The problem of alcoholic fermentation,of the origin and natureof

change,which converted
and apparentlyspontaneous
that mysterious
g i n e , s e e m st o h a v ee x t h e i n s i p i dj u i c eo f t h e g r a p ei n t o s t i m u l a t i n w
erted a fascinationover the minds of naturalphilosophersfrom the
very earliesttimes.
-Arthur Harden,Alcoholic Fermentation,
7923

and
GIuconeogenesis,
Glycolysis,
Pathway
Phosphate
thePentose
528
14.1 Glycolysis
forGlycolysis543
Pathways
14.2 Feeder
Anaerobic
Conditions:
ofPyruvate
under
14.3 Fates
546
Fermentation
551
14.4 Gluconeogenesis
Oxidation558
Pathway
ofGlucose
Phosphate
14.5 Pentose
lucose occupiesa central position in the metabolism of plants,animals,and many microorganisms.
It is relatively rich in potential energy,and thus a
good fuel; the completeoxidationof glucoseto carbon
dioxide and water proceedswith a standardfree-energy
changeof -2,840 kJ/mol.By storingglucoseas a high
molecularweight pol;rmersuchas starchor glycogen,a
cell can stockpile large quantities of hexoseunits while
maintaining a relatively low c;'tosolic osmolarity.When
energydemandsincrease,glucosecanbe releasedfrom
these intracellularstoragepol)rmersand used to produce ATP either aerobicallyor anaerobically.
Glucoseis not only an excellent fuel, it is also a remarkably versatile precursor, capableof supplying a
huge array of metabolic intermediatesfor biosl'nthetic
coLi,can obreactions.A bacteriumsuchas,Uscheri'chict'
glucose
the carbon skeletonsfor every amino
tain from
acid, nucleotide,coen4.'rne,fatty acid, or other metabolic intermediateit needsfor growth. A comprehensive
studyof the metabolicfatesof glucosewould encompass
hundredsor thousandsof transformations.In animals
and vascularplants,glucosehas four major fates:it may
desbe usedin the synthesisof complexpolysaccharides
tined for the extracellularspace;stored in cells (as a
polysaccharide
or assucrose);oxidizedto a three-carbon
compound (pyruvate) via glycolysisto provide ATP and

metabolicintermediates;or oxidized via the pentose

phosphate(phosphogluconate)pathway to yield ribose
5-phosphatefor nucleic acid synthesisand NADPH for
reductivebiosyntheticprocesses(FiS. 14-1 ).
Organismsthat do not have accessto glucosefrom
other sourcesmust make it. Photosyntheticorganisms
make glucose by first reducing atmosphericCO2 to
trioses,then convertingthe triosesto glucose.Nonphotosyntheticcellsmakeglucosefrom simplerthree- and
four-carbonprecursorsby the processofgluconeogenesis, effectively reversing glycolysis in a pathway that
usesmany of the glycolytic enzyrnes.
In this chapter we describethe individual reactions
gluconeogenesis,
and the pentosephosglycolysis,
of
phate pathway and the functional signiflcanceof each
pathway.We alsodescribethe variousmetabolicfates of
the pyruvateproducedby glycolysis.They include the
fermentations that are used by many organismsin
anaerobicnichesto produceATP and that are exploited
industrialtyas sourcesof ethanol,lactic acid,and other
Extracellular matrix
and cell wall
polysaccharides

GlYcogen,
starch, sucrose

\/

synthesisof

oxidationvia

s'iorace

;:il*:13'

ilil$;"on*on^7
/
Ribose 5-phosphate
14-1 Major pathwaysof glucoseutilization.Althoughnot the
FIGURE
only possiblefatesfor glucose,thesefour pathwaysarethe mostsignificantin termsof theamountofglucosethatflowsthroughthemin most
cells.

Grr)

F"l

G l y c o l yG
s i lsu, c o n e o g e naensdtihse, p e n t o speh o s p h aptaet h w a y

commerciallyusefulproducts.And we look at the path_

waysthat feed varioussugarsfrom mono-, di-, and polysaccharidesinto the glycol;,tic pathway.The discussion
of glucosemetabolismcontinuesin Chapter 1b, where
we use the processesof carbohydratesynthesisand
degradationto illustrate the manymechanismsby which
organismsregulate metabolic pathways.The biosynthetic pathwaysfrom glucoseto extracellularmatrix
and cell wall polysaccharides
and storagepolysaccharides are discussedin Chapter20.

