauthors Laurie Heyer and Chris Paradise. We shared the dream and
the hardships and developed this book from scratch. My family has been
very supportive and I thank Susan, Celeste and Paulina for their support
and patience. I also want to thank Jan Serie, my pedagogical mentor, who
taught me so much about the art and science of helping students learn.
I benefited from the support of the Howard Hughes Medical Institute
grant 52006292, the James G. Martin Genomics Program, and Davidson
College. This book would not have survived its first draft without my
students who endured the typos and the early versions of this book. These
undergraduates participated in a bold experiment to see if beginners could
construct their own knowledge, retain what they learned, and transform
the way they see themselves and the discipline of biology. While many
people said that beginning students were not up to the task, my students
proved them wrong.
 Introduction
Living takes energy and energy comes from the food we eat. How is a
sandwich converted into the molecules that muscles use to chew the
sandwich? It is hard to comprehend many molecular processes, especially
energy extraction. Energy cannot be seen, tasted or smelled and yet its
properties are experienced all the time. This book reveal how molecules
in a person’s cells convert the eaten food into energy that can be used by
cells. This exploration begins with a review of some chemistry terms. It
is important to remember how molecular energy is stored in chemical
bonds. It is also important to remember the basics of how energy is mea-
sured and described by chemists. Chemistry is a foundational science that
helps make biology understandable, especially at the molecular level. This
book focus on how cellular metabolism gradually breaks bigger molecules
into smaller ones while transferring the energy from the bonds in food
to bonds in molecules used to transport energy between and within cells.
Ultimately, cells produce molecules of ATP which can be used for many
functions. Along the way, cellular metabolism produces waste products
containing carbon and oxygen atoms as well as wasted energy cells were
unable to harness.
 CHAPTER 1

Molecules Carry Energy

in their Covalent Bonds

Everyone has watched water boil, and perhaps over an open fire. From
personal experience it is clear that burning wood releases a lot of heat
energy, but the molecular source of this energy may not be clear, nor is
how the energy is released. Notice that with a campfire, some of the heat
is radiated to the air around the fire, and the remaining heat is absorbed
by the metal pot and eventually the water to produce steam. Convert-
ing liquid water into gaseous steam is work, as defined in the chemical
sense. Converting matter from one state into another is a type of work
that requires an input of energy, such as heat from the fire. It is possible
to harness steam to do other work, which is what happens inside coal
and nuclear power plants. Steam spins paddles of a turbine, which causes
a magnet inside coiled wires to spin. A spinning magnet inside coiled
wires produces a flow of electrons (e-), which is electricity. Wind genera-
tors and hydroelectric dams also produce a flow of electrons by spinning
magnets inside wire coils. Solar panels produce electron flow from un-
stable compounds that absorb the sun’s energy. In short, electrons are a
fundamental form of energy that are consumed every time an electrical
device is turned on.
There are two physical laws related to energy that apply to everything
in the universe. The field of study that focuses on the conversion of heat
to mechanical work is called thermodynamics, coming from the Greek
terms for heat (thermo) and power (dynamics). The field of thermody-
namics has two laws that are pertinent to understanding of molecular
energy sources used to do work. The first law of thermodynamics states
that energy can be transformed (changed from one form to another) but
2 CELLULAR RESPIRATION

cannot be created or destroyed. The second law of thermodynamics

states that the entropy, or disorder, of a system increases over time unless
energy is expended to reduce entropy. Biology has many hypotheses, a
few of which have been supported often enough that they are elevated to
the status of theories. In physics, theories that have been demonstrated to
be fundamental to all actions within the universe are called laws to signify
their fundamental roles in nature.
In addition to the boiling water, campfires produce smoke that con-
tains CO2, water vapor, ashes, and lost heat, all of which are byproducts
of releasing the energy from the wood. The match used to start the fire
supplied enough energy to break a few covalent bonds of the wood and
produce CO2 and H2O from oxygen and the wood. The new covalent
bonds in CO2 and H2O carry less energy than the covalent bonds in
wood. The energy released by the initial burning wood promotes the
breaking of more covalent bonds in the wood, and the fire spreads
while more CO2 and H2O are produced. Smoke, ash, and wasted heat
are examples of increased entropy, abbreviated S, as a consequence of
burning the wood. Chemists and physicists have quantified the intercon-
version of energy, heat, and work in three components:

• Total energy of the reaction = H (total release of heat from

the fire)
• Free energy available to do work = G (heat absorbed by pot
and water)
• Entropy or disorder = S (unused energy in smoke, ashes and
heated air)

When looking at the wood before it burned, there was a lot of poten-
tial energy. The burned wood contained much less energy than the
unburned wood. Converting unburned wood to burned wood released
the energy of the system (H). The change in H (ΔH = change in the heat
of the reaction; Greek letter delta (Δ) indicates change) value for burning
wood would be negative, meaning the ashes and smoke (H2O and CO2)
contain less H than the unburned wood did. It makes sense, therefore,
that the ΔG for burning wood is also negative, because ΔG is a subset of
ΔH. Using the first law of thermodynamics, all of the heat stored in wood
 Molecules Carry Energy in their Covalent Bonds 3

has to be somewhere, and this law of nature can be described quantitatively

as: ΔG = ΔH – ΔS × T which looks like a game to guess the word g-h-o-
s-t. This equation says that the change (Δ) in energy available to do work
(G = free energy used to boil water) is the difference between the change
in heat of the reaction (ΔH = change in the wood’s potential energy) and
the change in disorder (ΔS = ashes, smoke, and heat lost to the area) times
the temperature (T = air temperature before the fire started). Burning
wood produces ΔS with a positive value, but ΔG and ΔH are both nega-
tive values. Covalent bonds in wood that contain larger amounts of free
energy are less stable, and they break more easily than the more stable,
lower energy covalent bonds in CO2 and H2O. The air temperature in the
entire campsite is unchanged (due to its vast size), as is the air pressure, so
these components of the equation are constant while the fire is burning.
It might be difficult to picture where wood’s energy is located prior
to its burning. To help see the source of wood’s hidden energy, think of
wood’s molecular structure as a series of atoms held together by covalent
bonds (Figure 1). ATP is a high energy molecule that should be famil-
iar from previous studies in biology. ATP is a nucleotide and an energy
source, but where is the energy? Every covalent bond contains a pair of

covalent bond

phosphorous

nitrogen

oxygen

carbon
hydrogen

Figure 1  Covalent bonds contain high energy electrons. A molecule

of ATP with atoms depicted as spheres and covalent bonds depicted
as thick lines connecting atoms of carbon, oxygen, hydrogen, nitrogen
and phosphorous.
Source: Original art from ATP Jsmol.
4 CELLULAR RESPIRATION

shared electrons, and the bond that will be consumed in most ATP re-
actions is the bond that holds the last phosphate onto the penultimate
phosphate. Remember that electricity is moving electrons, and the elec-
trons in the covalent bonds of wood are also a source of energy. Wood
contains trillions of covalent bonds, and their energy is released as heat
when fire breaks the covalent bonds. Chapter 2 will expand on the release
of energy from covalent bonds. Until then, it is important to understand
how chemists can measure the amount of energy in bonds.
Chemists have devised two different pieces of equipment to measure the
heat of a reaction ΔH and the amount of energy available to do work ΔG
(Figure 2). The bomb calorimeter is a modification of the first machines
designed in the 1780s to measure the amount of heat energy given off from
breaking all the covalent bonds of a compound. The modern versions of
these devices are essentially unchanged and consist of five basic parts: a water
bath, a reaction chamber, a stirring rod, a thermometer, and a spark gen-
erator to initiate the chemical reaction. The reaction chamber is filled with
the compound of interest and oxygen gas to facilitate the burning. As the

thermometer
(measure = heat
stir of the reaction, H)
voltameter
spark 0
reactant 1 reactant 2
–V +V
e– e–
e– e–
water e– e–
R1ox R2ox
oxygen R1red R2red
compound + –
– +
+ – – + – + – +
A B

Figure 2  Measuring energy released from chemical bonds. A, Bomb

calorimeter burns a compound of interest (ΔH) inside a container
submerged in water. B, Electrochemical cell allows electrons to flow
to reduce reactant 2, while counter ions flow to oxidize reactant 1.
The device measures voltage, and you calculate ΔG from voltage.
Source: Original art.
Free ebooks ==> [Link]

Molecules Carry Energy in their Covalent Bonds 5

compound loses the energy in its covalent bonds due to burning, the heat is
absorbed by a known volume of water, and the increase in water temperature
is quantified. The heat is derived from the chemical breakdown of covalent
bonds the same way wood releases its energy when burned. The amount of
releasable heat in a compound, ΔH, is quantified by a bomb calorimeter.
Biologists tend to be more interested in the amount of free energy
available to do molecular work, ΔG (heat used to boil the water) rather
than the total released heat of the reaction, ΔH, which includes smoke,
ashes and wasted heat. To understand what ΔG is, it is important to
understand how ΔG is determined experimentally in an electrochemical
cell (Figure 2B). By tradition, the reaction container on the left is used
as a standard chemical reaction, such as the interconversion of oxidized
H+ ions and reduced H2 gas. Into the reaction container on the right
is place the compound that is being studied in reduced and oxidized
forms, such as the biologically significant molecules of oxidized nico-
tinamide adenine dinucleotide (NAD+) and reduced nicotinamide ad-
enine dinucleotide plus hydrogen (NADH). Remember from previous
chemistry lessons that reduction is the acquisition of electrons, often
in the form of new covalent bonds. Furthermore, the reaction takes
place in water (the “universal solvent”) because the molecular polarity
of its partially charged oxygen and hydrogen atoms that allow it to dis-
solve more substances than any other liquid. Oxidation is the loss of
electrons from broken covalent bonds. The standard reaction between
H+ and H2 can serve as an electron source to reduce NAD+ to NADH
in the right chamber of Figure 2B. Alternatively, the compound being
studied might produce an oxidizing reaction and generate electrons
that would reduce H+ to H2. Whichever way the electrons flow, the
voltameter quantifies the rate of electron flow as well as the direction
of the flow, left to right in Figure 2B. A reducing chemical reaction on
the right would produce a positive voltage, whereas an oxidizing reac-
tion would produce a negative voltage. Below the reaction chambers is a
tube that allows positive or negative ions to flow in opposite directions
to keep the overall charge neutral, or maintain chemical homeostasis. In
short, an electrochemical cell exhibits atomic homeostasis because as
one chamber accumulates electrons, it also gets rid of other charged ions
so that the overall chamber remains electrically balanced. The energy of

[Link]
6 CELLULAR RESPIRATION

the paired electrons in a covalent bond is transferred from the oxidized

reactant to the reduced reactant in the electrochemical cell.
From a biological standpoint, the most important aspect of an elec-
trochemical cell is that it experimentally quantifies the electron flow from
a compound of interest, such as NADH being oxidized to NAD+. The
flow of electrons is electricity and an indirect measure of the amount
of free energy available to perform molecular work, or ΔG. To calculate
the precise value for the change in free energy of a compound as it was
oxidized, ΔG, one plugs the measured voltage from the electrochemical
cell into this equation: ΔG = 2nFε; ε = measured voltage from electro-
chemical cell; F = known constant based on the charge of an electron,
and n = known number of electrons per mole of compound. Look at the
following example for the oxidation of NADH to NAD+:

ε = voltameter of the overall reaction would read: +1.55 volts

(volts = joules/coulomb)
n = number of electrons per reaction: 2 moles of e- for one
mole of reaction
F = 96,485 coulombs of energy for one mole e- (a constant value)

Look at the units in the equation and simplify the units for ΔG:

joules mole e- coulomb

G = 3 3
coulomb mole reaction mole e-

joules mole e- coulomb

G = 3 3
coulomb mole reaction mole e-

joules
G =
mole reaction

The change in free energy available to perform work, ΔG, quanti-

fies the loss of potential energy if the molecule is being oxidized (burn-
ing wood or consuming ATP), which produced a negative value for ΔG
(Figure 3). ATP is a key unit of energy used to maintain the metabolic
economy of cells and molecules. When ATP loses its terminal phosphate
by the breaking of a covalent bond, adenosine diphosphate (ADP) con-
tains less total potential energy (ΔH) and less free energy available to do
work (ΔG). Referring back to the equation ΔG = ΔH – ΔS × T that re-
lates ΔG to ΔH, some logical conclusions can be drawn. Remember that
 Molecules Carry Energy in their Covalent Bonds 7

ATP

ADP

+ Pi
G = –32 kJ/mole

Figure 3  Converting ATP to ADP releases energy.

The terminal phosphate (Pi) is removed from ATP and
produced ADP. The products have less energy due to
the two electrons lost in the broken bond.
Source: Original art.

the total energy of a system (H) is greater than the free energy available
to do work (G). Starting with one molecule of ATP and ending with two
molecules (ADP and inorganic phosphate, Pi) in addition to lost heat, the
randomness of the system has increased; ΔS is a positive value. However,
reversing the chemical reaction and investing energy to make ATP from
ADP and Pi, there would be a new covalent bond, increased values for ΔG
(>0) and ΔH (>0), but decreased value for ΔS (<0).
Like steam, shared electrons in covalent bonds are potential energy.
Electrons are negatively charged and thus repel each other. Once liber-
ated from the covalent bond, the electrons carry with them some of the
energy (ΔG = free energy), but other portions of the energy holding the
electrons in the covalent bond will be lost to the environment as heat
(part of the ΔH, or total heat of the reaction). Any time a larger molecule
(such as, ATP) is broken down into two smaller molecules (such as, ADP
and Pi), molecular randomness or disorder (ΔS = entropy) is increased.
When bigger molecules are broken into smaller molecules with increased
disorder, air near the reaction will absorb some vibrational energy, which
we detect as heat as when wood is burned. Therefore, products from
burning wood or consuming ATP have negative values for ΔH and ΔG,
8 CELLULAR RESPIRATION

Table 1  Common biological molecules used in cellular respiration,

and their ΔG values.
reaction ΔG (kJ/mole)
ATP → ADP + Pi −32
+ +
NADH + ½ O2 + H → NAD + H2O −218
½ O2 + H2 → H2O −236
+ -
pyruvate: H + 5/2O2 + C3H3O3 → 2H2O + 3CO2 −1145
FADH2 + ½ O2 → FAD + H2O −199
glucose: 6O2 + C6H12O6 → 6 H2O + 6CO2 −2884

but positive values for ΔS. The production of polymers would produce
positive ΔH and ΔG values but a negative ΔS value, because energy was
invested in new covalent bonds between atoms, which decreased the mo-
lecular disorder in the system. Table 1 gives the ΔG values for some com-
mon biological molecules to increase understanding of where potential
energy is stored.
Think of a covalent bond as a tether holding two atoms together to
understand why some covalent bonds carry more energy than others.
Would two stallions tied together and two mice tied together need teth-
ers of equal strength? Of course not, because antagonistic horses run-
ning away from each other would require a stronger tether compared to
a pair of angry mice. Similarly, some atoms have greater repulsive forces
and thus require stronger covalent bonds and thus electrons with more
energy, both free energy (G) and total energy (H). Bomb calorimeters
measure all of the energy in bonds, which is measured as a change of heat
in the water (ΔH). Electrochemical cells measure the power of electrons
released, which could be used to perform molecular work (ΔG). From
Table 1 it is clear that burning glucose in the presence of O2 to produce
CO2 + H2O released the largest amount of free energy electrons (largest
negative ΔG value), whereas converting ATP to ADP + Pi released the
least amount of free energy (smallest negative ΔG value).
This chapter was intended as a refresher of chemical and physical ter-
minology and the important concept of moving energy from one covalent
bond to another with lower ΔG and ΔH. Cells require energy to main-
tain homeostasis. Each chemical reaction loses some energy to entropy;
 Molecules Carry Energy in their Covalent Bonds 9

so without an investment of more energy, the breakdown of food to ATP

gradually reduces the amount of ΔG and ΔH at each step. The molecules
inside cells have evolved to harness the energy carried by the electrons
and store it for other uses. Chapter 2 presents the main concepts and
highlights of cellular respiration, but all of the steps that take place will
not be presented.

