9 OVERVIEW

Life Is Work
L iving cells require transfusions of energy from outside
sources to perform their many tasks—for example, assembling
polymers, pumping substances across membranes, moving,
and reproducing. The chimpanzee in Figure 9.1 obtains en-
Cellular Respiration ergy for its cells by eating plants; some animals feed on other
organisms that eat plants. The energy stored in the organic

and Fermentation molecules of food ultimately comes from the sun. Energy
flows into an ecosystem as sunlight and exits as heat; in con-
trast, the chemical elements essential to life are recycled
(Figure 9.2). Photosynthesis generates oxygen and organic
molecules used by the mitochondria of eukaryotes (including
plants and algae) as fuel for cellular respiration. Respiration
breaks this fuel down, generating ATP. The waste products of
this type of respiration, carbon dioxide and water, are the raw
materials for photosynthesis. In this chapter, we consider how
cells harvest the chemical energy stored in organic molecules
and use it to generate ATP, the molecule that drives most cel-
lular work. After presenting some basics about respiration, we
will focus on three key pathways of respiration: glycolysis, the
citric acid cycle, and oxidative phosphorylation. We’ll also
consider fermentation, a somewhat simpler pathway coupled
to glycolysis that has deep evolutionary roots.

Light
energy

ECOSYSTEM

 Figure 9.1 How do these leaves power

the work of life for this chimpanzee?
Photosynthesis
in chloroplasts
CO2 + H2O Organic
KEY CONCEPTS + O2
molecules
Cellular respiration
9.1 Catabolic pathways yield energy by oxidizing in mitochondria
organic fuels
9.2 Glycolysis harvests chemical energy by oxidizing
glucose to pyruvate
9.3 After pyruvate is oxidized, the citric acid cycle
completes the energy-yielding oxidation of
organic molecules ATP
ATP powers
most cellular work
9.4 During oxidative phosphorylation, chemiosmosis
couples electron transport to ATP synthesis
Heat
9.5 Fermentation and anaerobic respiration enable energy
cells to produce ATP without the use of oxygen
9.6 Glycolysis and the citric acid cycle connect to  Figure 9.2 Energy flow and chemical recycling in
many other metabolic pathways ecosystems. Energy flows into an ecosystem as sunlight and ultimately
leaves as heat, while the chemical elements essential to life are recycled.
ANIMATION Visit the Study Area at
www.masteringbiology.com for
®
the BioFlix 3-D Animation on
The Carbon Cycle.

CHAPTER 9 Cellular Respiration and Fermentation 163

 This breakdown of glucose is exergonic, having a free-
CONCEPT
9.1 energy change of "686 kcal (2,870 kJ) per mole of glucose
decomposed (∆G # "686 kcal/mol). Recall that a negative ∆G
Catabolic pathways yield energy
indicates that the products of the chemical process store less
by oxidizing organic fuels energy than the reactants and that the reaction can happen
As you learned in Chapter 8, metabolic pathways that release spontaneously—in other words, without an input of energy.
stored energy by breaking down complex molecules are Catabolic pathways do not directly move flagella, pump
called catabolic pathways. Electron transfer plays a major role solutes across membranes, polymerize monomers, or perform
in these pathways. In this section, we consider these other cellular work. Catabolism is linked to work by a chemi-
processes, which are central to cellular respiration. cal drive shaft—ATP, which you learned about in Chapter 8.
To keep working, the cell must regenerate its supply of ATP
Catabolic Pathways and Production of ATP from ADP and P i (see Figure 8.11). To understand how cellu-
Organic compounds possess potential energy as a result of lar respiration accomplishes this, let’s examine the fundamen-
the arrangement of electrons in the bonds between their tal chemical processes known as oxidation and reduction.
atoms. Compounds that can participate in exergonic reac-
Redox Reactions: Oxidation and Reduction
tions can act as fuels. With the help of enzymes, a cell sys-
tematically degrades complex organic molecules that are rich How do the catabolic pathways that decompose glucose and
in potential energy to simpler waste products that have less other organic fuels yield energy? The answer is based on the
energy. Some of the energy taken out of chemical storage can transfer of electrons during the chemical reactions. The relo-
be used to do work; the rest is dissipated as heat. cation of electrons releases energy stored in organic mol-
One catabolic process, fermentation, is a partial degra- ecules, and this energy ultimately is used to synthesize ATP.
dation of sugars or other organic fuel that occurs without the
The Principle of Redox
use of oxygen. However, the most prevalent and efficient
catabolic pathway is aerobic respiration, in which oxygen In many chemical reactions, there is a transfer of one or more
is consumed as a reactant along with the organic fuel (aerobic electrons (e") from one reactant to another. These electron
is from the Greek aer, air, and bios, life). The cells of most eu- transfers are called oxidation-reduction reactions, or redox
karyotic and many prokaryotic organisms can carry out aero- reactions for short. In a redox reaction, the loss of electrons
bic respiration. Some prokaryotes use substances other than from one substance is called oxidation, and the addition of
oxygen as reactants in a similar process that harvests chemi- electrons to another substance is known as reduction. (Note
cal energy without oxygen; this process is called anaerobic res- that adding electrons is called reduction; negatively charged
piration (the prefix an- means “without”). Technically, the electrons added to an atom reduce the amount of positive
term cellular respiration includes both aerobic and anaer- charge of that atom.) To take a simple, nonbiological exam-
obic processes. However, it originated as a synonym for aero- ple, consider the reaction between the elements sodium (Na)
bic respiration because of the relationship of that process to and chlorine (Cl) that forms table salt:
organismal respiration, in which an animal breathes in oxy-
becomes oxidized
gen. Thus, cellular respiration is often used to refer to the aero- (loses electron)
bic process, a practice we follow in most of this chapter. Na + Cl Na+ + Cl–
Although very different in mechanism, aerobic respiration becomes reduced
(gains electron)
is in principle similar to the combustion of gasoline in an au-
tomobile engine after oxygen is mixed with the fuel (hydro- We could generalize a redox reaction this way:
carbons). Food provides the fuel for respiration, and the
becomes oxidized
exhaust is carbon dioxide and water. The overall process can
Xe – + Y X + Ye –
be summarized as follows:
becomes reduced
Organic Carbon
Oxygen S Water Energy
compounds dioxide In the generalized reaction, substance Xe", the electron
donor, is called the reducing agent; it reduces Y, which ac-
Although carbohydrates, fats, and proteins can all be
cepts the donated electron. Substance Y, the electron accep-
processed and consumed as fuel, it is helpful to learn the
tor, is the oxidizing agent; it oxidizes Xe" by removing its
steps of cellular respiration by tracking the degradation of the
electron. Because an electron transfer requires both a donor
sugar glucose (C6H12O6):
and an acceptor, oxidation and reduction always go together.
C6H12O6 6 O2 S 6 CO2 6 H2O Energy (ATP heat) Not all redox reactions involve the complete transfer of
Glucose is the fuel that cells most often use; we will discuss electrons from one substance to another; some change the de-
other organic molecules contained in foods later in the chapter. gree of electron sharing in covalent bonds. The reaction

164 UNIT TWO The Cell

 Reactants Products is respiration: the oxidation of glucose and other molecules
in food. Examine again the summary equation for cellular
becomes oxidized
respiration, but this time think of it as a redox process:
CH4 + 2 O2 CO2 + Energy + 2 H2O becomes oxidized
C6H12O6 + 6 O2 6 CO2 + 6 H2O + Energy
H becomes reduced
becomes reduced

H C H O O O C O H O H
As in the combustion of methane or gasoline, the fuel (glu-
cose) is oxidized and oxygen is reduced. The electrons lose
H potential energy along the way, and energy is released.
Methane Oxygen Carbon dioxide Water In general, organic molecules that have an abundance of
(reducing (oxidizing hydrogen are excellent fuels because their bonds are a source
agent) agent)
of “hilltop” electrons, whose energy may be released as these
electrons “fall” down an energy gradient when they are trans-
 Figure 9.3 Methane combustion as an energy-yielding
redox reaction. The reaction releases energy to the surroundings ferred to oxygen. The summary equation for respiration indi-
because the electrons lose potential energy when they end up being shared cates that hydrogen is transferred from glucose to oxygen. But
unequally, spending more time near electronegative atoms such as oxygen.
the important point, not visible in the summary equation, is
that the energy state of the electron changes as hydrogen
between methane and oxygen, shown in Figure 9.3, is an ex- (with its electron) is transferred to oxygen. In respiration, the
ample. As explained in Chapter 2, the covalent electrons in oxidation of glucose transfers electrons to a lower energy state,
methane are shared nearly equally between the bonded atoms liberating energy that becomes available for ATP synthesis.
because carbon and hydrogen have about the same affinity for The main energy-yielding foods, carbohydrates and fats,
valence electrons; they are about equally electronegative. But are reservoirs of electrons associated with hydrogen. Only the
when methane reacts with oxygen, forming carbon dioxide, barrier of activation energy holds back the flood of electrons
electrons end up shared less equally between the carbon atom to a lower energy state (see Figure 8.12). Without this barrier,
and its new covalent partners, the oxygen atoms, which are a food substance like glucose would combine almost instan-
very electronegative. In effect, the carbon atom has partially taneously with O2. If we supply the activation energy by ig-
“lost” its shared electrons; thus, methane has been oxidized. niting glucose, it burns in air, releasing 686 kcal (2,870 kJ) of
Now let’s examine the fate of the reactant O2. The two heat per mole of glucose (about 180 g). Body temperature is
atoms of the oxygen molecule (O2) share their electrons not high enough to initiate burning, of course. Instead, if
equally. But when oxygen reacts with the hydrogen from you swallow some glucose, enzymes in your cells will lower
methane, forming water, the electrons of the covalent bonds the barrier of activation energy, allowing the sugar to be oxi-
spend more time near the oxygen (see Figure 9.3). In effect, dized in a series of steps.
each oxygen atom has partially “gained” electrons, so the oxy-
gen molecule has been reduced. Because oxygen is so electro-
Stepwise Energy Harvest via NAD
negative, it is one of the most potent of all oxidizing agents.
and the Electron Transport Chain
Energy must be added to pull an electron away from an If energy is released from a fuel all at once, it cannot be har-
atom, just as energy is required to push a ball uphill. The nessed efficiently for constructive work. For example, if a
more electronegative the atom (the stronger its pull on elec- gasoline tank explodes, it cannot drive a car very far. Cellular
trons), the more energy is required to take an electron away respiration does not oxidize glucose in a single explosive step
from it. An electron loses potential energy when it shifts either. Rather, glucose and other organic fuels are broken
from a less electronegative atom toward a more electronega- down in a series of steps, each one catalyzed by an enzyme.
tive one, just as a ball loses potential energy when it rolls At key steps, electrons are stripped from the glucose. As is
downhill. A redox reaction that moves electrons closer to often the case in oxidation reactions, each electron travels
oxygen, such as the burning (oxidation) of methane, there- with a proton—thus, as a hydrogen atom. The hydrogen
fore releases chemical energy that can be put to work. atoms are not transferred directly to oxygen, but instead are
usually passed first to an electron carrier, a coenzyme called
Oxidation of Organic Fuel Molecules NAD (nicotinamide adenine dinucleotide, a derivative of
During Cellular Respiration the vitamin niacin). NAD is well suited as an electron carrier
The oxidation of methane by oxygen is the main combustion because it can cycle easily between oxidized (NAD ) and re-
reaction that occurs at the burner of a gas stove. The combus- duced (NADH) states. As an electron acceptor, NAD func-
tion of gasoline in an automobile engine is also a redox reac- tions as an oxidizing agent during respiration.
tion; the energy released pushes the pistons. But the How does NAD trap electrons from glucose and other or-
energy-yielding redox process of greatest interest to biologists ganic molecules? Enzymes called dehydrogenases remove a

