Prescott−Harley−Klein: III. Microbial Metabolism 9.

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9.2 The Breakdown of Glucose to Pyruvate 177

from a single glucose (one by way of dihydroxyacetone phosphate),

Glucose the three-carbon stage generates four ATPs and two NADHs per
ATP
glucose. Subtraction of the ATP used in the six-carbon stage from
ADP
that produced in the three-carbon stage gives a net yield of two ATPs
per glucose. Thus the catabolism of glucose to pyruvate in glycoly-
Glucose 6-phosphate
sis can be represented by the following simple equation.
Six-carbon stage
Glucose 2ADP 2Pi 2NAD+ —————

Fructose 6-phosphate 2 pyruvate 2ATP 2NADH 2H+

ATP

ADP
The Pentose Phosphate Pathway
Fructose 1,6-bisphosphate
A second pathway, the pentose phosphate or hexose monophos-
Aldolase phate pathway may be used at the same time as the glycolytic
Glyceraldehyde-3- P Dihydroxyacetone- P pathway or the Entner-Doudoroff sequence. It can operate either
NAD
+
Pi aerobically or anaerobically and is important in biosynthesis as
well as in catabolism.
+
NADH +H Three-carbon stage The pentose phosphate pathway begins with the oxidation of
1,3-bisphosphoglycerate glucose 6-phosphate to 6-phosphogluconate followed by the oxida-
ADP tion of 6-phosphogluconate to the pentose ribulose 5-phosphate and
Substrate-level phosphorylation CO2 (figure 9.6 and appendix II). NADPH is produced during these
ATP
oxidations. Ribulose 5-phosphate is then converted to a mixture of
3-phosphoglycerate three- through seven-carbon sugar phosphates. Two enzymes
unique to this pathway play a central role in these transformations:
(1) transketolase catalyzes the transfer of two-carbon ketol groups,
2-phosphoglycerate
and (2) transaldolase transfers a three-carbon group from sedohep-
H 2O tulose 7-phosphate to glyceraldehyde 3-phosphate (figure 9.7). The
Phosphoenolpyruvate overall result is that three glucose 6-phosphates are converted to two
ADP fructose 6-phosphates, glyceraldehyde 3-phosphate, and three CO2
Substrate-level phosphorylation molecules, as shown in the following equation.
ATP
Pyruvate
3 glucose 6-phosphate 6NADP+ 3H2O —————
2 fructose 6-phosphate glyceraldehyde 3-phosphate
3CO2 6NADPH 6H+

These intermediates are used in two ways. The fructose 6-

Figure 9.5 Glycolysis. The glycolytic pathway for the breakdown of
glucose to pyruvate. The two stages of the pathway and their products phosphate can be changed back to glucose 6-phosphate while
are indicated. glyceraldehyde 3-phosphate is converted to pyruvate by glycolytic
enzymes. The glyceraldehyde 3-phosphate also may be returned
to the pentose phosphate pathway through glucose 6-phosphate
formation. This results in the complete degradation of glucose 6-
to produce ATP. This synthesis of ATP is called substrate-level phosphate to CO2 and the production of a great deal of NADPH.
phosphorylation because ADP phosphorylation is coupled with
the exergonic breakdown of a high-energy substrate molecule. Glucose 6-phosphate 12NADP+ + 7H2O —————
ATP as an energy currency (pp. 155, 157) 6CO2 12NADPH 12H+ Pi
A somewhat similar process generates a second ATP by The pentose phosphate pathway has several catabolic and an-
substrate-level phosphorylation. The phosphate group on 3- abolic functions that are summarized as follows:
phosphoglycerate shifts to carbon two, and 2-phosphoglycerate is
dehydrated to form a second high-energy molecule, phospho- 1. NADPH from the pentose phosphate pathway serves as a
enolpyruvate. This molecule donates its phosphate to ADP forming source of electrons for the reduction of molecules during
a second ATP and pyruvate, the final product of the pathway. biosynthesis.
The glycolytic pathway degrades one glucose to two pyruvates 2. The pathway synthesizes four- and five-carbon sugars for a
by the sequence of reactions just outlined. ATP and NADH are also variety of purposes. The four-carbon sugar erythrose
produced. The yields of ATP and NADH may be calculated by con- 4-phosphate is used to synthesize aromatic amino acids and
sidering the two stages separately. In the six-carbon stage two ATPs vitamin B6 (pyridoxal). The pentose ribose 5-phosphate
are used to form fructose 1,6-bisphosphate. For each glyceralde- is a major component of nucleic acids, and ribulose
hyde 3-phosphate transformed into pyruvate, one NADH and two 1,5-bisphosphate is the primary CO2 acceptor in
ATPs are formed. Because two glyceraldehyde 3-phosphates arise photosynthesis. Note that when a microorganism is growing
 Prescott−Harley−Klein: III. Microbial Metabolism 9. Metabolism: Energy © The McGraw−Hill
Microbiology, Fifth Edition Release and Conservation Companies, 2002