Bibliography
Alberty RA. Calculation of standard transformed Gibbs energies and
standard transformed enthalpies of biochemical reactants. Arch
Biochem Biophys 353(1):116–130, 1998.
Atkins P, de Paula J. Physical chemistry for the life sciences. New York
2006, Freeman.
Borsook H, Winegarden HM. On the free energy of glucose and of tripal-
mitin. Proc Natl Acad Sci 16(9):559–573, 1930.
Darling S. Computation of the free energy of a-ketoglutarate and
pyruvate from constants of the transamination process. Nature 160:
403–404, 1947.
Gibbs JW. A method of geometrical representation of the thermodynamic
properties of substances by means of surfaces. Trans Conn Acad Arts
Sci, 2:382–404, 1873.
Nelson D, Cox M. Lehninger principles of biochemistry. ed 3, New York,
2000, Worth Publishing.
 CHAPTER 2

Converting Common
Foods into Energy

From a person’s first meal to his or her last, people consume three major
sources of energy: lipids, proteins, and carbohydrates. Mammals instinc-
tively drink milk—the ideal food for a newborn (Figure 4). The most
abundant lipid in human milk is palmitic acid, a 16-carbon fatty acid
that comprises about 25% of all the lipids in milk. Casein is the most
abundant human milk protein and constitutes about 40% of all milk
proteins. Two hundred and twenty-five amino acids are polymerized to
make human casein, and every amino acid contains two carbon atoms
and one nitrogen atom in addition to the atoms in each amino acid
side chain. The most abundant milk sugar, lactose, is composed of 12
carbons. Lactose comprises about 10% of all the milk carbohydrates.
Whether drinking mother’s milk or eating steak and potatoes, a person’s
body must digest complex foods into much simpler compounds in order
for to convert the food into ATP energy. This chapter illustrates how
lipids, proteins, and sugars are broken down into smaller pieces that cells
can use to produce ATP.
Quantifying the exact ΔG in casein as shown in Figure 4 is impos-
sible, because it is not possible to see the structure of the different amino
acid side chains of casein. Nevertheless, it is known that each amino acid
consists of two covalently linked carbons in the constant portion and
therefore casein has at least 450 carbons. The potential energy is not in
the carbons but in the covalent bonds holding the atoms together. There-
fore, one molecule of casein contains much more energy than one mol-
ecule of palmitic acid, which contains more energy than the sugar, lactose.
To determine which category of molecule provides the most energy in
milk, it would be necessary to know how many copies of each molecule
12 CELLULAR RESPIRATION

225  OH 
casein
MKVLILACLVALALARETIESLSSSEESITEYKQKVEKVKHEDQQQGEDEHQDKIYPSFQPQPL
IYPFVEPIPYGFLPQNILPLAQPAVVLPVPQPEIMEVPKAKDTVYTKGRVMPVLKSPTIPFFDP
QIPKLTDLENLHLPLPLLQPLMQQVPQPIPQTLALPPQPLWSVPQPKVLPIPQQVVPYPQRAVP
VQALLLNQELLLNPTHQIYPVTQPLAPVHNPISV

OH
lactose palmitic acid
B C

Figure 4  These are the most abundant protein, carbohydrate,

and fatty acid found in human breast milk. A, Casein is the most
abundant human milk protein; composed of 225 amino acids, each of
which contains at least one nitrogen and two carbon atoms. B, The
12-carbon disaccharide, lactose, comprises about 10% of all sugars
in milk. C, The 16-carbon fatty acid, palmitic acid, comprises about
25% of all human milk lipids.
Source: All images are original art based on public domain information.

are present in a given volume of mother’s milk. Regardless of their rela-

tive abundance, it is possible to see that all three molecules contain many
covalent bonds, and therefore they each have large negative ΔG values
when they are broken down into smaller molecules.
It is good to remember that enzymes catalyze chemical reactions,
making the reactions more likely to happen. Enzymes are not consumed
as a part of chemical reactions where covalent bonds are broken or made.
Many enzymes reduce one molecule and oxidize another, a reaction
described by the term redox. It is possible to approximate the degree
of oxidation or reduction of a molecule by evaluating the ratio of car-
bon to hydrogen or carbon to oxygen. Carbon bound to two oxygen
atoms, CO2, is fully oxidized and contains very little potential energy to
do work (small negative ΔG). Conversely, CH4 is fully reduced and has
a very large negative ΔG. Some enzyme-catalyzed redox reactions require
small inorganic cofactors or larger organic coenzymes in addition to the
substrates, because the cofactors facilitate the movement of electrons dur-
ing the chemical reaction. Cofactors such as, Mg2+ and Ca2+ are usually
 Converting Common Foods into Energy 13

unaffected by the chemical reaction, whereas coenzymes such as NAD+

(nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleo-
tide) can be covalently modified temporarily as part of the enzymatic re-
action. Enzymes position the reactants together in close proximity, which
significantly enhances the rate of chemical reactions that happen many
times per second.

Lipid Metabolism
In 1928, biologists were not sure how foods such as, lipids, proteins,
and carbohydrates were converted to fuel for cells. Armand Quick from
Cornell University in New York wanted to quantify what happens to
lipids of different lengths once they were ingested. Quick chose dogs as
his model system because earlier biochemical research on lipid metabo-
lism had also used dogs, and Quick wanted to build on the findings of
others. He synthesized a series of six lipids, all of which contained the
hexagonal benzene ring on the end opposite the acid (COOH) portion of
the fatty acid (Figure 5). These benzene-lipids are not natural products, so
the dogs could not digest the benzene portion of the fatty acid.
Other biochemists had hypothesized that lipids were broken into
smaller bits starting at the acid (COOH) side. Quick hypothesized that
if he synthesized non-biological lipids and fed them to dogs, he should
be able to collect the synthetic fatty acid waste products and determine
how much of the fatty acid was still attached to the benzene ring. No
matter how long the fatty acid was that Quick fed to the dogs, all of
the waste products were urinated as either benzoic acid or phenylacetic
acid (see Figure 5). When Quick covalently attached 16-carbon palmitic
acid to a benzene ring, it was secreted as phenylacetic acid. When he fed
the dogs either 15- or 17-carbon fatty acids, they were always secreted as
benzoic acid. He quantified the amount of benzoic acid and phenylace-
tic acid and verified that all of the synthetic fatty acid consumed by the
dogs had been accounted for in their urine samples as either benzoic or
phenylacetic acid.
When Quick synthesized the benzene-containing lipids, he made two
categories of fatty acids: those with an even number of carbons, and those
with an odd number of carbons. Within each of these categories, he had
14 CELLULAR RESPIRATION

O
OH
OH
O

benzioc acid phenylacetic acid

O
OH
OH
O

phenylproprionic acid phenylbutyric acid

O
OH
OH
O

cinnamic acid phenylisocrotonic acid

Figure 5  Determining how fatty acids are oxidized. Two

experimentally synthesized categories of fatty acids that share a
common 6-carbon ring structure referred to as a benzene ring or a
phenol group.
Source: All panels from public domain of chemical structures. Original art.

some with double bonds at various places—as seen in cinnamic acid and
phenyl isocrontonic acid (Figure 5). Quick wondered whether the num-
ber of double bonds in a fatty acid would affect the way lipids were broken
down into smaller bits. No matter how many double bonds were within
the fatty acid, those with an odd number of carbons always produced
benzoic acid, and those with an even number always produced phen-
ylacetic acid. From these results, Quick verified earlier predictions that
fatty acids are cut into smaller pieces, two carbons at a time, starting at
the end containing the acid group of COOH. As his experimental lipid
got smaller, it either terminated in one carbon plus benzene, or two car-
bons plus benzene, which dogs could not metabolize further; they urinate
the indigestible waste. The process of breaking fatty acids into 2-carbon
bits is called beta oxidation, because the second carbon from the acid
 Converting Common Foods into Energy 15

end is called the beta carbon. Just as dogs were unable to digest benzoic
and phenylacetic acid, humans cannot digest Olestra because its shape is
unnatural, and we did not evolve an enzyme that can perform beta oxida-
tion on this octopus-like lipid. Olestra contains many, many calories, but
humans cannot digest it which is why some food manufacturers used it to
produce low calorie potato chips. Undigested Olestra becomes part of a
person’s feces, which explains why 30% of all people suffer from diarrhea
after consuming Olestra. Food containing Olestra used to carry a warn-
ing label, but starting in 2003, the warning was been removed from food
labels but the fecal problems were not removed.
Once biochemists had determined that all fatty acids were oxidized
into 2-carbon fragments, they wanted to understand how (Figure 6).

ATP octanoic oxygen octanoic acid

added? acid? consumtion consumtion
yes no 25.1 moles 0.0 moles
yes yes 52.5 moles 24.3 moles

octanoic acid
liver extract
no no 26.3 moles 0.0 moles
no yes 12.7 moles 0.0 moles

ATP
A

200 + + +
oxygen uptake (mm3)

100
+ – +

+ + –
0 – + +
0 10 20 30
B time (min)

Figure 6  Biochemical requirements for beta oxidation of an 8-carbon

lipid, octanoic acid. A, Liver cell extract was incubated with ATP
and octanoic acid as indicated for 30 minutes before oxygen and fatty
acid consumption were measured. B, Rate of oxygen consumption by
liver cell extracts over 30 minutes when octanoic acid and ATP were
added as indicated.
Source: A. Modified from Lehninger, 1945 his table 1. B. Modified from Lehninger, 1945, his figure 1.
16 CELLULAR RESPIRATION

By 1945, investigators knew that enzymes inside of cells were responsible

for beta oxidation, but they did not know how cells performed the fatty
acid digestion. To better understand the biochemistry, Albert Lehninger
working at The Johns Hopkins University in Baltimore, Maryland,
ground up liver cells to burst them open and release their cytoplasmic
contents. Lehninger incubated the liver cell extracts with many differ-
ent combinations of biological molecules, some of which are shown in
Figure 6A. With each combination, Lehninger measured consumption
of oxygen and the substrate octanoic acid. In a second set of experi-
ments, he measured the rate of oxygen consumption as a function of time
and the amount of each beta oxidation reactant. The experiments in
Figure 6B were the first time anyone had biologically digested fatty acids
without using live cells.
Lehninger found that ATP, oxygen, and fatty acids were consumed
during beta oxidation. Of course, cell extracts can do more than just beta
oxidation, which is why they consumed oxygen in all four conditions in
Figure 6. Only when ATP, oxygen, and octanoic acid were combined with
the cytoplasm of liver cells could Lehninger measure the consumption of
fatty acid and oxygen, which explains why fatty acid degradation is called
beta oxidation. By adding extra fatty acid, the liver cell extract consumed
more oxygen. When Lehninger added extra octanoic acid, the cells were
able to oxidize the fatty acid faster, and therefore they consumed oxygen
at a faster rate (Figure 6B).
The requirement for ATP was a major insight from these experi-
ments on beta oxidation (Figure 7). From his results, Lehninger cor-
rectly predicted that all fatty acids are oxidized into 2-carbon fragments
by protein enzymes. Fatty acids bind to enzymes, which use ATP to co-
valently link coenzyme A (CoA) onto the acid side of fatty acids. Pairs
of carbons are enzymatically cut off of the longer fatty acid to produce a
fatty acid shortened by two carbons plus acetyl-CoA; acetyl groups con-
tain two carbons. In addition, each round of beta oxidation produces
one FADH2 and one NADH, which are the reduced forms of FAD
and NAD+. Reduction indicates the formation of new covalent bonds,
which is the storage mechanism of potential energy transferred from the
fatty acid to the coenzymes. Liver cells also needed CoA, without which
enzymatic beta oxidation is impossible. Lehninger did not realize that
 Converting Common Foods into Energy 17

input = 1 palmitic acid + 8 CoA + 8 FAD + 8 NAD+ + 8 ATP

palmitic acid O

OH
coenzyme A

8th 1st
coenzyme A
2 carbon
acetyl
8  1 FADH2 group
8  1 NADH
output = 8 acetyl-CoA + 8 FADH2 + 8 NADH + 8 ADP acetyl-CoA

Figure 7  Degradation pathway for palmitic acid. Coenzyme A

(CoA) binds to palmitic acid before two carbons are removed from
the 16-carbon fatty acid. The process is repeated seven more times.
NAD+ is reduced to NADH, and FAD is reduced to FADH2 as
palmitic acid is oxidized.
Source: All art is original, based on common knowledge or PDB structures.