CHAPTER 9 Cellular Respiration and Fermentation 165

 2 e– + 2 H+
2 e – + H+
NAD+ NADH H+
Dehydrogenase
H O O
Reduction of NAD+ H H
C NH2 + 2[H] C NH2 + H+
(from food) Oxidation of NADH
N+ Nicotinamide N Nicotinamide
(oxidized form) (reduced form)
O CH2
O
O P O–

O H H
O P O– HO OH
NH2
O CH2  Figure 9.4 NAD as an electron shuttle. The full name for
N
N NAD , nicotinamide adenine dinucleotide, describes its structure: The
H
molecule consists of two nucleotides joined together at their phosphate
N N H
O groups (shown in yellow). (Nicotinamide is a nitrogenous base, although
not one that is present in DNA or RNA; see Figure 5.26.) The enzymatic
transfer of 2 electrons and 1 proton (H ) from an organic molecule in food
H H to NAD reduces the NAD to NADH; the second proton (H ) is released.
HO OH Most of the electrons removed from food are transferred initially to NAD .

pair of hydrogen atoms (2 electrons and 2 protons) from the gases combine explosively. In fact, combustion of liquid H2
substrate (glucose, in this example), thereby oxidizing it. The and O2 is harnessed to power the main engines of the space
enzyme delivers the 2 electrons along with 1 proton to its shuttle after it is launched, boosting it into orbit. The explo-
coenzyme, NAD (Figure 9.4). The other proton is released sion represents a release of energy as the electrons of hydrogen
as a hydrogen ion (H ) into the surrounding solution: “fall” closer to the electronegative oxygen atoms. Cellular res-
piration also brings hydrogen and oxygen together to form
Dehydrogenase
H C OH + NAD+ C O + NADH + H+ water, but there are two important differences. First, in cellular
respiration, the hydrogen that reacts with oxygen is derived
By receiving 2 negatively charged electrons but only 1 posi- from organic molecules rather than H2. Second, instead of
tively charged proton, NAD has its charge neutralized when occurring in one explosive reaction, respiration uses an
it is reduced to NADH. The name NADH
shows the hydrogen that has been re- H2 + 1/2 O2 + 1
/2 O2
2H
ceived in the reaction. NAD is the (from food via NADH)
most versatile electron acceptor in cellu- Controlled
lar respiration and functions in several release of
+
2H + 2e –
of the redox steps during the break- energy for
synthesis of
down of glucose. ATP
Elec

ATP
Electrons lose very little of their po-
Free energy, G

Free energy, G

tron ain

tential energy when they are transferred Explosive ATP

ch

release of
from glucose to NAD . Each NADH
tran

heat and light ATP

molecule formed during respiration energy
spor

represents stored energy that can be

2 e–
t

tapped to make ATP when the electrons

12 O
complete their “fall” down an energy 2 H+ 2

gradient from NADH to oxygen.

How do electrons that are extracted H2O H2O
from glucose and stored as potential en-
ergy in NADH finally reach oxygen? It (a) Uncontrolled reaction (b) Cellular respiration
will help to compare the redox chem-
istry of cellular respiration to a much  Figure 9.5 An introduction to electron transport chains. (a) The one-step exergonic
simpler reaction: the reaction between reaction of hydrogen with oxygen to form water releases a large amount of energy in the form of
heat and light: an explosion. (b) In cellular respiration, the same reaction occurs in stages: An
hydrogen and oxygen to form water
electron transport chain breaks the “fall” of electrons in this reaction into a series of smaller steps
(Figure 9.5a). Mix H2 and O2, provide a and stores some of the released energy in a form that can be used to make ATP. (The rest of the
spark for activation energy, and the energy is released as heat.)

166 UNIT TWO The Cell

electron transport chain to break the fall of electrons to oxygen The Stages of Cellular Respiration: A Preview
into several energy-releasing steps (Figure 9.5b). An electron
The harvesting of energy from glucose by cellular respiration
transport chain consists of a number of molecules, mostly
is a cumulative function of three metabolic stages:
proteins, built into the inner membrane of the mitochondria of
eukaryotic cells and the plasma membrane of aerobically 1. Glycolysis (color-coded teal throughout the chapter)
respiring prokaryotes. Electrons removed from glucose are 2. Pyruvate oxidation and the citric acid cycle
shuttled by NADH to the “top,” higher-energy end of the (color-coded salmon)
chain. At the “bottom,” lower-energy end, O2 captures these 3. Oxidative phosphorylation: electron transport and
electrons along with hydrogen nuclei (H ), forming water. chemiosmosis (color-coded violet)
Electron transfer from NADH to oxygen is an exergonic re-
action with a free-energy change of "53 kcal/mol ("222 Biochemists usually reserve the term cellular respiration for
kJ/mol). Instead of this energy being released and wasted in a stages 2 and 3. We include glycolysis, however, because most
single explosive step, electrons cascade down the chain from respiring cells deriving energy from glucose use glycolysis to
one carrier molecule to the next in a series of redox reactions, produce the starting material for the citric acid cycle.
losing a small amount of energy with each step until they fi- As diagrammed in Figure 9.6, glycolysis and pyruvate oxida-
nally reach oxygen, the terminal electron acceptor, which has tion followed by the citric acid cycle are the catabolic pathways
a very great affinity for electrons. Each “downhill” carrier is that break down glucose and other organic fuels. Glycolysis,
more electronegative than, and thus capable of oxidizing, its which occurs in the cytosol, begins the degradation process by
“uphill” neighbor, with oxygen at the bottom of the chain. breaking glucose into two molecules of a compound called
Therefore, the electrons removed from glucose by NAD fall pyruvate. In eukaryotes, pyruvate enters the mitochondrion
down an energy gradient in the electron transport chain to a and is oxidized to a compound called acetyl CoA, which enters
far more stable location in the electronegative oxygen atom. the citric acid cycle. There, the breakdown of glucose to car-
Put another way, oxygen pulls electrons down the chain in an bon dioxide is completed. (In prokaryotes, these processes take
energy-yielding tumble analogous to gravity pulling objects place in the cytosol.) Thus, the carbon dioxide produced by res-
downhill. piration represents fragments of oxidized organic molecules.
In summary, during cellular respiration, most electrons Some of the steps of glycolysis and the citric acid cycle are
travel the following “downhill” route: glucose → NADH → redox reactions in which dehydrogenases transfer electrons
electron transport chain → oxygen. Later in this chapter, you from substrates to NAD , forming NADH. In the third stage of
will learn more about how the cell uses the energy released respiration, the electron transport chain accepts electrons from
from this exergonic electron fall to regenerate its supply of the breakdown products of the first two stages (most often via
ATP. For now, having covered the basic redox mechanisms of NADH) and passes these electrons from one molecule to an-
cellular respiration, let’s look at the entire process by which other. At the end of the chain, the electrons are combined with
energy is harvested from organic fuels. molecular oxygen and hydrogen ions (H ), forming water (see

 Figure 9.6 An overview of cellular

respiration. During glycolysis, each glucose
molecule is broken down into two molecules of Electrons carried
Electrons
the compound pyruvate. In eukaryotic cells, as via NADH and
carried
shown here, the pyruvate enters the via NADH FADH2
mitochondrion. There it is oxidized to acetyl
CoA, which is further oxidized to CO2 in the
citric acid cycle. NADH and a similar electron
carrier, a coenzyme called FADH2, transfer Pyruvate Oxidative
Glycolysis oxidation Citric phosphorylation:
electrons derived from glucose to electron
acid electron transport
transport chains, which are built into the inner Glucose Pyruvate Acetyl CoA cycle and
mitochondrial membrane. (In prokaryotes, the chemiosmosis
electron transport chains are located in the
plasma membrane.) During oxidative
phosphorylation, electron transport chains
convert the chemical energy to a form used for CYTOSOL MITOCHONDRION
ATP synthesis in the process called chemiosmosis.

ANIMATION Visit the Study Area at ATP ATP ATP

www.masteringbiology.com for
the BioFlix® 3-D Animation on Substrate-level Substrate-level Oxidative
Cellular Respiration. phosphorylation phosphorylation phosphorylation

CHAPTER 9 Cellular Respiration and Fermentation 167

Figure 9.5b). The energy released at each step of the chain is
stored in a form the mitochondrion (or prokaryotic cell) can
CONCEPT
9.2
use to make ATP from ADP. This mode of ATP synthesis is
Glycolysis harvests chemical energy
called oxidative phosphorylation because it is powered by
the redox reactions of the electron transport chain. by oxidizing glucose to pyruvate
In eukaryotic cells, the inner membrane of the mitochon-
The word glycolysis means “sugar splitting,” and that is exactly
drion is the site of electron transport and chemiosmosis, the
what happens during this pathway. Glucose, a six-carbon
processes that together constitute oxidative phosphorylation.
sugar, is split into two three-carbon sugars. These smaller sug-
(In prokaryotes, these processes take place in the plasma
ars are then oxidized and their remaining atoms rearranged
membrane.) Oxidative phosphorylation accounts for almost
to form two molecules of pyruvate. (Pyruvate is the ionized
90% of the ATP generated by respiration. A smaller amount
form of pyruvic acid.)
of ATP is formed directly in a few reactions of glycolysis and
As summarized in Figure 9.8, glycolysis can be divided into
the citric acid cycle by a mechanism called substrate-level
two phases: energy investment and energy payoff. During the
phosphorylation (Figure 9.7). This mode of ATP synthesis
energy investment phase, the cell actually spends ATP. This
occurs when an enzyme transfers a phosphate group from a
investment is repaid with interest during the energy payoff
substrate molecule to ADP, rather than adding an inorganic
phase, when ATP is produced by substrate-level phosphoryla-
phosphate to ADP as in oxidative phosphorylation. “Sub-
tion and NAD is reduced to NADH by electrons released
strate molecule” here refers to an organic molecule generated
from the oxidation of glucose. The net energy yield from gly-
as an intermediate during the catabolism of glucose.
colysis, per glucose molecule, is 2 ATP plus 2 NADH. The ten
For each molecule of glucose degraded to carbon dioxide
steps of the glycolytic pathway are shown in Figure 9.9.
and water by respiration, the cell makes up to about 32 mol-
All of the carbon originally present in glucose is accounted
ecules of ATP, each with 7.3 kcal/mol of free energy. Respira-
for in the two molecules of pyruvate; no carbon is released as
tion cashes in the large denomination of energy banked in a
CO2 during glycolysis. Glycolysis occurs whether or not O2 is
single molecule of glucose (686 kcal/mol) for the small
present. However, if O2 is present, the chemical energy stored
change of many molecules of ATP, which is more practical for
in pyruvate and NADH can be extracted by pyruvate oxida-
the cell to spend on its work.
tion, the citric acid
This preview has introduced you to how glycolysis, the
cycle, and oxidative
citric acid cycle, and oxidative phosphorylation fit into the
phosphorylation.
process of cellular respiration. We are now ready to take a Citric Oxidative
Glycolysis Pyruvate acid phosphorylation
closer look at each of these three stages of respiration. oxidation cycle

CONCEPT CHECK 9.1

ATP ATP ATP
1. Compare and contrast aerobic and anaerobic Energy Investment Phase
respiration. Glucose
2. WHAT IF? If the following redox reaction occurred,
which compound would be oxidized? Which reduced?
2 ADP + 2 P 2 ATP used
C4H6O5 NAD → C4H4O5 NADH H
For suggested answers, see Appendix A.