Figure 9.6 The Pentose Phosphate Pathway. + + + +

3NADP 3NADPH + 3H 3NADP 3NADPH + 3H
The conversion of three glucose 6-phosphate
molecules to two fructose 6-phosphates and a 3 glucose-6- P 3 6-phosphogluconate 3 ribulose-5- P
glyceraldehyde 3-phosphate is traced. The fructose
3H2O 3CO2
6-phosphates are changed back to glucose
6-phosphate. The glyceraldehyde 3-phosphate
can be converted to pyruvate or combined with a
molecule of dihydroxyacetone phosphate (from the
glyceraldehyde 3-phosphate formed by a second
Ribose-5- P Xylulose-5- P
turn of the pathway) to yield fructose 6-phosphate.

Transketolase

Glyceraldehyde-3- P Sedoheptulose-7- P

Transaldolase

Fructose-6- P Erythrose-4- P Xylulose-5- P

Transketolase

Fructose-6- P Glyceraldehyde-3- P

Pi

Fructose-6- P Fructose-1,6-bis P Pyruvate

Figure 9.7 Transketolase and Transaldolase. Examples of The transketolase reaction

the transketolase and transaldolase reactions of the pentose
phosphate pathway. The groups transferred in these reactions O H
CH2OH
are in color.
CH2OH C C O O H
C O H C OH HO C H C

HO C H + H C OH H C OH + H C OH

H C OH H C OH H C OH CH2O P

CH2O P CH 2O P H C OH Glyceraldehyde
3-phosphate
Xylulose Ribose CH2O P
5-phosphate 5-phosphate
Sedoheptulose
7-phosphate

The transaldolase reaction

CH2OH CH2OH
C O C O
H O O H
HO C H C HO C H C

H C OH + H C OH H C OH + H C OH
H C OH CH 2O P H C OH H C OH
H C OH Glyceraldehyde CH2O P CH2O P
3-phosphate
CH2O P Fructose Erythrose
6-phosphate 4-phosphate
Sedoheptulose
178 7-phosphate
 Prescott−Harley−Klein: III. Microbial Metabolism 9. Metabolism: Energy © The McGraw−Hill
Microbiology, Fifth Edition Release and Conservation Companies, 2002

9.3 Fermentations 179

on a pentose carbon source, the pathway also can supply

Glucose
carbon for hexose production (e.g., glucose is needed for ATP
peptidoglycan synthesis).
ADP
3. Intermediates in the pentose phosphate pathway may be used Glucose 6-phosphate
+
to produce ATP. Glyceraldehyde 3-phosphate from the NADP
pathway can enter the three-carbon stage of the glycolytic NADPH
pathway and be converted to ATP and pyruvate. The latter 6-phosphogluconate
may be oxidized in the tricarboxylic acid cycle to provide
H 2O
more energy. In addition, some NADPH can be converted to
2-keto-3-deoxy-6-phosphogluconate
NADH, which yields ATP when it is oxidized by the electron
transport chain. Because five-carbon sugars are intermediates
in the pathway, the pentose phosphate pathway can be used Pyruvate Glyceraldehyde 3-phosphate
+
to catabolize pentoses as well as hexoses. NAD

NADH
Although the pentose phosphate pathway may be a source of
ADP
energy in many microorganisms, it is more often of greater im- ATP
portance in biosynthesis. Several functions of the pentose phos-
phate pathway are mentioned again in chapter 10 when biosyn-
thesis is considered more directly.