beta oxidation happens only inside of the mitochondrial matrix and

that lipid substrates must be imported into the organelle. To import
fatty acids into the mitochondria, CoA must have been covalently at-
tached to the fatty acid, which requires ATP. Once the CoA + fatty acid
reaches the mitochondrial matrix, it will be fully oxidized into as many
acetyl-CoA molecules as possible, which is determined by the number
of carbons in the fatty acid.
A search of the NCBI database called OMIM for beta oxidation
genetic diseases will return four different mutant alleles from different
genes that can inhibit fatty acid oxidation (Figure 8). This finding indi-
cates there are four different enzyme proteins are responsible for beta oxi-
dation, just as Lehninger had deduced. Different families exhibit distinct
but related symptoms caused by the inability to metabolize fatty acids of
particular length as measure by the number of carbons. When reading
the OMIM entries in detail, one would find some of the symptoms were
more acute when the patient fasted for some period of time, which could
be as simple as going longer between meals. Fatty acids are a good source
of energy that are often consumed after sugars have already been metabo-
lized. Notice that the OMIM data indicate the four different enzymes
metabolize fatty acids of particular lengths. When the structure of a very
18 CELLULAR RESPIRATION

lipids tested 16 carbons 14 carbons 8 carbons 4 carbons

family 1 high activity high activity low activity high activity
family 2 low activity high activity high activity high activity
family 3 high activity high activity high activity low activity
family 4 high activity low activity high activity high activity
A

Figure 8  Genetic basis for lipid metabolism diseases. A, Compiled data

summarizing the genetic capacity of four different families to oxidize
fatty acids of different lengths. B, Family pedigree and DNA sequence
analysis for VLCAD lipid metabolism disease gene. Circles are female
and squares are male. Filled shapes have the disease phenotype and
half filled shapes are heterozygotes.
Source: Panel A is composite data from OMIM, open access. Panel B from Strauss et al.,
1995; modified from their figure 4. Strauss, Arnold, Cynthia K. Powell, et al. 1995. Molecular
basis of human mitochondrial very-long-chain acyl-CoA dehydrogenase deficiency causing
cardiomyopathy and sudden death in children. PNAS. Vol. 92: 10496–10500. Copyright (1995)
National Academy of Sciences, U.S.A.

long chain acetyl-CoA dehydrogenase (VLCAD) enzyme is examined,

it becomes clear why there are four different genes are needed to oxidize
different length categories of fatty acids.
Figure 8B, shows a pedigree and the same short segment of DNA for
a family that has a mutation in their VLCAD gene. To obtain this gene
sequence, each family member donated some DNA. The genomic DNA
was sequenced, including introns and exons. The mutation in this par-
ticular family did not affect any exons, but there was a single base change
in the first base of intron number eleven. The dots and arrows on the data
help focus attention on the critical nucleotides where only the affected
male is homozygous for the mutation. The data were produced before
 Converting Common Foods into Energy 19

automated sequencing became popular, so each base has its own lane and
the sequence is read from bottom to top. For example, the first three bases
in all four samples are 59 TAA 39. The family depicted in Figure 8B suffers
from a recessive disease. It is recessive because only the homozygous male
has the disease. He has a G → A mutation in both alleles, and the rest
of his family is heterozygous at this position of intron 11. The mutation
blocks messenger RNA (mRNA) splicing and therefore an entire exon is
omitted from his mRNA and thus the protein is nonfunctional as would
be expected for a recessive disease.
From the structure of VLCAD, biologists discovered that the hydro-
phobic tail, opposite from where the CoA attaches to the acid (COOH),
extends into the binding cleft of the enzyme. Once in position, the
protruding CoA plus the first two carbons, alpha and beta carbons,
are cleaved from the rest of the fatty acid. Breaking the covalent bond
between the beta and gamma carbon produces a new acetyl-CoA and a
new acid group, COOH, on the newly shortened fatty acid’s freshly cut
end. A new CoA will be covalently linked (using ATP) to the COOH
end of the fatty acid before two more carbons can be removed to pro-
duce another acetyl-CoA product. From the OMIM data in Figure 8A,
it can be deduced that each length of fatty acid must fit into an appro-
priate sized enzyme that has an adequately deep binding pocket. The
binding pocket must be deep enough to hold the rest of the fatty acid
but not too deep. Once palmitic acid has been oxidized a couple times
by VLCAD, the shortened fatty acid must bind to a new enzyme, long
chain acetyl-CoA dehydrogenase (LCAD), for further oxidation. Based
on Figure 8A, it is possible to see that there are four sizes of dehydro-
genases that gradually oxidize the fatty acids. The production of acetyl-
CoA from fatty acids continues until all the carbons are attached to
CoA, or there is only one carbon remaining (see Figures 5 & 7).
If someone eats a lot of foods high in lipids, they will probably store
the excess carbon in the form of fat. The default storage of consumed fat
tells us that lipids are not oxidized immediately, and they can be stored
for use later. Beta oxidation, like all aspects of cellular respiration, is
regulated as part of molecular homeostasis. All organisms need energy
from cellular respiration to maintain their homeostasis, and the extrac-
tion of energy from food is regulated by homeostasis. Because lipids are
 Free ebooks ==> [Link]

20 CELLULAR RESPIRATION

fully oxidized once they enter the mitochondria, regulation of lipid me-
tabolism in eukaryotes must come before the lipids enter the mitochon-
dria. The limiting factor in metabolism of lipids is the abundance of
CoA. When cells have plenty of acetyl-CoA, any new lipid is stored as
fat. When cells have abundant CoA not bound to the 2-carbon acetyl
groups, then fatty acids (such as, palmitic acid in mother’s milk) is cova-
lently linked to CoA and immediately imported into the mitochondria
and fully oxidized to produce acetyl-CoA. Beta oxidation is regulated by
the availability of CoA as a limiting factor to regulate fatty acid diges-
tion versus storage. Limiting factors are familiar in everyday life because
money is an example that requires a homeostatic balance. No one can buy
more items than they have money to give. When people spend all of their
money, they have to stop buying. When they accumulate more money,
they can resume spending. Similarly, CoA is a limiting factor for lipid
metabolism. If the cell consumes all of its CoA and produces abundant
acetyl-CoA, then fatty acids cannot be imported into the mitochondria
for more beta oxidation. If CoA accumulates, mitochondria can resume
importing fatty acids and beta oxidation by using ATP to covalently
attach the lipid to CoA.
Lipids are oxidized into 2-carbon bits to produce FADH2, NADH,
and acetyl-CoA. Beta oxidation is regulated by CoA as a limiting fac-
tor. Cellular respiration can produce ATP from fatty acids, but ATP
production comes at the end of cellular respiration and is presented in
Chapter 4. Next, this chapter will present how proteins are broken down.
High-protein fad diets can produce short-term weight loss but there is a
negative consequence that makes high-protein diets risky.

Protein Metabolism
A healthy diet must include protein or else a person will suffer from mal-
nutrition. Luckily, a person’s first meal of mother’s milk contains many
proteins, including casein as the most abundant milk protein. Proteins
are composed of amino acids connected by covalent bonds called peptide
bonds because they are part of a protein or polypeptide. A whole protein
is too large to be broken down directly by cellular respiration enzymes,
so the first thing to do with a protein is depolymerize the amino acids

[Link]
 Converting Common Foods into Energy 21

into their monomers of amino acids. When food is eaten, it first reaches
the stomach, which is very acidic with a pH value of about 1 or 2. Very
low pH alone can break some peptide bonds, but this process is slower
than enzymatic cutting of peptide bonds. A pH of 1 or 2 is not a typical
environment for enzyme proteins to function, which presents a problem.
How can an enzyme digest proteins when the pH is so low that most
proteins cannot function?
Anyone who eats a lot of beef may be familiar with meat tenderizers
that can be used to make tough meat less chewy. The most common
meat tenderizer is papain, which is extracted from the papaya fruit and
is a protease that cleaves peptide bonds between amino acids. Papain
functions best at a pH around 6.5. To digest casein and other proteins
eaten, stomach cells secrete a protease that can function at very low pH
(Figure 9). Mammals produce many different proteases but the three
best known are trypsin, chymotrypsin, and pepsin. Each protease has an
optimal pH where the amino acids cause the protein to adopt a particu-
lar three-dimensional (3D) shape that maximizes the enzyme’s ability
to break peptide bonds in other proteins. It might be surprising to see
what happened when a variety of protein substrates were incubated with
pepsin. Figure 9B makes it clear why the human genome encodes several
different proteases.
The data in Figure 9A are from classic biochemistry experiments. Each
protease was mixed with a suitable substrate and a buffer of known pH.
After a fixed amount of time, the relative amount of enzyme activity was
quantified and graphed for each pH. In this comparison, it is clear why
pepsin is the primary protease that functions in the stomach. In fact, when
humans eat, stomach cells increase pepsin secretion to aid in protein diges-
tion. Most people are surprised that each protease cleaves some protein
substrates better than others, which helps explain why we have at least
two proteases that work in the small intestines (Figure 9A). Hemoglobin,
found in red meats, contains beneficial iron, and pepsin readily digests
the large protein into smaller fragments. Ovalbumin, also known as egg
white, is poorly digested by pepsin and requires additional proteolysis by
trypsin, chymotrypsin, and other enzymes found in small intestine.
Once the proteases have digested complex proteins into individual
amino acids, the molecular components of cellular respiration take
22 CELLULAR RESPIRATION

100 pepsin chymotripsin

trypsin

percent activity 50

0
1 2 3 4 5 6 7 8 9 10 11
A pH

0.7
hemogloblin
0.6
product formation

0.5 globin
0.4
0.3
0.2 ovalbumin
0.1
0.0
0 30 60 90 120 150 180 210 240
B time (min)

Figure 9  Protease activity as a function of pH. A, Humans produce

all three proteases tested, but each enzyme has a different pH
sensitivity. B, Pepsin was incubated with three different protein
substrates, and their degradation was monitored over 4 hours.
Source: A. Compiled from different sources and then standardized by percent activity. Original
art. B. Modified from Walker and Taylor, 1978; figure 2a. Reproduced with permission, Walker,
Valerie, and William H. Taylor. 1978. Ovalbumin digestion by human pepsins 1, 3 and 5.
Biochemical Journal. Vol. 176: 429–432. © the Biochemical Society.

control of converting amino acids into forms that are more easily con-
verted to ATP (Figure 10). One way to use amino acids that are con-
sumed is to recycle them by making new proteins directly from these
amino acids. However, if energy is needed from a protein food source
such as mother’s milk, cells have the ability to break amino acids into
two parts. Glutamate is one of the 20 amino acids and glutamate de-
hydrogenase (GDH) is an important enzyme in cellular respiration of
proteins. GDH consumes one amino acid and one NAD+ to produce an
ammonia molecule, one NADH, and the oxidized amino acid lacking its
 Converting Common Foods into Energy 23

amino
acid O O
H2N GDH O
OH e OH

R + NAD+ R + NADH
A + H2O + NH3

4.0
0.5 mM ADP 100 100

percent GDH activity

ratio GDH activity

3.0

2.0
50 50

1.0
0.5 mM ATP
0 0
0 100 200 300 0 1 2 0 50 100 150
NADH concentration (m M) C GTP (m M) ATP (mM)
B

Figure 10  Deamination of amino acids. A, GDH removes the amine

and produces ammonia and NADH as well as a 2-carbon acetyl
group. The circle labeled “e” represents an enzymatic reaction. B,
GDH activity is modulated by ATP and ADP over a range of NADH
concentrations. The ratio of activity compares GDH with and without
the indicated nucleotide. C, GTP and ATP alter GDH activity
differently at 85 µM NADH.
Source: A. Original Art. Panel B modified from Friedan, 1959; figure 2. Frieden, Carl. 1959.
Glutamic dehydrogenase: The effects of various nucleotides on the association-dissociation
and kinetic properties. Journal of Biological Chemistry. Vol. 234(4): 815–820. Copyright
1959 The American Society for Biochemistry and Molecular Biology. Panel C modified from
Frieden, 1963; his figure 7. Frieden, Carl. 1959. Glutamic dehydrogenase: The effects of
various nucleotides on the association-dissociation and kinetic properties. Journal of Biological
Chemistry. Vol. 234(4): 815–820. Copyright 1963 The American Society for Biochemistry and
Molecular Biology.

nitrogen atom. The R group in Figure 10A represents the side chain that
distinguishes the 20 amino acids from each other; different enzymes can
recognize particular R groups. For cellular respiration, the critical aspect
is not which amino acid is present, but that enzymes remove the nitrogen-
containing amino group (NH2) from amino acids. The removal of the
amino portion produces a 2-carbon acetyl acid connected to the R side
chain. Removal of an amine group is called deamination and takes
place inside mitochondria. Once again, cellular respiration oxidized a
complex molecule into 2-carbon bits in the mitochondria just as beta
oxidation did for fatty acids. Because deamination takes place in the mito­
chondria, the same reactions happen in both plants and animals since this
organelle is in both types of organisms.
24 CELLULAR RESPIRATION

Water is essential for life, some of its many functions may be unfamil-
iar to the reader (Figure 10A). One function of water is to facilitate the
deamination of amino acids. Water's oxygen is covalently attached to the
carbon that used to be connected to the nitrogen. One of the hydrogen
ions, H+, is added to NAD+, whereas the second hydrogen ion is found
in ammonia NH3, which used to be an amine group of NH2. When H+
covalently binds to NAD+, the two electrons (e-) in the new covalent
bond are used to neutralize the two positive charges in H+ and NAD+, so
the reduced molecule NADH is electrically neutral. All of the electrons
are accounted for, as are the carbons, nitrogen, and hydrogen atoms. The
formation of NADH from NAD+ and water produces a large positive
ΔG of +220 kJ/mole. The free energy in NADH will be extracted later
in cellular respiration (Chapter 4). Once ammonia is formed, the body
needs to excrete it. Ammonia is converted to urea and filtered from the
blood in the kidneys. Excessive protein diets can lead to high levels of am-
monia and damage the kidneys, which is one complicating side effect of
high protein diets. Urine contains urea, and its chemical formula NH2–
CO–NH2 indicates that each urea carries the nitrogenous waste of two
deaminated amino acids.
Beta oxidation of lipids was regulated by a limiting factor—CoA.
Deamination of amino acids is regulated differently (Figures 10B and 10C).
To regulate a biochemical pathway, the enzymes that accelerate the overall
chemical reactions need to be regulated. Enzyme activity can be modu-
lated by allosteric interactions with other molecules. ATP and GTP are
familiar molecules because these are two ribonucleic acids used in RNA
synthesis as well as energy sources. ATP is produced by the phosphoryla-
tion of ADP, which is the waste product when ATP is used as an energy
source. As seen in beta oxidation of fatty acids and deamination of amino
acids, NAD+ is reduced to NADH by cellular respiration and contains
a new covalent bond compared to the oxidized substrate of NAD+. The
data in Figure 10 provide a good understanding of the homeostatic regu-
lation of deamination.
Figures 10B and 10C indicate that deamination by GDH is allo-
sterically modulated by at least four molecules: ATP, GTP, ADP, and
NADH. ATP and GTP are both sources of readily usable energy that
inhibit deamination by GDH. GTP is over 100 times more potent at
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Converting Common Foods into Energy 25

inhibiting GDH than ATP. When ADP accumulates and molecular

energy stores are low, GDH is allosterically activated to produce more
NADH and 2-carbon acetyl groups. NADH, a source of potential energy,
also reduces GDH activity—as seen from the right portion of Figure 10A
where the ADP line levels off at high NADH levels. Therefore, deamina-
tion of amino acids is tightly coupled to the overall energy status of the
cell, which reveals another example of molecular homeostasis of energy
production. Without the regulation of GDH and other deaminating en-
zymes, all amino acids would be converted to 2-carbon acetyl groups,
stress the kidneys, and lose the opportunity to recycle amino acids directly
into new proteins. In short, homeostasis of deamination is adaptive by
supplying the cell with energy when really needed, but not producing
toxic waste when energy is plentiful from lipids or carbohydrates. Re-
member, mitochondria have their own genomes that are transcribed into
mRNA and translated into proteins all within the mitochondria. If all
amino acids were deaminated immediately upon arrival in the mitochon-
dria, the mitochondria could not translate the proteins encoded in its
genome, which would be fatal.