Energy Payoff Phase

4 ADP + 4 P 4 ATP formed

Enzyme Enzyme

ADP 2 NAD+ + 4 e– + 4 H+ 2 NADH + 2 H+

P
Substrate + ATP 2 Pyruvate + 2 H2O

Product
Net
 Figure 9.7 Substrate-level phosphorylation. Some ATP is made Glucose 2 Pyruvate + 2 H2O
by direct transfer of a phosphate group from an organic substrate to ADP 4 ATP formed – 2 ATP used 2 ATP
by an enzyme. (For examples in glycolysis, see Figure 9.9, steps 7 and 10.)
MAKE CONNECTIONS Review Figure 8.8 on page 149. Do you think 2 NAD+ + 4 e– + 4 H+ 2 NADH + 2 H+
the potential energy is higher for the reactants or the products in the
reaction shown above? Explain.  Figure 9.8 The energy input and output of glycolysis.

168 UNIT TWO The Cell

 Figure 9.9 A closer look at glycolysis. The orientation diagram on the left relates
glycolysis to the entire process of respiration. Note that glycolysis is a source of ATP and NADH.
Citric WHAT IF? What would happen if you removed the dihydroxyacetone phosphate generated in
Pyruvate Oxidative
Glycolysis acid
oxidation cycle
phosphorylation step 4 as fast as it was produced?

ATP ATP ATP Glycolysis: Energy Investment Phase

ATP ATP
Glucose Glucose 6-phosphate Fructose 6-phosphate Fructose 1,6-bisphosphate
CH2OH ADP CH O P CH2O P CH2OH ADP P O CH2 CH2 O P Aldolase
2
O H O O O cleaves the
H H H H H
H HO H HO sugar molecule
OH H OH H H H OH
HO OH Hexokinase HO OH Phosphogluco-
OH
Phospho- into two different
H OH H OH isomerase HO H fructokinase HO H three-carbon
1 Aldolase 4 sugars (isomers).
2 3

Hexokinase Glucose 6- Phosphofructokinase

transfers a phosphate is transfers a phosphate
phosphate group converted to its group from ATP to the Dihydroxyacetone Glyceraldehyde
from ATP to isomer, fructose opposite end of the phosphate 3-phosphate
glucose, making it 6-phosphate. sugar, investing a second P O CH2 H C O
more chemically molecule of ATP. This is C O CHOH
reactive. The charge a key step for regulation
on the phosphate also of glycolysis. CH2OH CH2 O P
To step 6
traps the sugar in the cell. Isomerase 5
Isomerase catalyzes the reversible conversion between the two isomers.
This reaction never reaches equilibrium: Glyceraldehyde 3-phosphate is
used as the substrate of the next reaction (step 6) as fast as it forms.

The energy payoff phase occurs after glucose is

split into two three-carbon sugars. Thus, the Glycolysis: Energy Payoff Phase
coefficient 2 precedes all molecules in this phase.

2 ATP 2 ATP
2 NADH 2 H2O
2 ADP
2 NAD + + 2 H+ 2 ADP 2 2 2 2
O– O– O– O–
2
P O C O C O C O C O C O

CHOH CHOH H C O P C O P C O
Triose Phospho- Phospho- Enolase Pyruvate
phosphate 2 Pi CH2 O P glycerokinase CH2 O P glyceromutase CH2OH CH2 kinase CH3
dehydrogenase 9
1,3-Bisphospho- 7 3-Phospho- 8 2-Phospho- Phosphoenol- 10 Pyruvate
6 glycerate glycerate glycerate pyruvate (PEP)

This enzyme catalyzes two The phosphate group added This enzyme Enolase causes The phosphate
sequential reactions. First, the in the previous step is relocates the a double bond to group is transferred
sugar is oxidized by the transferred to ADP (substrate- remaining form in the substrate from PEP to ADP
transfer of electrons to NAD+, level phosphorylation) in an phosphate by extracting a water (a second example
forming NADH. Second, the exergonic reaction. The group. molecule, yielding of substrate-level
energy released from this carbonyl group of a sugar phosphoenolpyruvate phosphorylation),
exergonic redox reaction is has been oxidized to the (PEP), a compound forming pyruvate.
used to attach a phosphate carboxyl group ( —COO–) of with a very high
group to the oxidized an organic acid potential energy.
substrate, making a product (3-phosphoglycerate).
of very high potential energy.

CONCEPT CHECK 9.2

1. During the redox reaction in glycolysis (step 6 in and related molecules (see Concept 8.5, p. 158). Consid-
Figure 9.9), which molecule acts as the oxidizing ering the overall result of glycolysis, would you expect
agent? The reducing agent? ATP to inhibit or stimulate activity of this enzyme?
2. MAKE CONNECTIONS Step 3 in Figure 9.9 is a major (Hint: Make sure you consider the role of ATP as an al-
point of regulation of glycolysis. The enzyme losteric regulator, not as a substrate of the enzyme.)
phosphofructokinase is allosterically regulated by ATP For suggested answers, see Appendix A.

CHAPTER 9 Cellular Respiration and Fermentation 169

 CONCEPT
9.3 storing energy in the form of NADH. 3 Finally, coenzyme A
(CoA), a sulfur-containing compound derived from a B vita-
After pyruvate is oxidized, min, is attached via its sulfur atom to the acetate, forming
acetyl CoA, which has a high potential energy; in other
the citric acid cycle completes words, the reaction of acetyl CoA to yield lower-energy prod-
the energy-yielding oxidation ucts is highly exergonic. This molecule will now feed its acetyl
of organic molecules group into the citric acid cycle for further oxidation.

Glycolysis releases less than a quarter of the chemical energy The Citric Acid Cycle
in glucose that can be released by cells; most of the energy re-
The citric acid cycle is also called the tricarboxylic acid cycle
mains stockpiled in the two molecules of pyruvate. If molecu-
or the Krebs cycle, the latter honoring Hans Krebs, the
lar oxygen is present, the pyruvate enters a mitochondrion (in
German-British scientist who was largely responsible for
eukaryotic cells), where the oxidation of glucose is completed.
working out the pathway in the 1930s. The cycle functions as
(In prokaryotic cells, this process occurs in the cytosol.)
a metabolic furnace that oxidizes organic fuel derived from
pyruvate. Figure 9.11 summarizes the inputs and outputs as
Oxidation of Pyruvate to Acetyl CoA
pyruvate is broken down to three CO2 molecules, including
Upon entering the mitochondrion via active transport, pyru- the molecule of CO2 released during the conversion of pyru-
vate is first converted to a compound called acetyl coenzyme vate to acetyl CoA. The cycle generates 1 ATP per turn by
A, or acetyl CoA (Figure 9.10). This step, linking glycolysis
and the citric acid cycle, is carried out by a multienzyme com-
plex that catalyzes three reactions: 1 Pyruvate’s carboxyl
group (—COO"), which is already fully oxidized and thus has Pyruvate
Citric Oxidative
Glycolysis acid phosphorylation
oxidation
little chemical energy, is removed and given off as a molecule cycle
Pyruvate
of CO2. (This is the first step in which CO2 is released during (from glycolysis,
respiration.) 2 The remaining two-carbon fragment is 2 molecules per glucose)
ATP ATP ATP
oxidized, forming acetate (CH3COO", the ionized form of
acetic acid). The extracted electrons are transferred to NAD ,
CO2
NAD+
CoA
NADH
+ H+
Pyruvate
Citric Oxidative Acetyl CoA
Glycolysis acid phosphorylation
oxidation cycle CoA

CoA
ATP ATP ATP
MITOCHONDRION
CYTOSOL CO2 Coenzyme A
O–
1 3 S-CoA
C O Citric
C O acid
C O 2 cycle 2 CO2
CH3
CH3
NAD + NADH + H + Acetyl CoA FADH2 3 NAD+
Pyruvate

Transport protein FAD 3 NADH

+ 3 H+
 Figure 9.10 Oxidation of pyruvate to acetyl CoA, the ADP + P i
step before the citric acid cycle. Pyruvate is a charged molecule,
so in eukaryotic cells it must enter the mitochondrion via active ATP
transport, with the help of a transport protein. Next, a complex of
several enzymes (the pyruvate dehydrogenase complex) catalyzes the
three numbered steps, which are described in the text. The acetyl group  Figure 9.11 An overview of pyruvate oxidation and the
of acetyl CoA will enter the citric acid cycle. The CO2 molecule will citric acid cycle. The inputs and outputs per pyruvate molecule are
diffuse out of the cell. By convention, coenzyme A is abbreviated S-CoA shown. To calculate on a per-glucose basis, multiply by 2, because each
when it is attached to a molecule, emphasizing the sulfur atom (S). glucose molecule is split during glycolysis into two pyruvate molecules.