14JlGlycolysis
In glycolysis (from the Greek glykys, ,,sweet" or
"sugar,"andlgsi,s,"splitting"),a moleculeof glucoseis
degradedin a series of enzl'rne-catalyzedreactions to
yield two molecules of the three-carbon compound
pyruvate. During the sequentialreactions of glycolysis,
some of the free energyreleasedfrom glucoseis conservedin the form of ATP and NADH Glycolysiswas the
first metabolicpathwayto be elucidatedand is probably
the best understood.From EduardBuchner'sdiscovery
in 1897of fermentationin brokenextractsof yeastcells
until the elucidation of the whole pathway in yeast (by
Otto Warburgand Hansvon Euler-Chelpin)and in muscle (by Gustav Embden and Otto Meyerhof) in the
1930s,the reactionsofglycolysisin extractsofyeastand
musclewere a majorfocusof biochemicalresearch.The
philosophicalshift that accompaniedthese discoveries
wasannouncedby JacquesLoebin 1906:
Through the discovery of Buchner, Biology was
relieved of another fragment of mysticism. The
splitting up of sugarinto CO2and alcoholis no more
the effect of a "vital principle" than the splitting up
of canesugarby invertase.The history of this problem is instructive, as it warns us againstconsidering
problemsas beyond our reach becausethey have
not yet found their solution.
The development of methods of enz1nnepurif,ca_
tion, the discoveryand recognitionof the importanceof
coenzyrressuch as NAD, and the discoveryof the piv_
otal metabolicrole of ATP and other phosphorylated

Hansvon Euler-Chelpin,
1873-1964

CustavEmbden,
1874-1933

compoundsall cameout of studiesof glycolysis.The glycolytic enzyrnesof many specieshave long since been
purifled and thoroughly studied.
Glycolysisis an almost universalcentral pathway of
glucosecatabolism,the pathwaywith the largestflux of
carbon in most cells. The glycoly'ticbreakdown of glucose is the sole source of metabolic energy in some
mammaliantissues and cell types (erythrocytes, renal
medulla,brain, and sperm, for example).Someplant
tissuesthat are modifledto storestarch (suchaspotato
tubers) and some aquatic plants (watercress, for
example) derive most of their energy from glycolysis;
many anaerobicmicroorganismsare entirely dependent
on glycolysis.
Fermentation is a generalterm for the anaerobi,c
degradationof glucoseor other organicnutrients to obtain energy,conservedasATP.Becauseliving organisms
first arosein an atmospherewithout oxygen,anaerobic
breakdownof glucoseis probably the most ancient biological mechanism for obtaining energy from organic
fuel molecules.And asgenomesequencingof a wide variety of orgarLismshas revealed,some archaeaand some
parasiticmicroorganisms
lack one or more of the enzyrnes
of glycolysisbut retain the core of the pathway;they presumablycarry out variant forms of glycolysis.In the corrse
of evolution,the chemistryof this reactionsequencehas
beencompletelyconserued;
the $yco$tic enz;rmesof vertebrates are closely similal, in amino acid sequenceand
three-dimensionalstructure, to their homologsin yeast
and spinach.Glycolysisdjffers amongspeciesonly in the
detailsof its regulationand in the subsequentmetabolic
fate of the pyruvate formed. The thermodlnamic principles and the types of regulatory mechanismsthat govern
$ycolysis are corrmon to all pathwaysof cell metabolism.
The glycolytic pathway,of central imporfancein itseJf,can
alsoserveasa modelfor manyaspectsof the pathwaysdiscussedthroughoutthis book.
Before examiningeachstep of the pathwayin some
detail, we take a look at glycolysisas a whole.