Carbohydrate Metabolism
Cellular respiration is a fundamental capacity shared by nearly every
species. Every organism and every cell relies upon energy to maintain
homeostasis of all cellular processes. Homeostasis also regulates the ex-
traction of energy from food sources, such as lipids and proteins. The
final category of food to consider is carbohydrate and in particular the
milk sugar, lactose.
Perhaps it is fitting to save carbohydrates for last, like a sweet dessert.
Carbohydrates include sugars and polymers of sugars, such as starch.
Carbohydrates are consumed when eating potatoes, pasta, or bread, or
drinking lactose in milk. As children, most people can digest lactose but
many adults lose the ability to produce the lactose-digesting enzyme,
lactase. Many people are lactose intolerant because they have stopped
making the sugar-digesting enzyme lactase that they used to make as
a child. Infants produced enough lactase in their intestines to break the
12-carbon lactose into two, slightly different 6-carbon sugars—glucose

[Link]
26 CELLULAR RESPIRATION

and galactose. Most mammals also produce an enzyme that converts ga-
lactose to glucose, so the focus the remaining portion carbohydrate me-
tabolism will be on digesting glucose. People whose ancestors have lived
in northern Europe, parts of Africa, and other locations where cow’s milk
has been consumed for many generations have mutations that permit
lifelong lactase production. Lactose intolerant people have retained the
wild-type allele that limits lactase production to childhood, whereas adult
milk drinkers carry the more recently evolved, mutated allele.
The enzymatic oxidation of 6-carbon glucose to a pair of the
3-carbon pyruvate is known as glycolysis (Figure 11). Note that energy in
the form of ATP is consumed during the first half of glycolysis and pro-
duced during the second half. The final step of glycolysis is the produc-
tion of two pyruvate molecules. One additional enzyme further oxidizes
both pyruvates by removing one CO2 and produces an acetyl group plus
one NADH from each of the two pyruvates. As with protein and lipid
metabolism, cellular respiration of glucose produces a 2-carbon acetyl
molecule as well as NADH as an electron carrier. The 2-carbon acetyl
group containing the remaining carbons from pyruvate is covalently at-
tached to CoA (the same as in beta oxidation of fatty acids). Acetyl-CoA
contains potential energy in the same way that NADH does by virtue
of its covalent bonds. It is not important to know the names of the
enzymes or the sugars between glucose and pyruvate, but notice that
two enzymes have been identified by name because of their important
homeostatic role in glycolysis. Unlike beta oxidation and deamination,
glycolysis happens in the cytoplasm of cells. In many species, including
humans, pyruvate can be fermented into lactic acid and CO2 whenever
insufficient oxygen is available. Lactic acid is why muscles burn during
extreme exercise.
Rather than focus on specific details of glycolysis, focus on what goes
in and what comes out of the pathway in Figure 11 (Table 2). For every
one glucose that enters glycolysis and finishes at pyruvate, the cell has a
net gain of two ATPs, two NADHs, and two pyruvates. The next oxida-
tive step, which happens inside the mitochondria of eukaryotes, produces
two more NADHs, a pair of CO2 molecules, and two acetyl-CoAs. Be-
cause the disaccharide lactose contains two six-carbon sugars, all of these
 Converting Common Foods into Energy 27

glucose

ATP + e

= carbon atom
= phosphate (PO43– ) + ADP

ATP + e phosphofructokinise

+ ADP

2 +
2 NAD+ + e

+ 2 NADH

2 ADP + e

+ 2 ATP

e
2 ADP + e pyruvate kinase

+ 2 ATP
2 pyruvates

2 NAD+ + e
coenzyme A
2 coenzyme A

+ 2 CO2 2 carbon
acetyl
group
+ 2 aceytl-CoA
+ 2 NADH

Figure 11  Glycolysis is the enzymatic conversion of glucose to

pyruvate. The 6-carbon sugar glucose is oxidized into two copies of
the 3-carbon pyruvate. Pyruvate is further oxidized to CO2 and an
acetyl group, which is covalently attached to CoA. Circles labeled “e”
represent enzymatic reactions. Substrates appear on the left of the
pathway and products on the right. CO2 is a waste product.
Source: Original art based on common knowledge and PDB structures.
28 CELLULAR RESPIRATION

Table 2  Inputs and outputs from 12-carbon lactose to 2-carbon acetyl

groups and CO2.
inputs outputs
1 lactose (12 carbons) 4 CO2 + 4 acetyl groups (2 carbons each)
4 coenzyme A 4 acetyl-coenzyme A (acetyl groups from above)
4 ATP + 8 ADP 4 ADP + 8 ATP (net 4 ATP)
8 NAD+ 8 NADH

Source: Original art from public domain information.

values can be doubled for each 12-carbon lactose oxidized through gly-
colysis and the production of acetyl-CoA.
When looking at Figure 11, the linearity of this biochemical path-
way is striking. Substrate 1 leads to substrate 2, which leads to substrate
3 all the way down to the formation of pyruvate and later CO2 and
an acetyl group. Cellular respiration is regulated, and all species have a
variety of homeostatic mechanisms to regulate the speed of glycolysis.
How are the enzymes of glycolysis regulated to maintain energy homeo-
stasis? Consider this analogy. When working in a group on a common
task, the group’s overall rate of production is constrained by the rate of
the slowest step. More specifically, the rate of any given step is regu-
lated by one of two constraints: the rate of performing the task and the
availability of substrate. Let’s extend the analogy to clarify the critical
concept of regulating the rate of a large, multi-step process. Five people
want to package school supplies to send to Haiti where students lack
basic supplies. In each package, the Haitian students will find one pencil
sharpener, eight pencils, one handwritten note, two erasers, and a ream
of paper. Putting the pencil sharpener in is very fast, but it takes longer
to count out eight pencils, so the second step is slower than the first. In
our analogy, the supplies are similar to substrates, and the five people are
like enzymes. For the first two steps, enzyme 1 “sharpener-ase” is very
fast but enzyme 2 “pencil count-ase” takes longer to perform its task, so
the rate of the first two steps is limited by the rate of the second en-
zyme. The next step in the pathway is writing a note, which is the slowest
of the remaining steps, and therefore the entire assembly line is rate lim-
ited by the “write-ase” step. Once the note is written, putting the eras-
ers and paper into the package are very rapid. The speed of the last two
steps is limited by the availability of their substrates (erasers and paper).
 Converting Common Foods into Energy 29

substrate 1
e1
2 22 2 2
2 22
2 2 22

PFK
intermediates 2-7
3
e3
4
e4
5
e5 enzyme identification enzyme ( m M) substrate (m M)
e1 n.a s1 near 0
6
phosphofructokinase n.a s2 1,500
e6
e3 810 s3 80
7 e4 1,400 s4 80
7 7
7 e5 130 s5 50
PK e6 540 s6 20
8 = final product pyruvate kinase 170 s6a 65
A B

Figure 12  Homeostatic regulation of glycolysis. A, Overview of a

multi-enzyme pathway modeled on glycolysis plus the production of
CO2 and acetyl-CoA from Figure 11. B, Table with the concentration
of glycolysis enzymes and their substrates (s1–s7).
Source: Panel A, original art. Panel B modified from Srivastava and Bernhard, 1987; their table 1.

Biochemical pathways are very similar to a five person team working on

a common task. The rate of any step in a series of linked events is deter-
mined by the speed of the enzyme and the availability of substrates.
In Figure 12, it is evident that enzyme 2, phosphofructokinase
(PFK), is the slowest enzyme because its substrate has accumulated
behind it, just like the boxes of school supplies would be stacked up at
the note-writing station. Compounding this backlog of intermediate 2
is that enzyme 1 is the fastest enzyme in the entire chain as indicated
by its near zero substrate concentration in Figure 12B. Having almost
no glucose inside the cells indicates the 6-carbon sugar is immediately
phosphorylated once it reaches the cytoplasm of a cell (see Figure 11 for
details). The instantaneous investment of an ATP to produce a phos-
phorylated sugar ensures that the glucose stays inside of the cell because
 Free ebooks ==> [Link]

30 CELLULAR RESPIRATION

the phosphate group blocks its ability to leave the cell. All of the other
enzymes are substrate limited, as can be seen by the low concentrations of
their substrates in Figure 12B. As soon as substrates 3 through 6 are avail-
able, they are very quickly sent down the line to terminate at substrate 7,
pyruvate. Given that PFK is the slowest enzyme in the entire pathway,
PFK is the ideal enzyme to modulate in order to control the overall rate
of glycolysis. If the cell needs more ATP and NADH, then speed up
PFK. If the cell has sufficient levels of molecular potential energy, then
slow down PFK and let intermediate number 2 accumulate further. It is
worth noting that substrate 7, pyruvate, accumulates not because enzyme
8 is slow, but because pyruvate must be imported into the mitochon-
dria, which delays the conversion of pyruvate into CO2 and acetyl-CoA
(the final product 8).
PFK is the ideal enzyme to regulate glycolysis, but what molecules
can modulate PFK? In the early 1960s, biochemists discovered that PFK
activity is modulated by many molecules involved in cellular respiration.
Investigators measured the activity of PFK incubated with its substrate
alone (control) or its substrate plus citrate, a synonym for the high-
energy molecule citric acid. In another experiment, the biochemists
added citrate to PFK and then after 7 minutes, they added the low-energy
molecule AMP to the reaction. A different lab group tested the effects of
ATP on PFK’s activity by measuring the rate of product formation as a
function of ATP concentration.
ATP is consumed by PFK because PFK is a kinase. Kinases transfer
a phosphate from ATP and covalently attach the phosphate to produce a
new molecule, a phospho-sugar in this case. The need for ATP causes the
complex data for regulating glycolysis. PFK requires a small amount of
ATP to function, but high concentrations of ATP inhibit PFK. The ho-
meostatic regulation of PFK by ATP is evidence of a finely tuned enzyme
that is activated by low concentrations of ATP (0.25 mM) and inhibited
by high levels of ATP (1 mM). When a cell contains ample potential
energy in the form of ATP, PFK slows down and stops producing more
ATP. When the cellular supply of ATP gets as low as 0.25 mM, PFK activ­
ity increases until enough ATP is produced to shut down PFK activity
again. ATP is part of a negative feedback loop, which is also an emergent
property. PFK shows remarkable sensitivity to the cell’s potential energy

[Link]
 Converting Common Foods into Energy 31

status and is a prime example of molecular homeostasis. High levels of

AMP accelerated PFK’s activity but the cellular respiration intermediate
of citric acid inhibited PFK. When AMP and citrate are both added to
PFK, AMP overrides the inhibitory effects of citrate either by displacing
citrate or by binding to a different site on PFK and allosterically reducing
citrate’s inhibitory effect.
A group of Italian biologists documented a striking difference be-
tween rat PFK isolated from rapidly growing cancer cells and PFK
isolated from noncancerous cells. The relative activity of PFK in the pres-
ence of a wide range of citrate was compared to the level of PFK activity
in the absence of citrate. In rats and perhaps other mammals, PFK from
cancer cells does not respond to citrate the same way as PFK from non-
cancerous cells. PFK from cancer cells is less sensitive to citrate inhibi-
tion and continues to catalyze its biochemical reaction at half its normal
rate when PFK from noncancerous cells is only working at 10% of its
normal capacity. Maintaining a high rate of glycolysis ensures that the
rapidly dividing cancer cells will always have ample ATP, NADH, and
acetyl-CoA despite elevated levels of citric acid. It would be interesting to
sequence the cancer cell’s PFK gene to deduce from the codons whether
the enzyme protein has an altered amino acid sequence that blocks
citrate’s normal effect. Alternatively, the PFK promoter sequence could
be mutated in cancer cells and become over transcribed. A dominant
mutation to produce more PFK mRNA could permit PFK in cancer cells
to continue glycolysis despite elevated citric acid. Either way, hypothesiz-
ing a mutation in cancer cell PFK gene is logical and testable which is
what professional biologists do—design experiments.
Homeostasis of cellular respiration uses many molecular feedback
loops where the products of glycolysis affect the rate of glycolysis. PFK is
the rate limiting step of glycolysis due to its key role as the slowest enzyme
in glycolysis. Beta oxidation and amino acid deamination are also regu-
lated to maintain homeostasis of cellular respiration to convert food into
energy-rich molecules. Life is a struggle against the second law of thermo-
dynamics, which states everything in the universe tends toward disorder
unless energy is applied. To produce the energy required by cells to main-
tain order, protein enzymes oxidize fatty acids, proteins, and carbohy-
drates into 2-carbon bits plus the useful potential energy intermediates of
32 CELLULAR RESPIRATION

ATP, NADH, FADH2 and acetyl-CoA. ATP can be used to meet most of
the cell’s energy needs, but NADH, FADH2, and acetyl-CoA still contain
a lot of potential energy (see Table 1). ATP hydrolysis has a ΔG of only
−32 kJ/mole, whereas NADH has ΔG of −220 kJ/mole and glucose has
ΔG of −2873 kJ/mole. Because energy cannot be created or destroyed,
much of the potential energy from glucose is retained in the NADH,
FADH2 and acetyl-CoA molecules. Chapter 3 will address what happens
to the two carbons in acetyl-CoA, and Chapter 4 will address the majority
of ATP production.