170 UNIT TWO The Cell

substrate-level phosphorylation, but most of the chemical Now let’s look at the citric acid cycle in more detail. The
energy is transferred to NAD and a related electron carrier, cycle has eight steps, each catalyzed by a specific enzyme.
the coenzyme FAD (flavin adenine dinucleotide, derived You can see in Figure 9.12 that for each turn of the citric acid
from riboflavin, a B vitamin), during the redox reactions. The cycle, two carbons (red) enter in the relatively reduced form
reduced coenzymes, NADH and FADH2, shuttle their cargo of of an acetyl group (step 1), and two different carbons (blue)
high-energy electrons into the electron transport chain. leave in the completely oxidized form of CO2 molecules

Citric Oxidative
Glycolysis Pyruvate acid
oxidation phosphorylation
cycle

S-CoA 1 Acetyl CoA (from

oxidation of pyruvate) 2 Citrate is
C O
ATP ATP ATP adds its two-carbon acetyl converted to
CH3 group to oxaloacetate, its isomer,
producing citrate. isocitrate, by
Acetyl CoA
removal of
CoA-SH one water
8 The substrate molecule and
is oxidized, addition of
reducing NAD+ to O C COO– another.
NADH
NADH and
+ H+ CH2 1 COO– H2O
regenerating
–
oxaloacetate. NAD +
COO CH2 COO–

8 Oxaloacetate HO C COO –
CH2
2
CH2 HC COO–
COO–
COO– HO CH
HO CH
Malate Citrate COO–
CH2 3 Isocitrate
–
Isocitrate is oxidized,
COO
7 Addition of NAD + reducing
a water NAD+ to
Citric NADH NADH. Then
molecule 3
acid + H+ the resulting
rearranges 7 cycle
bonds in the H2O compound
CO2 loses a CO2
substrate. COO–
COO– molecule.
CH
Fumarate CH2
CoA-SH
HC
CH2 α-Ketoglutarate
COO–
C O
4
6 CoA-SH COO–
COO– COO–

CH2 5 CH2 4 Another CO2

FADH 2
CH2 CH2 CO2 is lost, and the
NAD + resulting
FAD COO– C O
6 Two compound is
Succinate S-CoA NADH oxidized,
hydrogens are Pi
transferred to reducing NAD+
GTP GDP Succinyl + H+
FAD, forming to NADH.
CoA The remain-
FADH2 and
ing molecule is
oxidizing ADP then attached
succinate. 5 CoA is displaced by a phosphate group,
to coenzyme A
which is transferred to GDP, forming GTP, a
ATP by an unstable
molecule with functions similar to ATP. GTP
bond.
can also be used, as shown, to generate ATP.

 Figure 9.12 A closer look at the citric from each other.) Notice that the carbon atoms around. In eukaryotic cells, all the citric acid cycle
acid cycle. In the chemical structures, red type that enter the cycle from acetyl CoA do not leave enzymes are located in the mitochondrial matrix
traces the fate of the two carbon atoms that enter the cycle in the same turn. They remain in the except for the enzyme that catalyzes step 6, which
the cycle via acetyl CoA (step 1), and blue type cycle, occupying a different location in the resides in the inner mitochondrial membrane.
indicates the two carbons that exit the cycle as molecules on their next turn, after another acetyl Carboxylic acids are represented in their ionized
CO2 in steps 3 and 4. (The red labeling goes only group is added. As a consequence, the forms, as —COO , because the ionized forms
through step 5 because the succinate molecule is oxaloacetate that is regenerated at step 8 is prevail at the pH within the mitochondrion. For
symmetrical; the two ends cannot be distinguished composed of different carbon atoms each time example, citrate is the ionized form of citric acid.

CHAPTER 9 Cellular Respiration and Fermentation 171

(steps 3 and 4). The acetyl group of acetyl CoA joins the cycle phosphorylation: 2 net ATP from glycolysis and 2 ATP from
by combining with the compound oxaloacetate, forming cit- the citric acid cycle. At this point, molecules of NADH (and
rate (step 1). (Citrate is the ionized form of citric acid, for FADH2) account for most of the energy extracted from the glu-
which the cycle is named.) The next seven steps decompose cose. These electron escorts link glycolysis and the citric acid
the citrate back to oxaloacetate. It is this regeneration of ox- cycle to the machinery of oxidative phosphorylation, which
aloacetate that makes this process a cycle. uses energy released by the electron transport chain to power
Now let’s tally the energy-rich molecules produced by the ATP synthesis. In this section, you will learn first how the elec-
citric acid cycle. For each acetyl group entering the cycle, 3 tron transport chain works and then how electron flow down
NAD! are reduced to NADH (steps 3, 4, and 8). In step 6, the chain is coupled to ATP synthesis.
electrons are transferred not to NAD!, but to FAD, which ac-
cepts 2 electrons and 2 protons to become FADH2. In many
The Pathway of Electron Transport
animal tissue cells, step 5 produces a guanosine triphosphate
(GTP) molecule by substrate-level phosphorylation, as shown The electron transport chain is a collection of molecules em-
in Figure 9.12. GTP is a molecule similar to ATP in its struc- bedded in the inner membrane of the mitochondrion in eu-
ture and cellular function. This GTP may be used to make an karyotic cells. (In prokaryotes, these molecules reside in the
ATP molecule (as shown) or directly power work in the cell. plasma membrane.) The folding of the inner membrane to
In the cells of plants, bacteria, and some animal tissues, step form cristae increases its surface area, providing space for
5 forms an ATP molecule directly by substrate-level phospho- thousands of copies of the chain in each mitochondrion.
rylation. The output from step 5 represents the only ATP gen- (Once again, we see that structure fits function.) Most com-
erated during the citric acid cycle. ponents of the chain are proteins, which exist in multi-
Most of the ATP produced by respiration results from ox- protein complexes numbered I through IV. Tightly bound to
idative phosphorylation, when the NADH and FADH2 pro- these proteins are prosthetic groups, nonprotein components
duced by the citric acid cycle relay the electrons extracted essential for the catalytic functions of certain enzymes.
from food to the electron transport chain. In the process, Figure 9.13 shows the sequence of electron carriers in the
they supply the necessary energy for the phosphorylation of electron transport chain and the drop in free energy as elec-
ADP to ATP. We will explore this process in the next section. trons travel down the chain. During electron transport along
the chain, electron carriers alternate between reduced and
CONCEPT CHECK 9.3 oxidized states as they accept and donate electrons. Each
component of the chain becomes reduced when it accepts
1. Name the molecules that conserve most of the energy
electrons from its “uphill” neighbor, which has a lower affin-
from the citric acid cycle’s redox reactions. How is
ity for electrons (is less electronegative). It then returns to its
this energy converted to a form that can be used to
oxidized form as it passes electrons to its “downhill,” more
make ATP?
electronegative neighbor.
2. What processes in your cells produce the CO2 that
Now let’s take a closer look at the electron transport chain
you exhale?
in Figure 9.13. We’ll first describe the passage of electrons
3. WHAT IF? The conversions shown in Figure 9.10
through complex I in some detail, as an illustration of the
and step 4 of Figure 9.12 are each catalyzed by a large
general principles involved in electron transport. Electrons
multienzyme complex. What similarities are there in
removed from glucose by NAD!, during glycolysis and the
the reactions that occur in these two cases?
citric acid cycle, are transferred from NADH to the first mol-
For suggested answers, see Appendix A.
ecule of the electron transport chain in complex I. This mol-
ecule is a flavoprotein, so named because it has a prosthetic
group called flavin mononucleotide (FMN). In the next redox
CONCEPT
9.4 reaction, the flavoprotein returns to its oxidized form as it
passes electrons to an iron-sulfur protein (Fe·S in complex I),
During oxidative phosphorylation, one of a family of proteins with both iron and sulfur tightly
chemiosmosis couples electron bound. The iron-sulfur protein then passes the electrons to a
transport to ATP synthesis compound called ubiquinone (Q in Figure 9.13). This elec-
tron carrier is a small hydrophobic molecule, the only mem-
Our main objective in this chapter is to learn how cells harvest ber of the electron transport chain that is not a protein.
the energy of glucose and other nutrients in food to make ATP. Ubiquinone is individually mobile within the membrane
But the metabolic components of respiration we have dis- rather than residing in a particular complex. (Another name
sected so far, glycolysis and the citric acid cycle, produce only for ubiquinone is coenzyme Q, or CoQ; you may have seen it
4 ATP molecules per glucose molecule, all by substrate-level sold as a nutritional supplement.)

172 UNIT TWO The Cell

 each a different protein with a slightly different electron-
carrying heme group. The last cytochrome of the chain, cyt a3,
Oxidative
Pyruvate
Citric
phosphorylation: passes its electrons to oxygen, which is very electronegative.
Glycolysis acid
oxidation electron transport
cycle
and chemiosmosis Each oxygen atom also picks up a pair of hydrogen ions from
the aqueous solution, forming water.
Another source of electrons for the transport chain is
ATP ATP ATP
FADH2, the other reduced product of the citric acid cycle. No-
tice in Figure 9.13 that FADH2 adds its electrons to the elec-
NADH
tron transport chain from within complex II, at a lower
50 energy level than NADH does. Consequently, although
2 e– NADH and FADH2 each donate an equivalent number of
NAD+
FADH2 electrons (2) for oxygen reduction, the electron transport
2 e–
chain provides about one-third less energy for ATP synthesis
FAD Multiprotein
I complexes
when the electron donor is FADH2 rather than NADH. We’ll
40 FMN
II see why in the next section.
Fe•S Fe•S
The electron transport chain makes no ATP directly. In-
Free energy (G) relative to O2 (kcal/mol)

Q
III stead, it eases the fall of electrons from food to oxygen,
Cyt b breaking a large free-energy drop into a series of smaller steps
30 Fe•S that release energy in manageable amounts. How does the
Cyt c1 IV mitochondrion (or the prokaryotic plasma membrane) cou-
Cyt c ple this electron transport and energy release to ATP synthe-
Cyt a sis? The answer is a mechanism called chemiosmosis.
Cyt a3
20
Chemiosmosis: The Energy-Coupling
Mechanism
Populating the inner membrane of the mitochondrion or the
2 e– prokaryotic plasma membrane are many copies of a protein
10
(originally from complex called ATP synthase, the enzyme that actually
NADH or FADH2) makes ATP from ADP and inorganic phosphate. ATP synthase
works like an ion pump running in reverse. Recall from
Chapter 7 that ion pumps usually use ATP as an energy
0 2 H+ + 12 O2 source to transport ions against their gradients. In fact, the
proton pump shown in Figure 7.20 is an ATP synthase. As we
mentioned in Chapter 8, enzymes can catalyze a reaction in
either direction, depending on the ∆G for the reaction, which
H2O
is affected by the local concentrations of reactants and prod-
 Figure 9.13 Free-energy change during electron ucts. Rather than hydrolyzing ATP to pump protons against
transport. The overall energy drop (∆G) for electrons traveling from their concentration gradient, under the conditions of cellular
NADH to oxygen is 53 kcal/mol, but this “fall” is broken up into a series respiration ATP synthase uses the energy of an existing ion
of smaller steps by the electron transport chain. (An oxygen atom is
represented here as 1⁄2 O2 to emphasize that the electron transport gradient to power ATP synthesis. The power source for the
chain reduces molecular oxygen, O2, not individual oxygen atoms.) ATP synthase is a difference in the concentration of H! on
opposite sides of the inner mitochondrial membrane. (We
can also think of this gradient as a difference in pH, since pH
is a measure of H! concentration.) This process, in which en-
ergy stored in the form of a hydrogen ion gradient across a
Most of the remaining electron carriers between membrane is used to drive cellular work such as the synthesis
ubiquinone and oxygen are proteins called cytochromes. of ATP, is called chemiosmosis (from the Greek osmos,
Their prosthetic group, called a heme group, has an iron atom push). We have previously used the word osmosis in dis-
that accepts and donates electrons. (It is similar to the heme cussing water transport, but here it refers to the flow of H!
group in hemoglobin, the protein of red blood cells, except across a membrane.
that the iron in hemoglobin carries oxygen, not electrons.) From studying the structure of ATP synthase, scientists
The electron transport chain has several types of cytochromes, have learned how the flow of H! through this large enzyme