AnOverview:Glycolysis
Has
TwoPhases

The breakdownof the six-carbonglucoseinto two molecules of the three-carbonpyruvate occurs in l0 steps, the first 5 of which
constitute the preparatorg phase
(Fig. l4-2a). In thesereactions,gucoseis first phosphorylatedat the hydroxyl group on C-6 (step @;. fire
o-$ucose 6-phosphatethus formed is
converted to l-fructose 6-phosphate
(step @), which is again phosphorylated, this time at C-1, to fleld n-fructose 1,6-bisphosphate
(step @). For
both phosphorylations,
ATPis the phosphoryl group donor.As all sugarderivaOtto Meyerhof,
1884-.1
951
tives in glycolysisare the D isomers,we

is
1 4 . 1G l y c o l y s[rrr]
6

(a)

Preparatory

Glucose

phase

Phosphorylation of glucose
and its conversion to
glyceraldehyde 3-phosphate

first
primins [)
reaclon

HOH

Glucose 6-phosphate

1l
ll

rAr ll
\?

ll

/6\

tI tl

\7

Hexokinase
Phosphohexose
lSOmerase

Fructose 6-phosphate
Phospho@ 1l'uctokinase-1

second
priming
reaction

(3J

cHr-O-@

Fructose 1,6-bisphosphate

Glyceraldehyde 3-phosphate
+
Dihydroxyacetone phosphate
a-\
!D'

@ Aldolase
/}'
,

Trrose
phosphate
lsomerase

//o
^
eFo-cH,-9H-c\
OHH

@-o-cur-c-cH,oH
o
Payoff phase

Glyceraldehyde 3-phosphate (2)

(FFo-cs,-c

t-""o'H

3"

@ Glyceraldehyde
3-phosphate

1,3-Bisphosphoglycerate(2)

1lr 2

firstArF-

forming reaction 61 lI
v
(substratelwel
ll\
phosphorylatioo)
ll,

dehydrogenase

-q

Phosphoglycerate
kinase

3-Phosphoglyc er ate (2)

1l

@il
It

2-Phosphoglycerate(2)

1t

O
- [ lb zn,o
Phosphoenolpyruvate (2)
secondATPf:*p-eI:"fi.'"

lf
6b) [,
(subgfiare-revel
l\

phosphoqdf,tion)

"

CH"-CH-C(

J" J

'o-

I
(ry

Phosphoglycerate
mutase
@ Enolase

,o
cH,:f-c\
do-

I
(ry

Pyruvate

Oxidative conversion of
glyceraldehyde 3-phosphate to
pymvate and the coupled
formation ofATP and NADH

(2)

//o
cH3-c-c\
do-

FIGURE
14-2 The two phasesof glycolysis.Foreach moleculeof gluphase(a), two moleculesof
cosethat passesthroughthe preparatory
3-phosphate
are formed;both passthroughthe payoff
glyceraldehyde
(b).
is
end
productof the secondphaseof glycolysis.
phase Pyruvate the
ATPare consumedin the preparatory
molecule,
hvo
glucose
Foreach
phaseand fourATPareproducedin the payoffphase,givinga netyield

of two ATPper moleculeof glucoseconvertedto pyruvate'The numberedreactionstepsare catalyzedby the enzymeslistedon the right,
to the numberedheadingsin the text discussion'
and alsocorrespond
hereat @ ha'
Keepin mind that eachphosphorylgroup,represented
(-PO3_)'
two negativecharges