Ethical, Legal, Social Implications:

Memorizing Details Obscures Learning
Diagrams similar to Figure 11 are probably familiar. Some students are
required to memorize the names of enzymes or structures of sugars in
glycolysis. When memorizing details, students (and some teachers) fail
to notice the mechanisms that regulate PFK or the homeostasis of ATP
in cells. Very few students know that beta oxidation and deamination
produce 2-carbon molecules just like the final step after glycolysis. No
student remembers all of the steps they had to memorize for glycolysis.
Psychologists have demonstrated repeatedly that when people perform
challenging, detail-oriented tasks, they fail to notice larger, significant
events. In a classic experiment, college students at the University of
Illinois were asked to watch a movie and count how many basketball
passes were made by only those students wearing white T-shirts. There
are six students in the movie, three with white T-shirts, and all of the
students are tossing two basketballs. After reporting their results, the stu-
dents were asked, “Did you notice anything unusual?” Only half of the
students noticed the person in a gorilla suit that walked right through the
middle of the students passing balls. It is easy to miss important events
(such as, energy homeostasis) when concentrating on a difficult task (such
as, memorizing the details of glycolysis).
In many studies on learning, investigators have repeatedly found that
the most effective means to learn information is a guided exploration,
which is the foundation of this book. Readers are not told what to learn,
but are guided to interpret experimental data in order construct into a
 Converting Common Foods into Energy 33

personal understanding. At various places in this book, commonly held

misconceptions have been presented about biology and then the reader
is asked to construct a correct understanding of the concept. Based on
years of research, we know that only when misconceptions are addressed
directly can a student release the misconception and embrace the correct
understanding.
In 2007, two educators in Turkey tested sixth grade life science stu-
dents on concepts related to the cell. One group of students was provided
with information in a traditional, passive format, and the other group of
students was guided through active learning exercises similar to what is
done in this book: Interpret information to construct a personal under-
standing. The two groups of students achieved the same scores immedi-
ately after the lesson was covered. However, 2 weeks later, those students
guided through active learning remembered significantly more (p < 0.05)
than those students who listened passively. Richard Mayer published a
retrospective study of 50 years worth of education literature. In his
2004 report, Mayer found that unguided exploration through hands-on
learning and class discussion did not enhance learning and often led
to frustration. Students benefited from guided exploration of data that
allowed them to construct their own knowledge and allowed them to
understand the concepts at a deeper level and for longer times.
In 2008, a pair of American psychologists conducted an experiment
to answer three questions:

1. Which strategy enhances learning and memory more: repeated

studying or frequent self-testing?
2. Can students accurately assess their own learning and predict success
on a graded test?
3. Does the rate of forgetting information correlate with the initial rate
of learning the information?

College students were randomly placed into one of four experimental

groups and asked to memorize a list of paired foreign words (Figure 13).
After an initial test, students were assigned to be in one of four groups
that only studied repeatedly, only practiced by verbal drilling and self-
testing, or a combination of these two activities, as indicated in Figure 13.
34 CELLULAR RESPIRATION

0.8

0.7

proportion recalled
0.6

0.5

0.4

0.3

0.2
study + – + –
testing + + – –
learning activities

Figure 13  Comparison of self-testing versus studying for long-term

recall. Students were given a list of paired foreign words to memorize.
Those who repeatedly self-tested were able to remember three times
as many words compared to those who studied but were not repeatedly
tested. 1 indicates the activity was performed; − indicates the
activity was not performed; error bars are standard error of the means.
Source: Modified from Karpicke and Roediger, 2008; their figure 2.

The data show that when students repeatedly tested themselves and were
forced to recall the information, they answered about 80% of the test
questions correctly. Students who merely studied repeatedly answered
only about 30% of the test questions correctly. Surprisingly, all four cat-
egories of students predicted that they would get only 20% of the test
questions. These data show that students are often bad at predicting out-
comes on tests they take. Furthermore, the initial learning rate of the
word pairs for all four experimental groups was identical and yet the for-
getting rate varied substantially as seen in the data.
What was learned from the movie of the gorilla among the students
throwing basketballs? Memorizing steps may be easier to accomplish in
the short run and easier to test, but memorization of detailed steps within
a bigger process tends to obscure big picture and important trends. Fur-
thermore, memorizing does not lead to long-lasting memory as well as
guided exploration does, as shown in Figure 13. In order to remember
information longer, such as energy homeostasis, practice self-quizzing or
use flash cards instead of simply rereading notes or a book. One last word
Free ebooks ==> [Link]

Converting Common Foods into Energy 35

about learning that may come as welcome news—sleeping is a necessary

step in order to construct long-term memory. Naps can be beneficial, and
studying all night will lead to rapid forgetting of information.

Bibliography
Colman R. Glutamate dehydrogenase (bovine liver). In Kuby SA. A study
of enzymes. Volume II. New York, 1991, CRC Press, pp. 174–192.
Fang J, Hsu BY, et al. Expression, purification and characterization of
human glutamate dehydrogenase (GDH) allosteric regulatory muta-
tions. Biochem J 363:81–87, 2002.
Frieden C. Glutamic dehydrogenase: the effects of various nucleotides
on the association-dissociation and kinetic properties. J Biol Chem
234(4):815–820, 1959.
Frieden C. Glutamate dehydrogenase: the regulation of enzyme structure
to the catalytic function. J Biol Chem 238(10):3286–3299, 1963.
Himoe A, Parks PC, Hess GP. Investigations of the chymotrypsin-
catalyzed hydrolysis of specific substrates. J Biol Chem 242(5):
919–929, 1967.
International Union of Pure and Applied Chemistry. Abbreviations and
symbols for chemical names of special interest in biological chemistry.
J Biol Chem 237(5):1381–1387, 1962.
Lehninger AL. The relationship of the adenosine polyphosphates to fatty
acid oxidation in homogenized liver preparations. J Biol Chem 157:
363–382, 1945.
Lowe G, Yuthavong Y. pH-dependence and structure-activity relation-
ships in the papain-catalysed hydrolysis of anilides. Biochem J 124:
117–122, 1971.
Malhotra OP, Prabhakar P, Sen Gupta T, et al. Phosphoglycerate-kinase-
glyceradehyde-3-phosphate-dehydrogenase interaction. Eur J Biochem
227:556–562, 1995.
McAndrew RP, Wang Y, Mohsen AW, et al. Structural basis for substrate
fatty acyl chain specificity. J Biol Chem 283(14):9435–9443, 2008.
Meldolesi MF, Macchia V, Laccetti P. Differences in phosphofructoki-
nase regulation in normal and tumor rat thyroid cells. J Biol Chem
251(20):6244–6251, 1976.

[Link]
36 CELLULAR RESPIRATION

Newsholme EA, Randle PJ. Regulation of glucose uptake by muscle.

Biochem J 93(3):641–651, 1964.
Northrop JH. Crystalline pepsin: isolation and test of purity. J Gen
Physiol 13(6):739–766, 1930.
Otto M, Heinrich R, Kuhn B, Jacobasch G. A mathematical model for
the influence of fructose 6-phosphate, ATP, potassium, ammonium
and magnesium on the phosphofructokinase from rat erythrocytes.
Eur J Biochem 49(1):169–178, 1974.
Parmeggiani A, Bowman RH. Regulation of phosphofructokinase activ-
ity by citrate in normal and diabetic muscle. Biochem Biophys Res
Commun 12(4):268–273, 1963.
Quick AJ. Quantitative studies of β-oxidation. J Biol Chem 77(2):
581–593, 1928.
Rösler Jens, Krekel F, Amrhein N, et al. Maize phenylalanine ammonia-
lyase has tyrosine ammonia-lyase activity. Plant Physiol 113(1):
175–179, 1997.
Strauss AW, Powell CK, Hale DE, et al. Molecular basis of human mito-
chondrial very-long-chain acyl-CoA dehydrogenase deficiency caus-
ing cardiomyopathy and sudden death in children. Proc Natl Acad Sci
92(23):10496–10500, 1995.
Sud M, Fahy E, Cotter D, et al. LMSD: LIPID MAPS structure database.
Nucleic Acids Res 35(Database issue):D527–D532, 2007.
Weber G, Convery HJ, Lea MA, et al. Feedback inhibition of key glyco-
lytic enzymes in liver: action of free fatty acids. Science 154(3754):
1357–1360, 1966.
Walker V, Taylor WH. Ovalbumin digestion by human pepsins 1, 3 and
5. Biochem J 176(2):429–432, 1978.

Ethical, Legal, Social Implications: Memorizing Details Obscures

Learning

Dogru M, Kalender S. Applying the subject “cell” through constructiv-

ist approach during science lessons and the teacher’s view. Journal of
Environmental & Science Education 2(1):3–13, 2007.
Donovan SM, Bransford JD, Pellegrino JW, editors: Committee on
Learning Research and Educational Practice. National Research
 Converting Common Foods into Energy 37

Council: How People Learn. Washington, DC, 1999, National

Academy Press.
Karpicke JD, Roediger HL. The critical importance of retrieval for learn-
ing. Science 319:966–968, 2008.
Kirschner PA, Sweller J, Clark RE. Why minimal guidance during in-
struction doe not work: an analysis of the failure of constructivist,
discovery, problem-based experimental, and inquiry-based teaching.
Educational Psychologist 41(2):75–86, 2006.
Mayer RE. Should there be a three-strikes rule against pure discovery
learning? The case for guided methods of instruction. American Psy-
chologist 59(1):14–19, 2004.
Simons DJ, Chabris CF. Gorillas in our midst: sustained in attentional
blindness for dynamic events. Perception 28:1059–1074, 1999.
 CHAPTER 3

Energy Extraction from

2-carbon Intermediates

Once proteins, lipids, and carbohydrates have been digested into 2-

carbon bits and modified into acetyl-CoA, most of the potential energy
has not yet been converted into ATP. Biochemists worked for years to
deduce how the 2-carbon molecules were metabolized further to extract
more energy in the form of ATP or NADH. While some were trying to
figure out how acetyl-CoA was digested to extract more energy, Hans
Krebs and colleagues wanted to determine how the 6-carbon molecule
citric acid, or citrate, was metabolized into a 4-carbon acid called oxa-
loacetic acid, or oxaloacetate (Figure 14). Acids carry a negative charge
and can be referred to as blank acid or blank-ate. Seven different acid
intermediates lie between citric acid and oxaloacetic acid. Along the
way, the multi-step pathway produces two CO2 molecules as waste and
many more NADH or FADH2 molecules. NADH oxidation has a ΔG of
−220 kJ/mole and FADH2 has ΔG of −201 kJ/mole. Therefore, the
production of oxaloacetic acid extracts a lot of potential energy in
the form of reduced electron carriers but only a single GTP. GTP carries
the same amount of potential energy as ATP and can be thought of as an
ATP equivalent.
Insights into cellular respiration led biologists to identify the enzymes
involved in all of the cellular respiration pathways, including the conver-
sion of citric acid into oxaloacetic acid as illustrated in Figure 14. Perhaps
it seems surprising to realize that the production of CO2 does not require
the consumption of any O2 molecules. The same was true when pyru-
vate was converted to acetyl-CoA and CO2 (see Figure 11). Along the
way, the enzyme reactions produced several electron carriers: 3 NADH;
1 FADH2, as well as 1 GTP. The GTP is only the second time ATP or its
40 CELLULAR RESPIRATION

citric acid = carbon atom

= oxygen atom

acid #2

acid #3

inputs outputs
+
+
acid #4 NAD NADH

acid #5 + NAD+ NADH

acid #6 GDP GTP

acid #7 FAD FADH2

acid #8

oxaloacetic acid NAD+ NADH

Figure 14  The 1936 understanding of molecular energy

conversion. Krebs began with this nine step, linear
understanding of how citric acid was metabolized into a
4-carbon oxaloacetic acid. Inputs and outputs appear to
the right of the acid produced at particular steps. Small
arrows point to covalent bond changes in each step; CO2
is circled. A large horizontal line marks the step blocked
by a poisonous metabolic inhibitor used by Krebs for his
research.
Source: Original art from common knowledge.
 Energy Extraction from 2-carbon Intermediates 41

400

= citric acid
micromoles in urine 300 = acid #4
= acid #6

200

acid #8 oxygen uptake oxygen uptake

100 added (calculated) (measured)
1 mole 6 moles 47.2 moles
2 moles 12 moles 69.4 moles
0
A acid #8 inhibitor oxaloacetic nothing B
injected injected acid injected
injected

Figure15  Metabolic products after injection of metabolic

intermediates. A, The indicated compounds were injected into rabbits
and their urinary output was measured (shaded bars). Metabolic acid
number and poisonous inhibitor are the same as in Figure 10.17. B,
Calculated and measured amount of oxygen consumed to convert acid
#8 to CO2 and H2O.
Source: Panel A modified from Krebs et al., 1937; figure 2. B. Panel B modified from Lehninger,
1970; table 17-1.

energy equivalent has been produced: glycolysis produced a net of two

ATPs per glucose. The conversion of a 6-carbon citric acid into a 4-carbon
oxaloacetic acid requires the removal of two carbons in the form of CO2.
Notice how the removal of CO2 is always coupled with the production
of an NADH.
Despite the delineation of this citric acid pathway, biologists were
still puzzled by some data that they could not explain (Figure 15). For
these experiments, investigators injected rabbits with various com-
pounds and then tested their urine to see where along the citric acid
pathway (Figure 14) the metabolites accumulated. As usual, start the
analysis by looking at the negative control data where nothing was in-
jected to see how much of each acid the rabbits normally urinate. Based
on the model in Figure 14, it would be predicted that injecting any acid
higher up the pathway would produce only acids further down the path-
way. If acid #2 is injected, it would be predicted that acids #3 through
#6 would accumulate in the rabbit’s urine, which is what the investiga-
tors found. However, the data in Figure 15A illustrate some puzzling in-
formation. Why would administration of acid #8 lead to accumulation
of acids #1 and #4? It makes sense that the inhibitor would lead to an
accumulation of acid #6, but acid #6 accumulated even when acids #7,
42 CELLULAR RESPIRATION