CHAPTER 9 Cellular Respiration and Fermentation 173

powers ATP generation. ATP synthase is a multisubunit com- its mitochondrial location in Figure 9.15. The chain is an
plex with four main parts, each made up of multiple energy converter that uses the exergonic flow of electrons
polypeptides. Protons move one by one into binding sites on from NADH and FADH2 to pump H! across the membrane,
one of the parts (the rotor), causing it to spin in a way that from the mitochondrial matrix into the intermembrane
catalyzes ATP production from ADP and inorganic phosphate space. The H! has a tendency to move back across the mem-
(Figure 9.14). The flow of protons thus behaves somewhat brane, diffusing down its gradient. And the ATP synthases
like a rushing stream that turns a waterwheel. ATP synthase is are the only sites that provide a route through the mem-
the smallest molecular rotary motor known in nature. brane for H!. As we described previously, the passage of H!
How does the inner mitochondrial membrane or the through ATP synthase uses the exergonic flow of H! to drive
prokaryotic plasma membrane generate and maintain the the phosphorylation of ADP. Thus, the energy stored in an
H! gradient that drives ATP synthesis by the ATP synthase H! gradient across a membrane couples the redox reactions
protein complex? Establishing the H! gradient is a major of the electron transport chain to ATP synthesis, an example
function of the electron transport chain, which is shown in of chemiosmosis.
At this point, you may be wondering how the electron
transport chain pumps hydrogen ions. Researchers have
INTERMEMBRANE SPACE
found that certain members of the electron transport chain
1 H+ ions flowing accept and release protons (H!) along with electrons. (The
down their gradient aqueous solutions inside and surrounding the cell are a ready
enter a half channel in
a stator, which is source of H!.) At certain steps along the chain, electron
anchored in the transfers cause H! to be taken up and released into the sur-
membrane. rounding solution. In eukaryotic cells, the electron carriers
are spatially arranged in the inner mitochondrial membrane
2 H+ ions enter binding in such a way that H! is accepted from the mitochondrial
sites within a rotor, matrix and deposited in the intermembrane space (see Figure
H+ changing the shape of
Stator each subunit so that 9.15). The H! gradient that results is referred to as a proton-
Rotor the rotor spins within motive force, emphasizing the capacity of the gradient to
the membrane. perform work. The force drives H! back across the membrane
through the H! channels provided by ATP synthases.
3 Each H+ ion makes one In general terms, chemiosmosis is an energy-coupling mecha-
complete turn before nism that uses energy stored in the form of an H! gradient across
leaving the rotor and
passing through a second a membrane to drive cellular work. In mitochondria, the energy
half channel in the stator for gradient formation comes from exergonic redox reac-
into the mitochondrial tions, and ATP synthesis is the work performed. But chemi-
matrix.
Internal osmosis also occurs elsewhere and in other variations.
rod Chloroplasts use chemiosmosis to generate ATP during
4 Spinning of the
rotor causes an internal photosynthesis; in these organelles, light (rather than chemi-
Catalytic rod to spin as well. This cal energy) drives both electron flow down an electron trans-
knob rod extends like a stalk port chain and the resulting H! gradient formation.
into the knob below it,
which is held stationary Prokaryotes, as already mentioned, generate H! gradients
ADP by part of the stator. across their plasma membranes. They then tap the proton-
+ motive force not only to make ATP inside the cell but also to
Pi ATP 5 Turning of the rod rotate their flagella and to pump nutrients and waste prod-
activates catalytic sites
in the knob that ucts across the membrane. Because of its central importance
produce ATP from ADP to energy conversions in prokaryotes and eukaryotes,
and P i . chemiosmosis has helped unify the study of bioenergetics.
MITOCHONDRIAL MATRIX Peter Mitchell was awarded the Nobel Prize in 1978 for origi-
nally proposing the chemiosmotic model.

 Figure 9.14 ATP synthase, a molecular mill. The ATP

synthase protein complex functions as a mill, powered by the flow of
An Accounting of ATP Production
hydrogen ions. Multiple copies of this complex reside in mitochondrial by Cellular Respiration
and chloroplast membranes of eukaryotes and in the plasma
membranes of prokaryotes. Each of the four parts of ATP synthase In the last few sections, we have looked rather closely at the
consists of a number of polypeptide subunits. key processes of cellular respiration. Now let’s take a step

174 UNIT TWO The Cell

 Inner
mitochondrial
Oxidative membrane
Citric
Pyruvate acid phosphorylation:
Glycolysis electron transport
oxidation cycle
and chemiosmosis

ATP ATP ATP

H+
H+

H+
H+
Intermembrane Protein complex Cyt c
space of electron
carriers

IV
Q
I III
ATP
synthase
Inner II
2 H+ + 1 2 O2 H2O
mitochondrial FADH2 FAD
membrane
NADH NAD+
ADP + P i ATP
(carrying electrons
from food)
H+

Mitochondrial 1 Electron transport chain 2 Chemiosmosis

matrix Electron transport and pumping of protons (H+), ATP synthesis powered by the flow
which create an H+ gradient across the membrane +
of H back across the membrane

Oxidative phosphorylation

 Figure 9.15 Chemiosmosis couples the and cytochrome c (Cyt c), move rapidly, ferrying force, a gradient of H! across the membrane.
electron transport chain to ATP electrons between the large complexes. As 2 During chemiosmosis, the protons flow back
synthesis. 1 NADH and FADH2 shuttle high- complexes I, III, and IV accept and then donate down their gradient via ATP synthase, which is
energy electrons extracted from food during electrons, they pump protons from the built into the membrane nearby. The ATP
glycolysis and the citric acid cycle into an electron mitochondrial matrix into the intermembrane synthase harnesses the proton-motive force to
transport chain built into the inner mitochondrial space. (In prokaryotes, protons are pumped phosphorylate ADP, forming ATP. Together,
membrane. The gold arrows trace the transport outside the plasma membrane.) Note that electron transport and chemiosmosis make up
of electrons, which finally pass to oxygen at the FADH2 deposits its electrons via complex II and oxidative phosphorylation.
“downhill” end of the chain, forming water. As so results in fewer protons being pumped into WHAT IF? If complex IV were nonfunctional,
Figure 9.13 showed, most of the electron the intermembrane space than occurs with could chemiosmosis produce any ATP, and if so,
carriers of the chain are grouped into four NADH. Chemical energy originally harvested how would the rate of synthesis differ?
complexes. Two mobile carriers, ubiquinone (Q) from food is transformed into a proton-motive

back and remind ourselves of its overall function: harvesting the 4 ATP produced directly by substrate-level phosphorylation
the energy of glucose for ATP synthesis. during glycolysis and the citric acid cycle to the many more mol-
During respiration, most energy flows in this sequence: ecules of ATP generated by oxidative phosphorylation. Each
glucose → NADH → electron transport chain → proton-motive NADH that transfers a pair of electrons from glucose to the elec-
force → ATP. We can do some bookkeeping to calculate the ATP tron transport chain contributes enough to the proton-motive
profit when cellular respiration oxidizes a molecule of glucose to force to generate a maximum of about 3 ATP.
six molecules of carbon dioxide. The three main departments of Why are the numbers in Figure 9.16 inexact? There are
this metabolic enterprise are glycolysis, the citric acid cycle, and three reasons we cannot state an exact number of ATP mol-
the electron transport chain, which drives oxidative phosphory- ecules generated by the breakdown of one molecule of glu-
lation. Figure 9.16, on the next page, gives a detailed accounting cose. First, phosphorylation and the redox reactions are not
of the ATP yield per glucose molecule oxidized. The tally adds directly coupled to each other, so the ratio of the number of

CHAPTER 9 Cellular Respiration and Fermentation 175

 CYTOSOL Electron shuttles MITOCHONDRION
span membrane 2 NADH
or
2 FADH2

2 NADH 2 NADH 6 NADH 2 FADH2

Glycolysis Pyruvate oxidation Oxidative

Citric phosphorylation:
Glucose 2 Pyruvate 2 Acetyl CoA acid electron transport
cycle and
chemiosmosis

+ 2 ATP + 2 ATP + about 26 or 28 ATP

by substrate-level by substrate-level by oxidative phosphorylation, depending
phosphorylation phosphorylation on which shuttle transports electrons
from NADH in cytosol

Maximum per glucose: About

30 or 32 ATP

 Figure 9.16 ATP yield per molecule of glucose at each stage of cellular respiration.
Explain exactly how the numbers “26 or 28” were calculated.
?
NADH molecules to the number of ATP molecules is not a electrons are passed to mitochondrial NAD!, as in liver cells
whole number. We know that 1 NADH results in 10 H! being and heart cells, the yield is about 2.5 ATP per NADH.
transported out across the inner mitochondrial membrane, but A third variable that reduces the yield of ATP is the use of
the exact number of H! that must reenter the mitochondrial the proton-motive force generated by the redox reactions of
matrix via ATP synthase to generate 1 ATP has long been de- respiration to drive other kinds of work. For example, the
bated. Based on experimental data, however, most biochemists proton-motive force powers the mitochondrion’s uptake of
now agree that the most accurate number is 4 H!. Therefore, a pyruvate from the cytosol. However, if all the proton-motive
single molecule of NADH generates enough proton-motive force generated by the electron transport chain were used to
force for the synthesis of 2.5 ATP. The citric acid cycle also sup- drive ATP synthesis, one glucose molecule could generate a
plies electrons to the electron transport chain via FADH2, but maximum of 28 ATP produced by oxidative phosphorylation
since its electrons enter later in the chain, each molecule of this plus 4 ATP (net) from substrate-level phosphorylation to give
electron carrier is responsible for transport of only enough H! a total yield of about 32 ATP (or only about 30 ATP if the less
for the synthesis of 1.5 ATP. These numbers also take into ac- efficient shuttle were functioning).
count the slight energetic cost of moving the ATP formed in We can now roughly estimate the efficiency of
the mitochondrion out into the cytosol, where it will be used. respiration—that is, the percentage of chemical energy in glu-
Second, the ATP yield varies slightly depending on the type cose that has been transferred to ATP. Recall that the complete
of shuttle used to transport electrons from the cytosol into oxidation of a mole of glucose releases 686 kcal of energy
the mitochondrion. The mitochondrial inner membrane is under standard conditions (∆G " 686 kcal/mol). Phosphory-
impermeable to NADH, so NADH in the cytosol is segregated lation of ADP to form ATP stores at least 7.3 kcal per mole of
from the machinery of oxidative phosphorylation. The 2 elec- ATP. Therefore, the efficiency of respiration is 7.3 kcal per mole
trons of NADH captured in glycolysis must be conveyed into of ATP times 32 moles of ATP per mole of glucose divided by
the mitochondrion by one of several electron shuttle systems. 686 kcal per mole of glucose, which equals 0.34. Thus, about
Depending on the kind of shuttle in a particular cell type, the 34% of the potential chemical energy in glucose has been
electrons are passed either to NAD! or to FAD in the mito- transferred to ATP; the actual percentage is bound to vary as
chondrial matrix (see Figure 9.16). If the electrons are passed ∆G varies under different cellular conditions. Cellular respi-
to FAD, as in brain cells, only about 1.5 ATP can result from ration is remarkably efficient in its energy conversion. By
each NADH that was originally generated in the cytosol. If the comparison, the most efficient automobile converts only

