Microbial Metabolism

Why learn about microbial metabolism?

It is important to have a basic API20E test strip
understanding of metabolism
because it governs the survival
and growth of microorganisms.

Good metabolic function makes

pathogens more successful at
causing disease.

Metabolic functions can be used

in diagnosis MacConkeys media with Salmonella growing on left plate and
E. coli on right
 Major Categories of Metabolic Reactions
Catabolic processes harvest the energy released during the
breakdown of compounds and use it to make ATP. They also
make precursor metabolites used in biosynthesis.

Anabolic processes (biosynthesis) synthesize and assemble

subunits of macromolecules that make up the cell structures.
They use the ATP and precursor metabolites produced in
catabolism.
Both processes involve electron transfer and oxidation and
reduction reactions
Major Categories of
Metabolic Reactions
 Electron Transfer

Energy is released when electrons are

moved from an energy sources with a
low affinity for electrons (less
electronegative; electron donor) to
a terminal electron acceptor with a
higher affinity (more electronegative;
electron acceptor)
Energy Generated in Electron Transfer
Stored as ATP
H H
N
Adenine
N
N
O- O- O- H N
N
O- P O P O P O C H
O
O O O

Ribose
H H H H

OH OH
Adenosine monophosphate (AMP)
Adenosine diphosphate (ADP)
Adenosine triphosphate (ATP)
Bond that releases energy
when broken
 Phosphorylation stores energy

P
Adenosine P P + Energy
Adenosine P P P
ADP
ATP
 Dephosphorylation releases
energy
P + Energy
Adenosine P P P Adenosine P P

ATP
ADP
 ATP produced by:

Substrate-level phosphorylation

Oxidative Phosphorylation
 Oxidative Phosphorylation
Phosphate (pi) is added Outside cell
to ADP by turning the
wheel of ATP synthase

Inside cell
H+are used to turn ATP
Synthase, powering ATP
production
 Substrate Level Phosphorylation
Instead of using ATP Synthase, ATP is produced using an
enzyme reaction to transfer a phosphate to ADP
 Substrate Level Phosphorylation
For example, in glycolysis, an enzyme moves the phosphate
from phosphoenolpyruvate onto the ADP .
 Oxidation and Reduction Reactions

An oxidation reaction is a chemical reaction in which a

molecule loses one or more electrons.

A reduction reaction is a chemical reaction in which a

molecule gains one or more electrons.

Oxidation and reduction reactions always occur together:

Redox reactions
 Redox Reactions

When a substance is reduced, it gains electrons.

OIL RIG

When a substance is oxidized, it loses electrons

Biological Redox Reaction
Reduction

15

Oxidation
 Question
The molecule that is oxidized in a redox reaction

A. gains electrons and gains potential energy.

B. loses electrons and loses potential energy.
C. gains electrons and loses potential energy.
D. loses electrons and gains potential energy.
E. neither gains nor loses electrons, but gains or
loses potential energy.
 Electron Carrier Molecules
Substrate I Final electron acceptor
NAD+

NADH
Substrate I + ATP

Reducing Power: electrons available in reduced

electron carriers
 Question
Is NAD+ an oxidized or reduced electron
carrier?
A.Oxidized
B.Reduced
 Question
When electrons move closer to a more electronegative
atom, what happens?
A. The more electronegative atom is reduced, and energy is
released.
B. The more electronegative atom is reduced, and energy is
consumed.
C. The more electronegative atom is oxidized, and energy is
consumed.
D. The more electronegative atom is oxidized, and energy is
released.
 Overview of Glucose Respiration
G Glucose
L
Y
C
O
Fermentation
Catabolism L
Y
SI
S

(1)Oxidize glucose to generate: Pyruvate

2 Pyruvate
ATP, reducing power and (or derivative)

precursor metabolites
Formation of
Acetyl-CoA fermentation
end-products
(2) Transfer the electrons KREBS
carried by NADH and FADH2 CYCLE