#8, and oxaloacetic acid were injected into the rabbits. Finally, when the
investigators gave the rabbits excess oxaloacetic acid, the last acid in the
pathway, citric acid accumulated. How? The data in Figure 15B added
to Kreb's confusion. Why would cellular respiration of acid #8 consume
more oxygen than was needed based on their calculations? Don’t feel
badly if confused by the data. Discovering the solution to these appar-
ent contradictions earned Hans Adolf Krebs a shared Nobel Prize.
Consider this analogy before trying to solve the puzzle that stumped
many biochemists from around the world. Picture someone in a long dark
hallway so that they can only see a couple feet in front or behind them.
The hallway represents the citric acid pathway that was poorly under-
stood until 1938 when Krebs and his collaborators published the solu-
tion. Standing in the dark hallway, the person throws a ball toward the
doorway in front of them, which represents oxaloacetic acid. No matter
where the person stands in the hallway to throw the ball, the ball always
comes to rest behind them. If they blocked the hallway at the sixth door,
the ball always stops at door six, even if they were standing at door 7 and
throw the ball toward door 9. When they stand at door number 8 and
throw the ball toward door 9, the ball stops behind them at door number
6 even though they are facing door 9. Remember, the hallway is poorly
lit, so the person cannot see the end of the hall just as Krebs and his con-
temporaries could not see the biochemical pathway from citric acid to
oxaloacetic acid.
Krebs was born in Germany where he earned his medical degree and
worked in hospitals treating patients. In 1933, Krebs left Germany for
Sheffield University in Britain because he had been fired from his previ-
ous job by Hitler's National Socialist government. Krebs redefined him-
self by becoming a bench scientist. Using the hallway analogy, perhaps
the reader made the same discovery Krebs did by realizing what seemed to
be a linear path was in fact a circular pathway (Figure 16). Krebs’ b­ rilliance
was not only his ability to conduct good biochemical experiments, but
more importantly he was able to see the same data as everyone else but in
a different light. He realized that like the hallway, the biochemical data in
Figure 15 indicated the pathway between citric acid and oxaloacetic acid
was not linear but circular. The loss of two carbons along the way cre-
ated an opportunity for the two carbons of acetyl-CoA (produced from
 Energy Extraction from 2-carbon Intermediates 43

CoA + citric acid

oxaloacetic
acid e e
1 NADH +

e + 1 NAD+ e
CoA

e = carbon atom
= oxygen atom 1 NAD+ e
1 FADH2 +
e 1 FAD +
+ 1 NADH
1 GDP +
+ 1 GTP e e
+
+ 1 NADH

Figure 16  Discovery of “citric acid cycle.” Krebs deduced the cyclic
nature of the pathway from the data in Figure 15. Small arrows show
changes in covalent bonds; the box is the addition of acetyl group;
inputs are inside the circle and outputs are outside the circle.
Source: Original art from common knowledge.

lipid, protein, and carbohydrate degradation) to bind to oxaloacetic acid

(four carbons) and reform citric acid (six carbons). The citric acid cycle,
as Krebs called it, produced a recursive loop that allowed the small inputs
of Figure 15B to stimulate more than five times the oxygen consumption
as predicted. Introducing acid #8 allowed the cell enzymes to consume
all their stored acetyl-CoA until it was depleted. It seems appropriate for
Krebs to share his 1953 Nobel Prize with the biologist who discovered
CoA and its role in cellular respiration, another German physician who
left his homeland for America.
The number of NADH, FADH2, and GTP produced can be
counted to verify the output of a citric acid cycle; it is the same number
as when people mistakenly thought the pathway was linear. It can also
be verified that no molecular O2 is consumed despite the fact that the
metabolic acids are being further oxidized as they move from citric acid
to oxaloacetic acid. By the end of the citric acid cycle, all the potential
energy of fatty acids, proteins, and carbohydrates has been harvested
into NADH, FADH2, and GTP or ATP with the waste carbons com-
pletely oxidized to CO2. No oxygen has been consumed in the form of
O2, and yet every organism requires oxygen to perform aerobic cellular
respiration. Like a good detective, Albert Lehninger was determined
to identify the location of oxygen consumption and the final stages of
44 CELLULAR RESPIRATION

Table 3  Subcellular fractions tested for the ability to carry out citric
acid cycle and consume oxygen.
source of protein substrate oxygen substrate product
protein added added consumed consumed formed
mitochondria 2.7 mg no 6.1 moles n.d. 0.9 moles
mitochondria 2.7 mg yes 23.2 moles 8.0 moles 8.5 moles
nucleus 3.5 mg no 1.5 moles n.d. 0.3 moles
nucleus 3.5 mg yes 2.3 moles 0.9 moles 1.1 moles
cytoplasm 4.5 mg no 1.3 moles n.d. 0.2 moles
cytoplasm 4.5 mg yes 1.0 moles n.d. 0.2 moles

Source: Modified from Kennedy and Lehninger, 1949.

Table 4  Homeostatic regulation of citric acid cycle.

step inhibitors activators
pyruvate acetyl-CoA + CO2 ATP, acetyl-CoA, NADH, fatty acids AMP, CoA, NAD+, Ca2+
acetyl-CoA + oxaloacetate citric acid NADH, citric acid, ATP ADP
acid #3 acid #4 ATP ADP, Ca2+
acid #4 acid #5 acid #5, NADH Ca2+
acid #8 oxaloacetate NADH acid #8

Source: Original art from common knowledge.

cellular respiration (Table 3). Lehninger subdivided rat liver cells into
three fractions: nucleus, cytoplasm, and mitochondria. To each fraction,
the biochemist added or withheld 2 mM fatty acid as a metabolic sub-
strate and then measured oxygen consumption, substrate consumption,
and product formation. From his results, it became the mitochondria
contained the enzymes responsible for the oxygen-consuming phase of
cellular respiration.
Table 4 summarizes the results from many experiments measuring en-
zyme reactions involved in the citric acid cycle. Converting food to energy is
essential for survival and therefore each pathway is carefully regulated by ho-
meostatic feedback loops. Biochemists often try to determine what metabo-
lites influence enzymatic reactions to either increase or decrease the rate of
the reaction. Muscle is often used to study cellular respiration in part because
it is easy to obtain large amounts from any given animal. Because of this
procedural bias toward muscle, investigators often test the effects of Ca2+
which is critical for the regulation of muscle contraction and the subsequent
consumption of ATP. Therefore, biochemists wanted to know whether
muscle contraction might be linked to homeostasis of cellular respiration.
Free ebooks ==> [Link]

Energy Extraction from 2-carbon Intermediates 45

Based on the data in Table 3, oxygen consumption takes place within

mitochondria because this fraction is the only source of enzymes that
consumed substrate and oxygen to generate a product. Lehninger knew
that beta oxidation of fatty acids into 2-carbon intermediates does not
consume oxygen, so his experiments were measuring the third and final
phase of cellular respiration covered in Chapter 4. Table 4 reveals that the
energy-rich products of cellular respiration (such as ATP, NADH, and cit-
ric acid) inhibit metabolic enzymes, whereas substrates that contain less
energy (such as ADP, CoA, and NAD+) activate the same enzymes. The
energy-rich metabolic intermediates function as part of negative feedback
loops when product accumulation inhibits more product formation.
Furthermore, high concentrations of low-energy substrates stimulate the
enzymes to catalyze their reactions faster.
Muscles require elevated cytoplasmic Ca2+ and vast amounts of ATP in
order to contract. Therefore it is understandable why it would be adaptive
for muscle cells to enhance cellular respiration when Ca2+ levels increase.
Perhaps it is less obvious why fatty acids block the conversion of pyruvate
to CO2 plus acetyl-CoA and NADH. Look back at Figure 7 to recall
that beta oxidation of fatty acids produces FADH2, NADH, and acetyl-
CoA for each pair of carbons removed. Therefore, if cells contain a lot of
fatty acids, they don’t need to consume pyruvate to produce one more
NADH when they will soon benefit from multiple copies of NADH
and FADH2 when acetyl-CoA brings two carbons into the citric acid
cycle.
Selective advantage during natural selection is determined by phe-
notypes that improve an individual’s ability to survive and reproduce.
Because nearly every cellular activity requires energy to maintain molecu-
lar order within cells, regulation of pyruvate production appears to be
adaptive in mammals and perhaps other organisms too. Look back at
Figure 10 and recall the allosteric regulation of PFK to see that prod-
ucts from the citric acid cycle also inhibit glycolysis and deamination of
amino acids. The conversion of the 2-carbon donor acetyl-CoA into CO2,
GTP, and reduced electron carriers is a critical step in the molecular ex-
traction of potential energy from food. Homeostatic regulation of energy
flow within a cell has been the subject a billion of years of natural selec-
tion because every mitochondrion on the planet is involved in cellular

[Link]
46 CELLULAR RESPIRATION

respiration, including oxygen consumption. Chapter 4 will explain where

and how enzymes consume oxygen and produce ATP.

Bibliography
Krebs HA, Salvin E, Johnson WA. The formation of citric acid and
a-ketoglutaric acids in the mammalian body. Biochem J 32(1):
113–117, 1938.
Krebs HA. The role of fumarate in the respiration of Bacterium coli com-
mune. Biochem J 31(11):2095–2124, 1937.
Kennedy EP, Lehninger AL. Oxidation of fatty acids and tricarboxylic
acid cycle intermediates by isolated rat liver mitochondria. J Biol
Chem 179(2):957–972, 1949.
Morales MF. Some thermo analytic studies of organ and whole animal
respiration. J Gen Physiol 26(4):381–389, 1943.
Free ebooks ==> [Link]

CHAPTER 4

ATP Production from

Digested Foods

The citric acid cycle, which takes place inside the mitochondria, con-
verts energy from the 2-carbon acetyl-CoA intermediate into GTP and
the electron carriers, or reducing agents, NADH and FADH2. Glycolysis
happens in the cytoplasm but the metabolism of pyruvate into acetyl-
CoA and NADH takes place in the mitochondria and does not consume
oxygen. Lehninger demonstrated that oxygen consumption and the final
phase of cellular respiration takes place inside the mitochondria. There-
fore, cytoplasmic NADH must be imported into the mitochondria, which
requires some energy. However, once inside the mitochondria, eukaryotic
cells have concentrated a lot of reducing power in the form of NADH
and FADH2. The last remaining question biologists needed to answer to
understand the fundamentals of cellular respiration was how the potential
energy in the reducing agents (NADH and FADH2) is converted into
new covalent bonds connecting ADP with inorganic phosphate, Pi.
One of the keys to understanding how mitochondria convert the
potential energy in NADH and FADH2 into ATP was realizing the
pH inside mitochondria differed from the pH of the cytoplasm. Using
modern methods, investigators have confirmed the historic lessons by
engineering fluorescent proteins that alter their color depending on the
surrounding pH. These pH indicator proteins are located in the lumen
of the mitochondria, also called the mitochondrial matrix. The change
in colors is not readily visible to our eyes, so computer programs quantify
the pH-induced color shift. To verify that the pH of the mitochondrial
matrix is different from the cytoplasm, investigators monitored the pH
inside mitochondria and then added a drug to insert H+ ion channels in
the inner mitochondrial membrane. The added ion channels would let

[Link]
48 CELLULAR RESPIRATION

H+ ions flow down their concentration gradient until the cytoplasm and
the matrix have the same pH of 7.0. Cell biologists had demonstrated
previously that the outer mitochondrial membrane is rather porous
and does not exclude ions from the intermembrane space between
the two membranes of mitochondria. Based on experimental evidence,
the biologists confirmed the pH of cytoplasm was about 7, which is the
same pH of the intermembrane space because the outer mitochondrial
membrane is porous to ions. The mitochondrial matrix in this experi-
ment had a pH slightly higher than 8, which meant the cytoplasm had
a H+ ion concentration that was about ten times higher than the mito-
chondrial matrix. Remember that the pH scale is a log10 scale so every
one pH unit of change is a tenfold change in H+ concentration.
The next experiment was the critical one to connect reducing power
of NADH and FADH2 to the pH inside mitochondria. Biochemists
produced membrane vesicles devoid of any mitochondrial proteins and
then experimentally inserted a protein complex from mitochondria hy-
pothesized to play a role in ATP production. The mitochondrial protein
complex was added to the experimental vesicles so that the luminal side
of the vesicle was functionally equivalent to the mitochondrial matrix
and outside the vesicles was functionally equivalent to the mitochondrial
intermembrane space. The investigators measured the pH of three solu-
tions containing equivalent preparations of these experimental vesicles
before adding one of three treatments: NADH alone; NADH plus H+
ion channel; NADH plus a drug that inhibits the mitochondrial protein’s
function. The experimental vesicles containing the mitochondrial protein
complex transported H+ ions from the lumen to outside the vesicles when
NADH was added. Inside cells, the mitochondrial matrix has a lower H+
ion concentration (higher pH) than the cytoplasm because NADH pro-
vided the power to export H+ ions. The movement of H+ ions requires
a reducing agent, and the rate of ion movement is substantially reduced
when the mitochondrial protein is inhibited by the added drug. The pH
of the vesicle lumen was unnaturally lower than the pH seen in living
cells due to the non-natural conditions of the experimental vesicles. The
pH of the buffered solution outside the vesicle was near 6.5 because the
H+ ion channel added to the vesicles allowed the ions to become equally
distributed on both sides of the vesicle membrane. The data indicate that
 ATP Production from Digested Foods 49

when given a reducing agent, mitochondria transport H+ ions out of the

matrix and into the cytoplasm. Removing H+ ions raises the mitochon-
drial matrix pH to about 8 compared to cytoplasmic pH of about 7.
After many more years of incremental experimentation, biochemists
were able to compile their understanding into a comprehensive model of
the electron transport chain embedded in the inner mitochondrial mem-
brane (Figure 17A). A series of four, multi-subunit protein complexes
work as an integrated unit to oxidize NADH (into NAD+) and FADH2

4 H+ 4 H+ 2 H+
outside mitochondria
2 e– 2 e– 2 e–
I II III IV

inside mitochondria
NADH NAD+ FAD
1
O2 H2O (matrix)
2
FADH2 2H
A

= 0 ppm CO
= 50 ppm CO
= 100 ppm CO
180 = 500 ppm CO
160
140
percent activity

120
100
cytochrome c 80
60
40
20
0
I II III IV*
B hemoglobin C electron transfer complex

Figure 17  Electron transfer pathway in mitochondrial membrane.