176 UNIT TWO The Cell

about 25% of the energy stored in gasoline to energy that anaerobic respiration but not in fermentation. (The electron
moves the car. transport chain is also called the respiratory chain because of
The rest of the energy stored in glucose is lost as heat. We its role in both types of cellular respiration.)
humans use some of this heat to maintain our relatively high We have already mentioned anaerobic respiration, which
body temperature (37°C), and we dissipate the rest through takes place in certain prokaryotic organisms that live in envi-
sweating and other cooling mechanisms. ronments without oxygen. These organisms have an electron
Under certain conditions, it may be beneficial to reduce the transport chain but do not use oxygen as a final electron ac-
efficiency of cellular respiration. A remarkable adaptation is ceptor at the end of the chain. Oxygen performs this function
shown by hibernating mammals, which overwinter in a state very well because it is extremely electronegative, but other,
of inactivity and lowered metabolism. Although their internal less electronegative substances can also serve as final electron
body temperature is lower than normal, it still must be kept acceptors. Some “sulfate-reducing” marine bacteria, for in-
significantly higher than the external air temperature. One stance, use the sulfate ion (SO42 ) at the end of their respira-
type of tissue, called brown fat, is made up of cells packed full tory chain. Operation of the chain builds up a proton-motive
of mitochondria. The inner mitochondrial membrane con- force used to produce ATP, but H2S (hydrogen sulfide) is pro-
tains a channel protein called the uncoupling protein, which duced as a by-product rather than water. The rotten-egg odor
allows protons to flow back down their concentration gradient you may have smelled while walking through a salt marsh or
without generating ATP. Activation of these proteins in hiber- a mudflat signals the presence of sulfate-reducing bacteria.
nating mammals results in ongoing oxidation of stored fuel Fermentation is a way of harvesting chemical energy with-
stores (fats), generating heat without any ATP production. In out using either oxygen or any electron transport chain—in
the absence of such an adaptation, the ATP level would build other words, without cellular respiration. How can food be
up to a point that cellular respiration would be shut down due oxidized without cellular respiration? Remember, oxidation
to regulatory mechanisms to be discussed later. simply refers to the loss of electrons to an electron acceptor,
so it does not need to involve oxygen. Glycolysis oxidizes
CONCEPT CHECK 9.4 glucose to two molecules of pyruvate. The oxidizing agent of
1. What effect would an absence of O2 have on the glycolysis is NAD!, and neither oxygen nor any electron
process shown in Figure 9.15? transfer chain is involved. Overall, glycolysis is exergonic,
2. WHAT IF? In the absence of O2, as in question 1, and some of the energy made available is used to produce
what do you think would happen if you decreased 2 ATP (net) by substrate-level phosphorylation. If oxygen is
the pH of the intermembrane space of the mitochon- present, then additional ATP is made by oxidative phospho-
drion? Explain your answer. rylation when NADH passes electrons removed from glucose
3. MAKE CONNECTIONS In Concept 7.1 (pp. 127–128), to the electron transport chain. But glycolysis generates
you learned that membranes must be fluid to func- 2 ATP whether oxygen is present or not—that is, whether
tion properly. How does the operation of the electron conditions are aerobic or anaerobic.
transport chain support that assertion? As an alternative to respiratory oxidation of organic nutri-
For suggested answers, see Appendix A. ents, fermentation is an extension of glycolysis that allows
continuous generation of ATP by the substrate-level phos-
phorylation of glycolysis. For this to occur, there must be a
CONCEPT
9.5 sufficient supply of NAD! to accept electrons during the oxi-
dation step of glycolysis. Without some mechanism to recy-
Fermentation and anaerobic cle NAD! from NADH, glycolysis would soon deplete the
respiration enable cells to produce cell’s pool of NAD! by reducing it all to NADH and would
shut itself down for lack of an oxidizing agent. Under aerobic
ATP without the use of oxygen conditions, NAD! is recycled from NADH by the transfer of
Because most of the ATP generated by cellular respiration is electrons to the electron transport chain. An anaerobic alter-
due to the work of oxidative phosphorylation, our estimate native is to transfer electrons from NADH to pyruvate, the
of ATP yield from aerobic respiration is contingent on an ad- end product of glycolysis.
equate supply of oxygen to the cell. Without the electronega-
tive oxygen to pull electrons down the transport chain,
Types of Fermentation
oxidative phosphorylation eventually ceases. However, there Fermentation consists of glycolysis plus reactions that regen-
are two general mechanisms by which certain cells can oxi- erate NAD! by transferring electrons from NADH to pyruvate
dize organic fuel and generate ATP without the use of oxygen: or derivatives of pyruvate. The NAD! can then be reused to
anaerobic respiration and fermentation. The distinction be- oxidize sugar by glycolysis, which nets two molecules of ATP
tween these two is that an electron transport chain is used in by substrate-level phosphorylation. There are many types of

CHAPTER 9 Cellular Respiration and Fermentation 177

fermentation, differing in the end products formed from have used yeast in brewing, winemaking, and baking. The
pyruvate. Two common types are alcohol fermentation and CO2 bubbles generated by baker’s yeast during alcohol fer-
lactic acid fermentation. mentation allow bread to rise.
In alcohol fermentation (Figure 9.17a), pyruvate is During lactic acid fermentation (Figure 9.17b), pyru-
converted to ethanol (ethyl alcohol) in two steps. The first vate is reduced directly by NADH to form lactate as an end
step releases carbon dioxide from the pyruvate, which is con- product, with no release of CO2. (Lactate is the ionized form of
verted to the two-carbon compound acetaldehyde. In the lactic acid.) Lactic acid fermentation by certain fungi and bac-
second step, acetaldehyde is reduced by NADH to ethanol. teria is used in the dairy industry to make cheese and yogurt.
This regenerates the supply of NAD! needed for the continu- Human muscle cells make ATP by lactic acid fermentation
ation of glycolysis. Many bacteria carry out alcohol fermenta- when oxygen is scarce. This occurs during strenuous exercise,
tion under anaerobic conditions. Yeast (a fungus) also carries when sugar catabolism for ATP production outpaces the
out alcohol fermentation. For thousands of years, humans muscle’s supply of oxygen from the blood. Under these con-
ditions, the cells switch from aerobic respiration to fermenta-
tion. The lactate that accumulates was previously thought to
cause muscle fatigue and pain, but recent research suggests
2 ADP + 2 P i 2 ATP O– instead that increased levels of potassium ions (K!) may be to
C O blame, while lactate appears to enhance muscle performance.
In any case, the excess lactate is gradually carried away by the
C O
Glucose Glycolysis blood to the liver, where it is converted back to pyruvate by
CH3 liver cells. Because oxygen is available, this pyruvate can then
2 Pyruvate enter the mitochondria in liver cells and complete cellular
respiration.
2 NAD+ 2 NADH 2 CO2
+ 2 H+
H H Comparing Fermentation with Anaerobic
H C OH C O
and Aerobic Respiration
CH3 CH3 Fermentation, anaerobic respiration, and aerobic respiration
are three alternative cellular pathways for producing ATP by
2 Ethanol 2 Acetaldehyde
harvesting the chemical energy of food. All three use glycoly-
(a) Alcohol fermentation sis to oxidize glucose and other organic fuels to pyruvate,
with a net production of 2 ATP by substrate-level phosphory-
lation. And in all three pathways, NAD! is the oxidizing
agent that accepts electrons from food during glycolysis.
2 ADP + 2 P i 2 ATP
A key difference among the three pathways is the contrast-
ing mechanisms for oxidizing NADH back to NAD!, which is
required to sustain glycolysis. In fermentation, the final elec-
Glucose Glycolysis O– tron acceptor is an organic molecule such as pyruvate (lactic
C O acid fermentation) or acetaldehyde (alcohol fermentation).
C O In cellular respiration, by contrast, electrons carried by
2 NAD+ 2 NADH NADH are transferred to an electron transport chain, where
O– CH3
+ 2 H+ they move stepwise down a series of redox reactions to a final
C O 2 Pyruvate electron acceptor. In aerobic respiration, the final electron ac-
H C OH ceptor is oxygen; in anaerobic respiration, the final acceptor
is another molecule that is electronegative (although invari-
CH3
ably less so than oxygen). Passage of electrons from NADH to
2 Lactate the electron transport chain not only regenerates the NAD!
(b) Lactic acid fermentation
required for glycolysis but pays an ATP bonus when the step-
wise electron transport from this NADH to oxygen drives ox-
 Figure 9.17 Fermentation. In the absence of oxygen, many cells idative phosphorylation. An even bigger ATP payoff comes
use fermentation to produce ATP by substrate-level phosphorylation. from the oxidation of pyruvate in the mitochondrion, which
Pyruvate, the end product of glycolysis, serves as an electron acceptor
is unique to respiration. Without an electron transport chain,
for oxidizing NADH back to NAD!, which can then be reused in
glycolysis. Two of the common end products formed from fermentation the energy still stored in pyruvate is unavailable to most cells.
are (a) ethanol and (b) lactate, the ionized form of lactic acid. Thus, cellular respiration harvests much more energy from