f-I

s i lsu, c o n e o g e naensdtihs e, p e n t o speh o s p h aptaet h w a y

i 5 3 0 _ l G l y c o l yG
will usually omit the n designation except when emphasizing stereochemistry.
Fructose 1,6-bisphosphate is split to yield two
three-carbon molecules, dihydroxyacetone phosphate
and glyceraldehyde 3-phosphate (step @); this is the ,,ly_
sis" step that gives the pathway its name. The dihydroxyacetone phosphate is isomerized to a second molecule of
glyceraldehyde 3-phosphate (step
@), ending the first
phase of glycolysis. From a chemical perspective, the isomerization rn step @ is critical for setting up the phosphorylation and C-C bond cleavage reactions in steps
@ and @, ur detailed later. Note that two molecules of
ATP are invested before the cleavage of $ucose into two
three-carbon pieces; there wrll be a good return on this
investment To summarize: in the preparatory phase of
glycolysis the energy of ATP is invested, raising the freeenergy content of the intermediates, and the carbon
chains of all the metabolized hexoses are converted to a
conunon product, glyceraldehyde 3-phosphate.
The energy gain comes rn the pagolf phase of glycolysis (Fig. 14-2b). Each molecule of glyceraldehyde
3-phosphate is oxidized and phosphorylated by inorganic phosphate (not by ATp) to form 1,3-bisphospho_
glycerate (step @). Energy is then released as the two
molecules of 1,3-bisphosphoglycerateare converted to
two molecules of pyruvate (steps @ through
@;. nluctr
of this energy is conserved by the coupled phoiphorylation of four molecules of ADp to ATp The net yield is
two molecules of ATP per molecule of glucose used, be_
cause two molecules of ATp were invested in the
preparatory phase. trnergy is also conserved in the pay_
off phase in the formation of two molecules of the electron carrier NADH per molecule of glucose.
In the sequential reactions ofglycolysis, three types
of chemical transformations are particularly noteworthy: (1) degradation ofthe carbon skeleton ofglucose to
yield pyruvate; (2) phosphorylation of ADp to ATp by
compounds with high phosphoryl group transfer poten_
tial, formed during glycolysis; and (B) transfer of a hy_
dride ion to NAD*, forming NADH.
Fates of Pguvate
With the exception of some interesting variations in the bacterial realm, the pyruvate
formed by glycolysis is further metabolized via one of
three catabolic routes. In aerobicorganisms or tissues,
under aerobic conditions, glycolysis is only the first stage
in the complete degradation of glucose (Fig. f 4-3)
Pyruvate is oxidized, with loss of its carboxyl group as
CO2,to yield the acetyl group of acetyl-coenz).rneA; the
acetyl group is then oxidized completely to CO2 by the
citric acid cycle (Chapter 16). The electrons from these
oxidations are passed to 02 through a chain of carriers
in mitochondria, to form H2O. The energy from the
electron-transfer reactions drives the slmthesis of ATp in
mitochondria (Chapter t9).
The second route for pyruvate is its reduction to
lactate via lactic acid fermentation.
When vigorously

2 Ethanol + 2CO2
Fermentation to ethanol
in yeast

Animal, plant, and

many microbial cells
under aerobic conditions
FIGURE
14-3 Threepossiblecatabolicfatesofthe pyruvateformedin
glycolysis.Pyruvatealsoservesas a precursorin manyanabolicreactions,not shownhere.

contractingskeletalmuscle must function under lowoxygen conditions (hypoxia), NADH cannot be reoxidized to NAD+, but NAD+ is required as an electron
acceptor for the further oxidation of pyruvate. Under
theseconditionspytuvateis reducedto lactate,accepting electronsfrom NADH and thereby regeneratingthe
NAD+ necessaryfor glycolysisto continue.Certaintissuesand cell types (retina and erythrocytes,for example) convert glucose to lactate even under aerobic
conditions,and lactate is also the product of glycolysis
under anaerobicconditions in some microorganisms
(Fis. 14-3).
The third major route of pyruvate catabolismleads
to ethanol.In someplant tissuesand in certain invertebrates,protists,and microorganisms
suchasbrewer'sor
baker's yeast, pyruvate is converted under hypoxic or
anaerobicconditions to ethanol and CO2,a process
called ethanol (alcohol) fermentation (Fig. 14-g).
The oxidation of pyruvate is an important catabolic
process,but pyruvatehas anabolicfatesas well. It can,
for example,providethe carbonskeletonfor the synthesis of the amino acid alanineor for the syrrthesisof fatty
acids.We return to these anabolicreactionsof pyruvate
in later chapters.
ATP and NADH Formation Coupled to Glycolysis
During glycolysissomeof the energyof the glucosemolecule is conservedin ATP, while much remains in the
product, pyruvate. The overall equationfor glycolysisis
Glucose + 2NAD* + 2ADp -f 2pi--->
2 pyruvate + 2NADH + 2H+ + 2ATp + 2H2O

(14-1)