A, Four protein complexes contain hemes and transfer electrons from
NADH and FADH2 donors to the final acceptor molecule. B, Atomic
structures of two similar hemes from cytochrome c of complex IV and
human hemoglobin. C, Effect of carbon monoxide (CO) on electron
transfer by the four complexes shown in A, +/− standard deviation.
* indicates p < 0.001.
Source: Panels A & B from common knowledge–original art. Panel C modified from Alonso
et al., 2003; part of their figure 2. Alonso, Jose-Ramon, Francesc Cardellach, et al. 2003.
Pharmacology and Toxicology. Vol. 93: 142–146. Carbon Monoxide Specifically Inhibits
Cytochrome C Oxidase of Human Mitochondrial Respiratory Chain.
50 CELLULAR RESPIRATION

(into FAD) and transport the electrons down the chain from complex I
to complex IV. Complex I is more electronegative than NADH, which
causes complex I to become reduced by NADH that becomes NAD+. In
the process of accepting the pair of electrons from NADH and passing
them onto complex II, complex I transports 4 H+ ions from the matrix
to the intermembrane space of the mitochondria. The electrons continue
their journey toward more and more electronegative complexes and pro-
vide the energy necessary to pump H+ ions up their concentration gradi-
ent into the cytoplasm where H+ are accumulating. However, FADH2
cannot reduce complex I, so its electrons begin their movement through
the electron transport chain starting at complex II. The movement of
electrons from FADH2 is independent of the electrons from NADH, and
vice versa.
A critical function of the proteins within the electron transport chain
is the ability to temporarily bind electrons and then pass them on. Amino
acids are not well suited for this task, but a familiar structure is—heme
(Figure 17B). Cytochromes are a family of proteins that are part of
the electron transport chain. These cytochromes contain heme molecules
the same way that hemoglobin protein in red blood cells carries heme
groups. In fact, all the hemes are nearly identical in structure with the
only differences being well away from the four nitrogen atoms that
coordinate the central iron ion. Heme can bind to oxygen and electrons
because of the positively charged iron ion at the center of the heme.
Unfortunately, heme can also bind to carbon monoxide (CO). CO
can bind to the heme in hemoglobin, but can CO bind to the heme in
cytochrome proteins and affect electron transport? To determine whether
CO could bind to the hemes of cytochromes within the electron trans-
port chain, biochemists tested the ability of CO to inhibit each of the
four protein complexes (Figure 17C). They tested the ability of each com-
plex to transport electrons in the presence of increasing concentrations of
dissolved CO, from 0 parts per million (ppm) to 500 ppm. From the data
in Figure 17C, one can see that only complex IV is inhibited by CO, as
indicated by the significance of p < 0.001 on the graph. When CO binds
to the heme in complex IV, electrons cannot move from complex III and
the entire chain becomes clogged. It is not a coincidence that the only
complex inhibited by CO is also the only complex where O2 binds to a
 ATP Production from Digested Foods 51

heme. CO binds to heme with higher affinity than O2 binds, which ex-
plains why CO poisoning is fatal. CO could kills by blocking the electron
transport chain which shuts down the entire electron transport chain.
Interestingly, cigarette smoke contains CO, which may contribute to its
toxic effects on lung cells.
Complexes I, III, and IV pump H+ ions while moving electrons
down the pathway toward oxygen. Complexes I and III transport 4 H+
ions each, but complex IV only transports 2 H+ ions. NADH reduces
complex I and therefore each pair of electrons from NADH causes 10
H+ ions to move out of the matrix. FADH2 skips complex I and reduces
complex II, and so the two electrons from FADH2 only provide enough
energy to move 6 H+ ions. The number of H+ ions pumped by electrons
from NADH and FADH2 differ because the reducing agents deliver their
electrons to different complexes within the chain.
The amino acid composition of the proteins in the four complexes
determines whether dissolved gases have access to the hemes. Under nor-
mal conditions, oxygen binds to the heme when a pair of electrons ar-
rives. The electrons bind to the oxygen, which would produce an atom
with a charge of negative 2, but two H+ ions from the mitochondrial
matrix bind to the oxygen ion to form water, H2O. Each atom of O2 is
reduced by a pair of electrons and so that one oxygen molecule produces
two water molecules (2H2O), which consumes four H+ ions. The use
of H+ ions from the mitochondrial matrix to form water explains why
complex IV transports only half as many H+ ions as complexes I and III.
The reducing power of NADH and FADH2 cause the mitochondria to
establish a H+ ion gradient with more H+ ions outside the mitochondria.
Animals need oxygen because it is consumed during cellular respiration as
the final electron acceptor and produces water as a waste product.
After breaking all digested food into 2-carbon bits, running around
the citric acid cycle and producing water from O2 and hydrogen ions,
cells have only produced a net of 2 ATP and 2 GTP for every glucose con-
sumed. The only other potential energy left is the H+ ion gradient across
the inner membrane of the mitochondria. From this starting point in
1961, Peter Mitchell at the University of Edinburgh in Scotland proposed
the chemiosmotic hypothesis to explain how the H+ ion gradient is used
to produce ATP. His hypothesis earned him a Nobel Prize in Chemistry
52 CELLULAR RESPIRATION

in 1978 when it became clear that the H+ ion gradient drives ATP syn-
thesis. Now Mitchell’s proposal is called the chemiosmotic theory because
it has been validated to the point that no one doubts its validity.
The key to understanding the chemiosmosis is understanding the
structure of the ATP synthase, a multi-subunit protein embedded in the
inner mitochondrial membrane (Figure 18A). Chapter 1 reviewed ΔG,
which quantifies the potential energy to do work. Recall that steam can
be used to push turbine blades to produce electricity. This same general
design evolved very early in the evolution of life because archaea, bacteria,
and eukaryotes all share the same general mechanism for the production
of ATP. Like steam, the H+ ion gradient spins a central protein shaft
to convert ADP plus inorganic phosphate (PO42 or Pi) into ATP. The
H+ ions rush down their concentration gradient into the matrix, with its
higher pH, via a small channel within the base of the ATP synthase. The

H+ ions

3
outside
mitochondria
1
2

inside
mitochondria ADP
(matrix) + Pi ATP
ATP ADP
+ Pi H+ ions

A B

Figure 18  ATP synthase spins like a turbine to produce ATP. A,

Molecular structure of ATP synthase is very similar to a human-made
turbine. B, Immobilized ATP synthase spins its base backward when
consuming ATP as visualized by attached actin rod.
Source: Panel A from common knowledge and PDB file. Panels B modified from Sambongi et al.,
1999; figures 1 and 2a. Sambongi, Yoshihiro, Yuko Iko, et al. 1999. Mechanical rotation of the
c subunit oligomer in ATP synthase (F0F1): Direct observation. Vol. 286: 1722–1724. Reprinted
with permission from AAAS.
 ATP Production from Digested Foods 53

rotating portion is the shaft that holds the “bumpy doughnut” slightly
away from the membrane. For many years, biochemists had used indirect
means to measure the rotation of the ATP synthase that protrudes into
the matrix, but in 1998, a group of Japanese biophysicists designed an
experiment that allowed them to film the rotary motion. For technical
reasons, rather than attaching the base of the ATP synthase to a micro-
scope slide, they attached the bumpy doughnut complex upside down
to the glass slide (Figure 18B). Because the base is uniform in shape
and very small, the Japanese biophysicists attached different lengths of
fluorescent actin rods to the ATP synthase base so that they could watch
the actin rods spin around. Years ago, biochemists discovered that in the
absence of an H+ ion gradient, ATP synthase could consume ATP and
spin its turbine-like base backward. The actin only rotated (backwards)
when the investigators supplied the modified ATP synthase with ATP
for fuel. In the presence of ATP, the biophysicists made movies of what
they saw through a very powerful light microscope. As they watched,
they noticed that the actin rod sometimes paused at particular places
and found that the 360 degrees were partitioned into three equal parts.
To be certain what they were watching was in fact a rotation of the ATP
synthase, they produced actin rods of different lengths and measured
the rate of rotation.
Sometimes it is hard to remember that even tiny molecules such
as ATP synthases are 3D objects that adhere to the physical laws of
nature, such as friction and centripetal force. If a person rotates in a
chair, extending their arms slows their rotation rate and bringing their
arms closer to the body speeds up their rotation. Likewise, a long actin
rod produced more resistance through the liquid medium than a short
rod attached to the base of the ATP synthase. Therefore, the ATP syn-
thase rotated faster with short actin rods and would rotate even faster
with no actin attached to the base of the synthase. Mitochondria in cells
rotate the shaft about 150 revolutions every second. The actin rod ex-
periment was designed to reveal the rotary motion of the ATP synthase
and used only part of the full protein complex. The portion that forms
the H+ ion channel was omitted from the experiment, because it does
not rotate. only the central shaft rotates but the “bumpy doughnut”
protein complex does not rotate.
54 CELLULAR RESPIRATION

H+ ions move from the cytoplasm into the mitochondrial matrix

when they pass through a small channel formed by several subunits of
the ATP synthase. The ions move down their concentration gradient by
random motion of diffusion from the area of higher H+ ion concentra-
tion and thus lower pH. After analyzing the movies and where the ATP
synthase paused, it appears that the ATP synthase shaft rotates in units of
120 degrees to produce one ATP.

Overview of Energy Homeostasis

Proteins, lipids and sugars are converted into 2-carbon molecules that
join the citric acid cycle. Electrons are delivered to the electron transport
chain and reduce O2 to water. The resulting H+ ion gradient is used
to drive the production of ATP. Energy production from raw materials
and energy consumption must be carefully controlled. Cells rely upon
allosteric interactions and feedback loops to maintain homeostasis of
cellular respiration (Figure 19). Arrows indicate when a compound ac-
tivates or accelerates the indicated step in cellular respiration. Blocking
symbols indicate which compounds inhibit or stop the indicated step of
converting food into ATP. The two toxins of cyanide and CO are added
to the figure to help understand why they are lethal regardless of what
type of food was consumed to produce ATP. And remember that GTP,
produced during the citric acid cycle (see Figure 16), will inhibit GDH
which means deamination is curtailed if other energy sources are plen-
tiful (see Figure 10). Furthermore, reduced deamination produces less
ammonia waste which protects the kidneys from unhealthy stress. Since
kidneys help regulate blood pressure, GTP production during the citric
acid cycle is a key player in overall health.
From Figure 19, it is clear that products of cellular respiration with
large amounts of potential energy to do work (such as, NADH, citric
acid, acetyl-CoA, and ATP) all inhibit or slow down cellular respiration.
Products that inhibit their own production contribute to negative feed-
back loops. Substrates for cellular respiration accelerate their own con-
sumption, but one cannot generalize that all the enzymes are allosterically
modulated by their substrates. The enhancement of substrate consump-
tion might be due to the increase availability of a rate-limiting substrate
Free ebooks ==> [Link]

ATP Production from Digested Foods 55

citric
acid AMP
amino acid O ATP
O

OH OH
R palmitic acid
lactose

GTP ATP
NADH acetyl-CoA
ADP
CoA

fatty acids Ca2+

acetyl-CoA acetyl-CoA AMP
ATP CoA
NADH NAD+

2C + = 6C

ATP Ca2+
NADH + CO2 ADP
citric
CO2 +
acid
paired electrons
(potential energy)

ADP
cyanide
ATP O2 = H2O
and CO

H+ ion gradient
(potential energy)

ADP
ATP
+ ADP = ATP
+ Pi

Figure 19  Homeostasis of cellular respiration. Products and

substrates regulate their own production by negative feedback loops
and allosteric modulation to increase the rate of substrate-limited
reactions. Arrows indicate enzyme activation and blocking symbols
indicated inhibitory signals.
Source: Original art based on common knowledge.

of a reaction. Nevertheless, when the ratio of substrates to products in-

creases, metabolism increases and molecular homeostasis is maintained by
producing more ATP. Of all the inhibitory products, ATP seems to have
a nearly universal inhibitory effect, which makes sense given the produc-
tion of ATP is the major end result of cellular respiration.

[Link]
56 CELLULAR RESPIRATION

Table 5  Calculations to determine efficiency of cellular respiration.

actual
ΔG number Σ ATP
energy possible ATPs ΔG respiration
source (kJ/mole) produced (kJ/mole) efficiency
ATP −32 1 −32 n.a.

FADH2 −201 1.5* −48 24%

NADH −220 2.5* −80 36%

glycine −1,008 12 −384 38%

lactose −5,746 64 −2,048 36%

palmiticacid −9,789 108 −3,456 35%

*partial ATPs account for energy lost due to importing energy source into mitochondria.

With a complete overview of cellular respiration, it would be interest-

ing to know how efficient the entire process is (Table 5). In the absence
of CO or cyanide, cellular respiration is about 35% efficient, which is
almost double the efficiency of a high-efficiency car (~20%). The mo-
lecular mechanisms used to convert lipid, proteins, and carbohydrates are
impressively efficient. With exposure to CO and cyanide, cellular respira-
tion would stop at the electron transport chain because both poisons bind
to complex IV. Once all the NAD+ and FAD were converted to NADH
and FADH2, the citric acid cycle would stop functioning, and cells would
not be able to make GTP either. Because glycolysis also requires NAD+ as
a substrate, it too would stop producing ATP. Therefore, CO and cyanide
would reduce the efficiency of cellular respiration to zero, and all cells
would die from a lack of ATP production.
Biological systems require energy to maintain structure and order
within cells. Multicellular organisms require additional energy to produce
and organize tissues and other levels of structure, which are required for
proper function. Cellular respiration is a molecular level example of ho-
meostasis that provides energy for all biological work. Cellular respiration
uses feedback mechanisms to regulate and maintain optimal efficiency
of converting food into the potential energy found in ATP. Time-depen-
dent processes regulate cellular respiration as shown by ATP inhibition
of energy production and the enhancement of cellular respiration by low
energy molecules, such as ADP and AMP. ATP is produced at the last
 ATP Production from Digested Foods 57

phase of cellular respiration and has the greatest inhibitory effect on all
the pathways. The size and environment within the mitochondria influ-
ences how the proteins function to produce ATP. Enzymes use proximity
in quaternary structures, such as the electron transport chain to facilitate
the movement of electrons. The ATP synthase shaft rotates very quickly in
the middle of the larger bumpy head where ATP is produced. Perhaps the
three pauses in ATP synthase per one rotation reflect the rate of substrates
diffusing to their binding sites. In short, molecular homeostasis of energy is
centered on the consumption of carbon sources as a means to convert the
covalent bonds of food into covalent bonds of ATP.

Ethical, Legal, Social Implications:

The Importance of Eating Vegetables
Having completed a study of the molecular homeostasis of energy, one
could be excused for reaching a mistaken conclusion—all a person needs
to eat is protein, fat, and sugar. These three food sources will provide a lot
of ATP, but every cell on the planet needs more than just carbon sources
for ATP production. Every cell requires what are called trace elements and
micronutrients. These components of a healthy diet include vitamins
such as A, B, C, D, and E, as well as the elements iodine, magnesium,
manganese, copper, zinc, iron, and selenium among many others. It is
impossible to obtain all of these nutrients from meat and potatoes, so
a healthy diet must include plant foods, such as fruits and vegetables.
Ironically, people also should eat a type of carbohydrate from which they
cannot extract any energy—cellulose, which most people call roughage.
Indigestible roughage is helpful in preventing colon cancer, because the
undigested cellulose prevents the build up of damaging compounds that
would remain in the colon otherwise.
On March 22, 1990, President Bush Sr. declared, “I do not like broc-
coli. And I haven’t liked it since I was a little kid and my mother made
me eat it. And I’m President of the United States and I’m not going to eat
any more broccoli.” Bush’s quote is perhaps the most famous example of
people who don’t like to eat vegetables, but the President quickly clarified
that he eats many other vegetables instead. Fruits and vegetables provide
nutrients that animals need to remain healthy (Table 6). Many Americans
 Free ebooks ==> [Link]

58 CELLULAR RESPIRATION

Table 6  Nutrients and the symptoms from deficiencies.