178 UNIT TWO The Cell

each sugar molecule than fermentation can. In fact, aerobic The Evolutionary Significance of Glycolysis
respiration yields up to 16 times as much ATP per glucose
molecule as does fermentation—up to 32 molecules of ATP
EVOLUTION The role of glycolysis in both fermentation
for respiration, compared with 2 molecules of ATP produced and respiration has an evolutionary basis. Ancient prokary-
by substrate-level phosphorylation in fermentation. otes are thought to have used glycolysis to make ATP long be-
Some organisms, called obligate anaerobes, carry out fore oxygen was present in Earth’s atmosphere. The oldest
only fermentation or anaerobic respiration. In fact, these or- known fossils of bacteria date back 3.5 billion years, but ap-
ganisms cannot survive in the presence of oxygen, some preciable quantities of oxygen probably did not begin to ac-
forms of which can actually be toxic if protective systems are cumulate in the atmosphere until about 2.7 billion years ago.
not present in the cell. A few cell types, such as cells of the Cyanobacteria produced this O2 as a by-product of photosyn-
vertebrate brain, can carry out only aerobic oxidation of thesis. Therefore, early prokaryotes may have generated ATP
pyruvate, not fermentation. Other organisms, including exclusively from glycolysis. The fact that glycolysis is today
yeasts and many bacteria, can make enough ATP to survive the most widespread metabolic pathway among Earth’s or-
using either fermentation or respiration. Such species are ganisms suggests that it evolved very early in the history of
called facultative anaerobes. On the cellular level, our life. The cytosolic location of glycolysis also implies great an-
muscle cells behave as facultative anaerobes. In such cells, tiquity; the pathway does not require any of the membrane-
pyruvate is a fork in the metabolic road that leads to two al- bounded organelles of the eukaryotic cell, which evolved
ternative catabolic routes (Figure 9.18). Under aerobic condi- approximately 1 billion years after the prokaryotic cell. Gly-
tions, pyruvate can be converted to acetyl CoA, and colysis is a metabolic heirloom from early cells that contin-
oxidation continues in the citric acid cycle via aerobic respi- ues to function in fermentation and as the first stage in the
ration. Under anaerobic conditions, lactic acid fermentation breakdown of organic molecules by respiration.
occurs: Pyruvate is diverted from the citric acid cycle, serving CONCEPT CHECK 9.5
instead as an electron acceptor to recycle NAD!. To make the
1. Consider the NADH formed during glycolysis. What
same amount of ATP, a facultative anaerobe has to consume
is the final acceptor for its electrons during fermenta-
sugar at a much faster rate when fermenting than when
tion? What is the final acceptor for its electrons dur-
respiring.
ing aerobic respiration?
2. WHAT IF? A glucose-fed yeast cell is moved from an
aerobic environment to an anaerobic one. How
Glucose would its rate of glucose consumption change if ATP
were to be generated at the same rate?
Glycolysis
CYTOSOL For suggested answers, see Appendix A.

Pyruvate
No O2 present:
Fermentation
O2 present:
Aerobic cellular
CONCEPT
9.6
respiration
Glycolysis and the citric acid
cycle connect to many other
MITOCHONDRION metabolic pathways
Ethanol, Acetyl CoA
lactate, or So far, we have treated the oxidative breakdown of glucose in
other products
Citric isolation from the cell’s overall metabolic economy. In this
acid section, you will learn that glycolysis and the citric acid cycle
cycle
are major intersections of the cell’s catabolic and anabolic
(biosynthetic) pathways.

The Versatility of Catabolism

 Figure 9.18 Pyruvate as a key juncture in catabolism.
Glycolysis is common to fermentation and cellular respiration. The end Throughout this chapter, we have used glucose as the fuel for
product of glycolysis, pyruvate, represents a fork in the catabolic cellular respiration. But free glucose molecules are not com-
pathways of glucose oxidation. In a facultative anaerobe or a muscle
mon in the diets of humans and other animals. We obtain
cell, which are capable of both aerobic cellular respiration and
fermentation, pyruvate is committed to one of those two pathways, most of our calories in the form of fats, proteins, sucrose and
usually depending on whether or not oxygen is present. other disaccharides, and starch, a polysaccharide. All these

CHAPTER 9 Cellular Respiration and Fermentation 179

 Proteins Carbohydrates Fats acids can feed into glycolysis or the citric acid cycle, their
amino groups must be removed, a process called deamination.
The nitrogenous refuse is excreted from the animal in the
Amino Sugars Glycerol Fatty form of ammonia (NH3), urea, or other waste products.
acids acids Catabolism can also harvest energy stored in fats obtained
either from food or from storage cells in the body. After fats
are digested to glycerol and fatty acids, the glycerol is
Glycolysis converted to glyceraldehyde 3-phosphate, an intermediate of
Glucose glycolysis. Most of the energy of a fat is stored in the fatty
acids. A metabolic sequence called beta oxidation breaks
the fatty acids down to two-carbon fragments, which enter
Glyceraldehyde 3- P the citric acid cycle as acetyl CoA. NADH and FADH2 are also
generated during beta oxidation; they can enter the electron
NH3 transport chain, leading to further ATP production. Fats
Pyruvate
make excellent fuel, in large part due to their chemical struc-
ture and the high energy level of their electrons (equally
shared between carbon and hydrogen) compared to those of
Acetyl CoA
carbohydrates. A gram of fat oxidized by respiration produces
more than twice as much ATP as a gram of carbohydrate. Un-
fortunately, this also means that a person trying to lose
weight must work hard to use up fat stored in the body be-
Citric
cause so many calories are stockpiled in each gram of fat.
acid
cycle

Biosynthesis (Anabolic Pathways)

Cells need substance as well as energy. Not all the organic
molecules of food are destined to be oxidized as fuel to make
Oxidative ATP. In addition to calories, food must also provide the car-
phosphorylation
bon skeletons that cells require to make their own mol-
 Figure 9.19 The catabolism of various molecules from ecules. Some organic monomers obtained from digestion
food. Carbohydrates, fats, and proteins can all be used as fuel for can be used directly. For example, as previously mentioned,
cellular respiration. Monomers of these molecules enter glycolysis or
amino acids from the hydrolysis of proteins in food can be
the citric acid cycle at various points. Glycolysis and the citric acid cycle
are catabolic funnels through which electrons from all kinds of organic incorporated into the organism’s own proteins. Often, how-
molecules flow on their exergonic fall to oxygen. ever, the body needs specific molecules that are not present
as such in food. Compounds formed as intermediates of gly-
colysis and the citric acid cycle can be diverted into anabolic
pathways as precursors from which the cell can synthesize
organic molecules in food can be used by cellular respiration the molecules it requires. For example, humans can make
to make ATP (Figure 9.19). about half of the 20 amino acids in proteins by modifying
Glycolysis can accept a wide range of carbohydrates for ca- compounds siphoned away from the citric acid cycle; the
tabolism. In the digestive tract, starch is hydrolyzed to glu- rest are “essential amino acids” that must be obtained in the
cose, which can then be broken down in the cells by diet. Also, glucose can be made from pyruvate, and fatty
glycolysis and the citric acid cycle. Similarly, glycogen, the acids can be synthesized from acetyl CoA. Of course, these
polysaccharide that humans and many other animals store anabolic, or biosynthetic, pathways do not generate ATP, but
in their liver and muscle cells, can be hydrolyzed to glucose instead consume it.
between meals as fuel for respiration. The digestion of disac- In addition, glycolysis and the citric acid cycle function as
charides, including sucrose, provides glucose and other metabolic interchanges that enable our cells to convert some
monosaccharides as fuel for respiration. kinds of molecules to others as we need them. For example,
Proteins can also be used for fuel, but first they must be di- an intermediate compound generated during glycolysis, di-
gested to their constituent amino acids. Many of the amino hydroxyacetone phosphate (see Figure 9.9, step 5), can be
acids are used by the organism to build new proteins. Amino converted to one of the major precursors of fats. If we eat
acids present in excess are converted by enzymes to interme- more food than we need, we store fat even if our diet is fat-
diates of glycolysis and the citric acid cycle. Before amino free. Metabolism is remarkably versatile and adaptable.

180 UNIT TWO The Cell

 Glucose

Regulation of Cellular Respiration AMP

Glycolysis
via Feedback Mechanisms Fructose 6-phosphate Stimulates
+
Basic principles of supply and demand regulate the metabolic Phosphofructokinase
economy. The cell does not waste energy making more of a par- –
–
ticular substance than it needs. If there is a glut of a certain Fructose 1,6-bisphosphate
amino acid, for example, the anabolic pathway that synthe- Inhibits Inhibits
sizes that amino acid from an intermediate of the citric acid
cycle is switched off. The most common mechanism for this
control is feedback inhibition: The end product of the anabolic
pathway inhibits the enzyme that catalyzes an early step of the
pathway (see Figure 8.21). This prevents the needless diversion
of key metabolic intermediates from uses that are more urgent.
Pyruvate
The cell also controls its catabolism. If the cell is working
hard and its ATP concentration begins to drop, respiration
ATP Citrate
speeds up. When there is plenty of ATP to meet demand, respi- Acetyl CoA
ration slows down, sparing valuable organic molecules for
other functions. Again, control is based mainly on regulating
the activity of enzymes at strategic points in the catabolic
pathway. As shown in Figure 9.20, one important switch is
Citric
phosphofructokinase, the enzyme that catalyzes step 3 of gly- acid
colysis (see Figure 9.9). That is the first step that commits the cycle
substrate irreversibly to the glycolytic pathway. By controlling
the rate of this step, the cell can speed up or slow down the en-
tire catabolic process. Phosphofructokinase can thus be con-
sidered the pacemaker of respiration. Oxidative
Phosphofructokinase is an allosteric enzyme with receptor phosphorylation
sites for specific inhibitors and activators. It is inhibited by ATP
 Figure 9.20 The control of cellular respiration. Allosteric
and stimulated by AMP (adenosine monophosphate), which enzymes at certain points in the respiratory pathway respond to inhibitors
the cell derives from ADP. As ATP accumulates, inhibition of the and activators that help set the pace of glycolysis and the citric acid cycle.
enzyme slows down glycolysis. The enzyme becomes active Phosphofructokinase, which catalyzes an early step in glycolysis (see
Figure 9.9), is one such enzyme. It is stimulated by AMP (derived from
again as cellular work converts ATP to ADP (and AMP) faster
ADP) but is inhibited by ATP and by citrate. This feedback regulation adjusts
than ATP is being regenerated. Phosphofructokinase is also sen- the rate of respiration as the cell’s catabolic and anabolic demands change.
sitive to citrate, the first product of the citric acid cycle. If citrate
accumulates in mitochondria, some of it passes into the cytosol
CONCEPT CHECK 9.6
and inhibits phosphofructokinase. This mechanism helps syn- 1. MAKE CONNECTIONS Compare the structure of a fat
chronize the rates of glycolysis and the citric acid cycle. As cit- (see Figure 5.10, p. 75) with that of a carbohydrate
rate accumulates, glycolysis slows down, and the supply of (see Figure 5.3, p. 70). What features of their struc-
acetyl groups to the citric acid cycle decreases. If citrate con- tures make fat a much better fuel?
sumption increases, either because of a demand for more ATP or 2. Under what circumstances might your body synthe-
because anabolic pathways are draining off intermediates of the size fat molecules?
citric acid cycle, glycolysis accelerates and meets the demand. 3. MAKE CONNECTIONS Return to Figure 5.6b on
Metabolic balance is augmented by the control of enzymes that page 72 and look at the arrangement of glycogen
catalyze other key steps of glycolysis and the citric acid cycle. and mitochondria in the micrograph. What is the
Cells are thrifty, expedient, and responsive in their metabolism. connection between glycogen and mitochondria?
Cellular respiration and metabolic pathways play a role of 4. WHAT IF? What will happen in a muscle cell that
central importance in organisms. Examine Figure 9.2 again to has used up its supply of oxygen and ATP? (Review
put cellular respiration into the broader context of energy flow Figures 9.18 and 9.20.)
and chemical cycling in ecosystems. The energy that keeps us 5. WHAT IF? During intense exercise, can a muscle cell
alive is released, not produced, by cellular respiration. We are use fat as a concentrated source of chemical energy?
tapping energy that was stored in food by photosynthesis. In Explain. (Review Figures 9.18 and 9.19.)
the next chapter, you will learn how photosynthesis captures For suggested answers, see Appendix A.
light and converts it to chemical energy.