The Entner-Doudoroff Pathway ADP

Although the glycolytic pathway is the most common route for the ATP
conversion of hexoses to pyruvate, another pathway with a similar Pyruvate

role has been discovered. The Entner-Doudoroff pathway begins

with the same reactions as the pentose phosphate pathway, the for-
mation of glucose 6-phosphate and 6-phosphogluconate (figure 9.8 Figure 9.8 The Entner-Doudoroff Pathway. The sequence leading
from glyceraldehyde 3-phosphate to pyruvate is catalyzed by enzymes
and appendix II). Instead of being further oxidized, 6-
common to the glycolytic pathway.
phosphogluconate is dehydrated to form 2-keto-3-deoxy-6-
phosphogluconate or KDPG, the key intermediate in this pathway.
KDPG is then cleaved by KDPG aldolase to pyruvate and glycer-
aldehyde 3-phosphate. The glyceraldehyde 3-phosphate is con- 1,3-bisphosphoglycerate (figure 9.5) must still be oxidized back
verted to pyruvate in the bottom portion of the glycolytic pathway. to NAD . If NAD is not regenerated, the oxidation of glycer-
If the Entner-Doudoroff pathway degrades glucose to pyruvate in aldehyde 3-phosphate will cease and glycolysis will stop. Many
this way, it yields one ATP, one NADPH, and one NADH per glu- microorganisms solve this problem by slowing or stopping pyru-
cose metabolized. vate dehydrogenase activity and using pyruvate or one of its de-
Most bacteria have the glycolytic and pentose phosphate rivatives as an electron and hydrogen acceptor for the reoxidation
pathways, but some substitute the Entner-Doudoroff pathway for of NADH in a fermentation process (figure 9.9). This may lead
glycolysis. The Entner-Doudoroff pathway is generally found in to the production of more ATP. The process is so effective that
Pseudomonas, Rhizobium, Azotobacter, Agrobacterium, and a few some chemoorganoheterotrophs do not carry out respiration even
other gram-negative genera. Very few gram-positive bacteria have when oxygen or another exogenous acceptor is available. There
this pathway, with Enterococcus faecalis being a rare exception. are many kinds of fermentations, and they often are characteris-
tic of particular microbial groups (figure 9.10). A few more com-
mon fermentations are introduced here, and several others are dis-
1. Summarize the major features of the glycolytic pathway, the
cussed at later points. Two unifying themes should be kept in
pentose phosphate pathway, and the Entner-Doudoroff sequence.
Include the starting points, the products of the pathways, the
mind when microbial fermentations are examined: (1) NADH is
critical or unique enzymes, the ATP yields, and the metabolic oxidized to NAD , and (2) the electron acceptor is often either
roles each pathway has. pyruvate or a pyruvate derivative. In fermentation the substrate is
2. What is substrate-level phosphorylation? partially oxidized, ATP is formed by substrate-level phosphory-
lation only, and oxygen is not needed.
Many fungi and some bacteria, algae, and protozoa ferment
sugars to ethanol and CO2 in a process called alcoholic fermen-
9.3 Fermentations tation. Pyruvate is decarboxylated to acetaldehyde, which is then
reduced to ethanol by alcohol dehydrogenase with NADH as the
In the absence of aerobic or anaerobic respiration, NADH is not electron donor (figure 9.10, number 2). Lactic acid fermentation,
oxidized by the electron transport chain because no external elec- the reduction of pyruvate to lactate (figure 9.10, number 1), is even
tron acceptor is available. Yet NADH produced in the glycolytic more common. It is present in bacteria (lactic acid bacteria, Bacil-
pathway during the oxidation of glyceraldehyde 3-phosphate to lus), algae (Chlorella), some water molds, protozoa, and even in