Source: Original art from common knowledge.

Table 7  Malnutrition causes and rates in

America.
micronutrient Americans deficient
vitamin E 93%
magnesium 56%
vitamin A 44%
vitamin C 31%
vitamin B6 14%
zinc 12%
folate  8%

Source: Open access data.

falsely believe malnutrition is not a problem in their country, but if one

considers nutrients and not calories, then many Americans do experience
malnutrition through deficiencies of trace elements and vitamins (Table 7).
In addition to requiring many nutrients, a body can suffer from too
much of a good thing. If a person eats a high fat diet and does not exer-
cise, that person will become obese. Eating too much protein can harm
kidneys, which can result in high blood pressure. If a person’s primary
protein intake is meat, then that person risks of too much cholesterol con-
sumption, and this can lead to heart disease. Overeating carbohydrates
leads to elevated sugar in the blood, which can cause arteries to become
less elastic and can increase the chances of a heart attack.
Energy is not the only molecular homeostasis occurring in cells. All
of the vitamins, minerals, and elements that a person needs to be healthy
are regulated as well. It may seem hard to cure all these diseases, but the
solution is quite simple. Adjust one’s diet according to [Link], and
everyone will be fine: Eat a balance of grains, vegetables, fruits, and dairy,
plus meats and beans.

[Link]
 ATP Production from Digested Foods 59

Bibliography
Abad MF, Di Benedetto G, Magalhaes PJ, et al. Mitochondrial pH moni-
tored by a new engineered green fluorescent protein mutant. J Biol
Chem 279(12):11521–11529, 2004.
Alonso JR, Cardellach F, Lopez S, et al. Carbon monoxide specifically
inhibits cytochrome c oxidase of human mitochondrial respiratory
chain. Pharmacol Toxicol 93(3):142–146, 2003.
Leung KH, Hinkle PC. Reconstitution of ion transport and respiratory
control in vesicles formed from reduced coenzyme Q-cytochrome c
reductase and phospholipids. J Biol Chem 250(21):8467–8471, 1975.
Llopis J, McCaffery JM, Miyawaki A, et al. Measurement of cytosolic,
mitochondrial, and Golgi pH in single living cells with green fluores-
cent proteins. Proc Natl Acad Sci 95(12):6803–6808, 1998.
Mitchell P. Coupling of phosphorylation to electron and hydrogen trans-
fer by a chemi-osmotic type mechanism. Nature 191:144–148, 1961.
Rees DM, Leslie AG, Walker JE. The structure of the membrane extrinsic
region of bovine ATP synthase. Proc Natl Acad Sci 106(51):21597–
21601, 2009.
Sambongi Y, Iko Y, Tanabe M, et al. Mechanical rotation of the c sub-
unit oligomer in ATP synthase (F0F1): direct observation. Science
286:1722–1724, 1999.
Stock D, Leslie AG, Walker JE. Molecular architecture of the rotary
motor in ATP synthase. Science 286:1700–1705, 1999.
Yasuda R, Noji H, Kinosita K Jr., et al. F1-ATPase is a highly efficient
molecular motor that rotates with discrete 1208 steps. Cell 93(7):
1117–1124, 1998.

Ethical, Legal, Social Implications: The Importance of Eating

Vegetables

Dowd M. ‘I’m President,’ So No More Broccoli! The New York Times (web
archive): [Link]
[Link]?scp=1&sq=bush%20broccoli&st=cse.
Accessed March 16, 2010.
Martin WF, Armstrong LE, Rodriguez NR. Dietary protein intake and
renal function. Nutr Metab 2:25–33, 2005.
60 CELLULAR RESPIRATION

Moshfegh A, Goldman J, Cleveland L. What we eat in America, NHANES

2001–2002: usual nutrient intakes from food compared to dietary
reference intakes. United States Department of Agriculture (USDA),
Agricultural Research Service (website): [Link]
research/publications/[Link]?SEQ_NO_115=184176.
Accessed July 6, 2014.
Tel Aviv University. How high carbohydrate foods can raise risk for heart
problems. ScienceDaily (website): [Link]
releases/2009/06/[Link]. Accessed March 16, 2010.
 Conclusion
Life requires energy to maintain structure and function. The second law
of thermodynamics states that all systems tend toward randomness un-
less energy is applied to maintain order. The first law of thermodynamics
states that energy cannot be created or destroyed only converted from one
form to another. Complex molecules containing many covalent bonds
are ideal sources of biological energy that can be used by cells to perform
work. When covalent bonds are broken, some of the energy is converted
to new covalent bonds within the reducing agents NADH and FADH2
that carry electron pairs to the electron transport chain. Energy is also
contained in ATP and GTP, which can be produced without the use of
the ATP synthase. Initially, the large molecules are broken down into
2-carbon molecules that flow into the citric acid cycle. By the end of the
citric acid cycle, all of the carbons are fully oxidized into CO2. Most of
the potential energy is stored in reducing agents. The electrons flow down
the electron transport chain to the final acceptor of O2, which produces
water by combining the electrons with H+ ions. As the electrons are shut-
tled toward oxygen, they provide the energy needed to pump H+ ions out
of the mitochondrial matrix and establish an electrochemical gradient.
The H+ ion gradient provides the power to rotate the ATP synthase to
produce ATP.
Each step along the pathway is regulated to maintain an appropriate
balance of substrates and products, which is the hallmark of homeostasis.
Cells maintain molecular homeostasis of energy flow by negative feed-
back loops and allosteric modulation of enzymes. Some reactions are rate
limited by the availability of substrate, which provides another mecha-
nism of maintaining homeostasis of energy flow. ATP is the most potent
inhibitor of cellular respiration, which works well because ATP is the
final product that determines the energy needs of a cell. Other meta-
bolic products also slow down the rate of cellular respiration, whereas
substrates can stimulate enzymes or accelerate reaction rates by increasing
the availability of limiting reagents. The timing of metabolite production
62 CELLULAR RESPIRATION

determines which steps of cellular respiration are stimulated or inhibited.

Enzymatic reactions that occur early in cellular respiration are regulated
by many molecules, whereas ATP synthase is only modulated by ATP and
ADP. The small size of the mitochondria and the ATP synthase dictate
how they function inside cells. Rotation of the ATP synthase is affected
by friction between the moving protein and the mitochondrial matrix.
The small volume of the mitochondrial matrix permits the quick estab-
lishment of H+ ion gradients from reducing agents. By passive diffusion,
H+ ions race through the ATP synthase, which uses a rotary motion to
produce new covalent bonds between ADP and Pi. The protein molecules
of cellular respiration regulate the energy flow from food to ATP.
 Glossary
acetyl-CoA. one product of cellular respiration where two carbons, an acetyl
group, are covalently attached to CoA.
allosterically modulated. a protein’s function is changed when any molecule or
element binds non-covalently to a protein and alters the protein’s shape.
ATP synthase. the multi-protein, mitochondrial complex that converts the
potential energy of an H+ ion gradient into ATP.
beta oxidation. the digestion of fatty acids into 2-carbon molecules, starting at
the acid side.
casein. most abundant protein in human milk.
cellular respiration. it describes the interwoven molecular processes used by cells
to convert food into energy that can perform biological work.
chemiosmosis. the H+ ion gradient across the inner mitochondrial membrane
drives the synthesis of ATP in the mitochondrial matrix.
citrate. six carbon molecule that starts the citric acid cycle for the extraction of
energy.
coenzyme A (CoA). a coenzyme and is consumed during enzymatic beta oxida-
tion of fatty acids.
coenzymes. organic molecules that are required for enzymatic reactions; they are
often covalently modified as part of the reaction.
cofactors. small, inorganic molecules, atoms, or ions that are required for enzy-
matic reactions.
deamination. the enzymatic removal an amine group (NH2) from an amino acid.
entropy. a measure of the degree of randomness or disorder of matter.
enzymes. enzymes perform many of the cell’s activities and can do so repeatedly
since they are not consumed as they function.
exons. portions of eukaryotic mRNA that are retained during mRNA splicing.
first law of thermodynamics. states that energy cannot be destroyed or created
but it can change forms.
free energy. in a biological context, it is the energy available to do work.
homeostasis. it maintain internal conditions within a range of acceptable extremes.
homozygous. homozygous individuals have two copies of the same version of a
gene.
introns. portions of eukaryotic RNA that are discarded during mRNA splicing.
kinase. enzymes catalyze the reaction of attaching a phosphate to covalently mod-
ulate its shape and function.
64 GLOSSARY

lactase. enzyme that begins the process of breaking down the sugar lactose for the
extraction of energy.
lactose. a sugar found in milk and composed of two simpler sugars.
limiting factor. determines the extent to which a process can proceed and prevent
excessive actions.
micronutrient. vitamins and minerals necessary for good health and not pro-
duced by the body; must be consumed in diet.
mitochondrial matrix. the lumen of the mitochondria that is inside both
membranes
mRNA. messenger RNA that was transcribed from a gene and is translated by the
ribosome into a protein.
negative feedback loop. it occurs when the product of a process results in a reduc-
tion of the same product being produced.
oxidized. the loss of electrons from broken covalent bonds
palmitic acid. most abundant fatty acid in human milk.
pedigree. diagram of families showing the relationships between male (squares)
and female (circles) members. Shapes that are shaded in exhibit a disease state.
peptide bonds. the covalent bond connecting two amino acids in a protein.
phosphofructokinase (PFK). slowest enzyme in the glycolysis metabolic path-
way; enzyme is regulated by allosteric modulators.
redox. reactions involve reduction and oxidation where one molecule is reduced
and a different molecule is oxidized.
reduced. the acquisition of electrons stored as covalent bonds
second law of thermodynamics. states that the universe has increasing disorder,
or entropy.
thermodynamics. the study of energy and mechanical power, including molecu-
lar level power.
work. the change of position or state of matter, such as moving an object or con-
verting a liquid to a gas.
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Index
Acetyl-CoA, 16, 20, 26 Digested foods, ATP production
Allosterically modulated, 24–25 from, 47–58
Amino acids, deamination of, 23–25 energy homeostasis, 54–57
ATP ethical, legal, social implications
conversion to ADP, 6–7 of, 57–58
production, from digested foods,
47–58 Eating vegetables, importance of,
synthase, 52–54 57–58
Electricity, 1, 4
Benzoic acid, oxidation of, 13, 14 Electron transfer pathway,
Beta carbon, 14–15 in mitochondrial
Beta oxidation, 14–16, 20 membrane, 49–51
biochemical requirements for, 15 Energy homeostasis, 54–57
Blank acid, 39 Entropy, 2
Bomb calorimeter, 4, 5, 8 Enzymes, 12, 57
Exons, 18
Carbohydrate metabolism, 25–32
2-carbon intermediates, energy Fatty acids, oxidation of, 13, 14, 17
extraction, 39–46 First law of thermodynamics, 1–2
Casein, 11, 12 Flavin adenine dinucleotide
Cellular respiration, 19, 20, 25 (FAD), 13
efficiency calculation of, 56 Foods into energy, converting, 11–35
homeostasis of, 55 carbohydrate metabolism, 25–32
substrates for, 54–55 ethical, legal, social implications
Chemical bonds of, 32–35
energy released from, lipid metabolism, 13–20
measuring, 4–5 protein metabolism, 20–25
Chemiosmosis, 52 Free energy, 2
Cinnamic acid, oxidation of, 14
Citrate, 30 Glutamate, 22–23
Citric acid cycle, 43–44, 45, 47 Glutamate dehydrogenase (GDH),
discovery of, 43 22, 23, 24–25
homeostatic regulation of, 44 Glycolysis, 26, 27
Coenzyme A (CoA), 16, 19–20 homeostatic regulation of, 29–30
Coenzymes, 12 GTP, 24–25, 39, 41, 45, 47, 51, 54,
Cofactors, 12–13 56, 61
Covalent bonds, molecular energy
in, 1–9 Homeostasis, 5
Homozygous, 18
Deamination of amino acids, 23–25
Delineation, 41 Introns, 18

[Link]
66 INDEX

Kinase, 30 OMIM, 17
Krebs, Hans Adolf, 39, 40, 42 Oxaloacetic acid, 39–43
Oxidation, 5, 14
Lactase, 25 beta, 14–16, 20
Lactose, 11, 12 of fatty acids, 13, 14, 17
Lehninger, Albert, 16–17, 43–44, 47
Limiting factor, 20 Palmitic acid, 11, 12
Lipid metabolism, 13–20 degradation pathway for, 17
diseases, genetic basis for, 18 Pedigree, 18
limiting factor in, 20 Peptide bonds, 20
Lipids, 20 Phenylacetic acid, oxidation
Long chain acetyl-CoA dehydrogenase of, 13, 14
(LCAD), 19 Phenylbutyric acid, oxidation of, 14
Phenyl isocrontonic acid,
Malnutrition, 58 oxidation of, 14
Mayer, Richard, 33 Phenylproprionic acid, oxidation
Messenger RNA (mRNA), 19 of, 14
Micronutrients, 57 Phosphofructokinase (PFK), 29–31
Mitchell, Peter, 51–52 Protease, 21–22
Mitochondrial matrix, 17, 47 Protein metabolism, 20–25
Mitochondrial membrane, electron Pyruvate, 26
transfer pathway in, 49–51
Molecular energy, in covalent Quick, Armand, 13–14
bonds, 1–9
Multicellular organisms, 56 Redox, 12
Reduced, 5
Negative feedback loop, 30
Nicotinamide adenine dinucleotide Second law of thermodynamics, 2
(NAD+), 5, 13, 24
NADH to, oxidation of, 6 Thermodynamics, 1
Nicotinamide adenine dinucleotide first law of, 1–2
plus hydrogen (NADH), 5, second law of, 2
22, 24, 25, 61
to NAD+, oxidation of, 6 Very long chain acetyl-CoA
Nutrient deficiencies, 58 dehydrogenase (VLCAD),
17–18, 19
Octanoic acid, beta oxidation of, 15
Olestra, 15 Work, 1
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