CHAPTER 9 Cellular Respiration and Fermentation 181

 9 CHAPTER REVIEW
SUMMARY OF KEY CONCEPTS CONCEPT 9.4
During oxidative phosphorylation, chemiosmosis couples
CONCEPT 9.1 electron transport to ATP synthesis (pp. 172–177)
Catabolic pathways yield energy by oxidizing organic fuels • NADH and FADH2 transfer electrons to the electron transport
(pp. 164–168) chain. Electrons move down the chain, losing energy in several
energy-releasing steps. Finally, electrons are passed to O2, re-
• Cells break down glucose and other organic fuels to yield chemi-
ducing it to H2O.
cal energy in the form of ATP. Fermentation is a partial degra-
dation of glucose without the use of oxygen. Cellular INTERMEMBRANE
respiration is a more complete breakdown of glucose; in SPACE H+
H+
aerobic respiration, oxygen is used as a reactant. The cell taps
the energy stored in food molecules through redox reactions, H+
in which one substance partially or totally shifts electrons to Protein complex Cyt c
of electron
another. Oxidation is the loss of electrons from one substance, carriers
while reduction is the addition of electrons to the other.
• During aerobic respiration, glucose (C6H12O6) is oxidized to CO2, IV
Q
and O2 is reduced to H2O. Electrons lose potential energy during III
I
their transfer from glucose or other organic compounds to oxy-
gen. Electrons are usually passed first to NAD , reducing it to II
2 H+ + 1 2 O2 H2O
NADH, and then from NADH to an electron transport FADH2 FAD
chain, which conducts them to O2 in energy-releasing steps.
NADH NAD+
The energy is used to make ATP. MITOCHONDRIAL MATRIX
(carrying electrons from food)
• Aerobic respiration occurs in three stages: (1) glycolysis, (2) pyru-
vate oxidation and the citric acid cycle, and (3) oxidative
• At certain steps along the INTER-
phosphorylation (electron transport and chemiosmosis). MEMBRANE
electron transport chain, SPACE H+
Describe the difference between the two processes in cellular electron transfer causes pro-
? respiration that produce ATP: oxidative phosphorylation and tein complexes to move H!
substrate-level phosphorylation. from the mitochondrial ma-
trix (in eukaryotes) to the in-
CONCEPT 9.2 termembrane space, storing
Glycolysis harvests chemical energy by oxidizing glucose to energy as a proton-motive
MITO- ATP
pyruvate (pp. 168–169) force (H! gradient). As H! CHONDRIAL synthase
diffuses back into the matrix MATRIX
Inputs Outputs through ATP synthase, its
Glycolysis passage drives the phospho-
Glucose 2 Pyruvate + 2 ATP + 2 NADH rylation of ADP, a process ADP + P i H+ ATP
called chemiosmosis.
• About 34% of the energy stored in a glucose molecule is trans-
What is the source of energy for the formation of ATP and ferred to ATP during cellular respiration, producing a maximum
? NADH in glycolysis? of about 32 ATP.
Briefly explain the mechanism by which ATP synthase produces
CONCEPT 9.3 ? ATP. List three locations in which ATP synthases are found.
After pyruvate is oxidized, the citric acid cycle completes the
energy-yielding oxidation of organic molecules (pp. 170–172) CONCEPT 9.5
• In eukaryotic cells, pyruvate enters the mitochondrion and is Fermentation and anaerobic respiration enable cells to
oxidized to acetyl CoA, which is further oxidized in the citric produce ATP without the use of oxygen (pp. 177–179)
acid cycle. • Glycolysis nets 2 ATP by substrate-level phosphorylation,
whether oxygen is present or not. Under anaerobic conditions,
Inputs Outputs
either anaerobic respiration or fermentation can take place. In
anaerobic respiration, an electron transport chain is present
2 Pyruvate 2 Acetyl CoA 2 ATP 8 NADH with a final electron acceptor other than oxygen. In fermenta-
Citric tion, the electrons from NADH are passed to pyruvate or a de-
2 Oxaloacetate acid rivative of pyruvate, regenerating the NAD! required to oxidize
cycle 6 CO2 2 FADH 2 more glucose. Two common types of fermentation are alcohol
fermentation and lactic acid fermentation.
• Fermentation and anaerobic or aerobic respiration all use gly-
colysis to oxidize glucose, but they differ in their final electron
What molecular products indicate the complete oxidation of
? glucose during cellular respiration?
acceptor and whether an electron transport chain is used (respi-
ration) or not (fermentation). Respiration yields more ATP;

182 UNIT TWO The Cell

 aerobic respiration, with O2 as the final electron acceptor, yields c. The electrons gain free energy.
about 16 times as much ATP as does fermentation. d. The cytochromes phosphorylate ADP to form ATP.
• Glycolysis occurs in nearly all organisms and is thought to have e. NAD! is oxidized.
evolved in ancient prokaryotes before there was O2 in the 7. Most CO2 from catabolism is released during
atmosphere. a. glycolysis. d. electron transport.
Which process yields more ATP, fermentation or anaerobic respi- b. the citric acid cycle. e. oxidative phosphorylation.
? ration? Explain. c. lactate fermentation.

CONCEPT 9.6 LEVEL 3: SYNTHESIS/EVALUATION

8. DRAW IT The graph here shows
Glycolysis and the citric acid cycle connect to many other
metabolic pathways (pp. 179–181) the pH difference across the inner

across membrane
mitochondrial membrane over time

pH difference
• Catabolic pathways funnel electrons from many kinds of or- in an actively respiring cell. At the
ganic molecules into cellular respiration. Many carbohydrates time indicated by the vertical arrow,
can enter glycolysis, most often after conversion to glucose. a metabolic poison is added that
Amino acids of proteins must be deaminated before being oxi- specifically and completely inhibits
dized. The fatty acids of fats undergo beta oxidation to two- all function of mitochondrial ATP Time
carbon fragments and then enter the citric acid cycle as acetyl synthase. Draw what you would ex-
CoA. Anabolic pathways can use small molecules from food di- pect to see for the rest of the graphed line.
rectly or build other substances using intermediates of glycoly-
sis or the citric acid cycle. 9. EVOLUTION CONNECTION
• Cellular respiration is controlled by allosteric enzymes at key ATP synthases are found in the prokaryotic plasma membrane
points in glycolysis and the citric acid cycle. and in mitochondria and chloroplasts. What does this suggest
about the evolutionary relationship of these eukaryotic or-
Describe how the catabolic pathways of glycolysis and the citric acid ganelles to prokaryotes? How might the amino acid sequences
? cycle intersect with anabolic pathways in the metabolism of a cell. of the ATP synthases from the different sources support or re-
fute your hypothesis?
TEST YOUR UNDERSTANDING 10. SCIENTIFIC INQUIRY
In the 1930s, some physicians prescribed low doses of a com-
LEVEL 1: KNOWLEDGE/COMPREHENSION pound called dinitrophenol (DNP) to help patients lose weight.
This unsafe method was abandoned after some patients died.
1. The immediate energy source that drives ATP synthesis by ATP
DNP uncouples the chemiosmotic machinery by making the
synthase during oxidative phosphorylation is the
lipid bilayer of the inner mitochondrial membrane leaky to H!.
a. oxidation of glucose and other organic compounds.
Explain how this could cause weight loss and death.
b. flow of electrons down the electron transport chain.
c. affinity of oxygen for electrons. 11. WRITE ABOUT A THEME
d. H! concentration across the membrane holding ATP synthase. Emergent Properties In a short essay (100–150 words),
e. transfer of phosphate to ADP. explain how oxidative phosphorylation—the production
of ATP using energy derived from the redox reactions of a
2. Which metabolic pathway is common to both fermentation
spatially organized electron transport chain followed by
and cellular respiration of a glucose molecule?
chemiosmosis—is an example of how new properties
a. the citric acid cycle
emerge at each level of the biological hierarchy.
b. the electron transport chain
c. glycolysis For selected answers, see Appendix A.
d. synthesis of acetyl CoA from pyruvate
e. reduction of pyruvate to lactate
3. In mitochondria, exergonic redox reactions www.masteringbiology.com
a. are the source of energy driving prokaryotic ATP synthesis.
b. are directly coupled to substrate-level phosphorylation. 1. MasteringBiology® Assignments
c. provide the energy that establishes the proton gradient. Tutorials Cellular Respiration: Inputs and Outputs •
d. reduce carbon atoms to carbon dioxide. Glycolysis • Acetyl CoA Formation and the Citric Acid Cycle •
e. are coupled via phosphorylated intermediates to ender- Oxidative Phosphorylation • Summary
gonic processes. Tutorial Pathways for Pyruvate
Activities Build a Chemical Cycling System • Overview of Cellular
4. The final electron acceptor of the electron transport chain
Respiration • Redox Reactions • Glycolysis • The Citric Acid Cycle •
that functions in aerobic oxidative phosphorylation is
Electron Transport • Fermentation • Glucose Metabolism •
a. oxygen. b. water. c. NAD!. d. pyruvate. e. ADP. Discovery Channel Video: Space Plants
Questions Student Misconceptions • Reading Quiz • Multiple
LEVEL 2: APPLICATION/ANALYSIS
Choice • End-of-Chapter
5. What is the oxidizing agent in the following reaction?
2. eText
Pyruvate ! NADH ! H! S Lactate ! NAD! Read your book online, search, take notes, highlight text, and more.
a. oxygen b. NADH c. NAD! d. lactate e. pyruvate
3. The Study Area
6. When electrons flow along the electron transport chains of Practice Tests • Cumulative Test • 3-D Animations •
mitochondria, which of the following changes occurs? MP3 Tutor Sessions • Videos • Activities • Investigations • Lab
a. The pH of the matrix increases. Media • Audio Glossary • Word Study Tools • Art
b. ATP synthase pumps protons by active transport.

CHAPTER 9 Cellular Respiration and Fermentation 183