(reducing power) to the

Electrons
terminal electron acceptor
 Glycolysis
Glucose

Occurs in cytoplasm of Glucose 6-phosphate 1,3-Bisphosphoglyceric acid

eukaryotes and
prokaryotes Fructose 1,6-bisphosphate 3-Phosphoglyceric acid

Oxygen is not involved

Dihydroxyacetone
phosphate (DHAP) Phosphoenolpyruvic acid (PEP

Glyceraldehyde 3-phosphate (G3P) Pyruvic acid

 Glycolysis
Oxidize 1 glucose molecule to generate:
1. ATP: 2 ATP (net) by substrate-level
phosphorylation
2. Reducing power: 2 NADH
3. Precursor Metabolites: 2 pyruvate molecules +
5 other precursor metabolites (intermediates)
 Fates of Pyruvate
Glycolysis

Amino acids Pyruvic

Sugars acid
Fat metabolites Acetaldehyde

Acetyl CoA Alcohol

Acids, gas Acetone
Krebs 2, 3 butanediol
cycle
Anabolic
pathways Respiration Fermentation
 Cellular Respiration Respiration G Glucose Fermentation
L Y
C
O
L
In metabolism, respiration occurs Y
SI
S

at the cellular level and is not the 2 Pyruvate Pyruvate

same as breathing (respiration at (or derivative)

the macroscopic level).

Formation of
Acetyl-CoA fermentation
end-products
Pyruvic acid becomes completely KREBS
oxidized CYCLE

Electrons
 3 Stages of Cellular Respiration

1. Transition step: Synthesis of acetyl-CoA

2. Krebs cycle
3. Electron Transport Chain - final series of
redox reactions
 Synthesis of Acetyl CoA Pyruvic acid

Yield (from 1 glucose/ 2 pyruvates): Decarboxylation

1. Reducing power: 2 NADH

2. Precursor metabolites: 2 acetyl Acetate

CoA Coenzyme A

Acetyl-coenzyme A
(acetyl-CoA)
Respiration Fermentation

Acetyl-CoA

Citric acid
Oxaloacetic
acid
Malic acid
Isocitric acid
TCA
Fumaric acid CYCLE
-Ketoglutaric acid
Succinic acid
Succinyl-CoA
 TCA Cycle (aka Krebs cycle)
Yield (from 1 glucose/ 2 acetyl CoA):
1. ATP: 2 ATP by substrate-level phosphorylation
2. Reducing power: 6 NADH and 2 FADH2

Location:
Occurs in cytosol of prokaryotes and in matrix of
mitochondria in eukaryotes
 Electron Transport Chain and ATP Synthesis
2 main steps:
1) Energy from electrons
used to pump protons (H+)
across the membrane,
establishing a proton
gradient

2) The enzyme, ATP

synthase, uses the energy
of the proton gradient to
drive the synthesis of ATP
by oxidative
phosphorylation
 Generating a Proton Gradient
Some electron carriers
function also as proton
pumps
 Location of the ETC
Eukaryotes: inner mitochondrial membrane of
eukaryotes
Prokaryotes: cytoplasmic membrane of prokaryotes
 Synthesis of ATP via Chemiosmosis
Chemiosmosis: Use of electrochemical gradients to
generate ATP

Protons flow down electrochemical gradient through

ATP synthases that phosphorylate ADP to ATP

ATP generated via oxidative phosphorylation

 Synthesis of ATP via Chemiosmosis
FMN Ubiquinone
Phospholipid Cyt b Cyt a3
membrane Cyt c Cyt a

NADH from Cyt c1

glycolysis, ATP synthase
Krebs cycle, FADH2 from
Krebs cycle
O2
Cytoplasm of prokaryote
or matrix of mitochondrion
 Question
In which direction would electrons flow?
In which direction would proton pumping go?
 ATP Yield of Aerobic
Respiration in Prokaryotes

38 ATP
(yield from ETC= 34)

Most significant ATP

yield is from ETC
 Anaerobic Respiration
Use of another compound than O2 as final electron acceptor
in the ETC
Examples
Nitrate ion NO3- to NO2-
Pseudomonas, E coli, Bacillus
Sulfate ion SO42- to H2S
Methanogens
Carbonate ion CO32- to CH4
Methanogens
 Aerobic vs Anaerobic (a) AEROBIC RESPIRATION (b) ANAEROBIC RESPIRATION

Respiration Glycolysis
Glucose (6 C)
Glycolysis
Glucose (6 C)

Aerobic respiration: ATP

NADH
2 pyruvate (3 C)
ATP
NADH
2 pyruvate (3 C)

Obligate aerobes, microaerophiles CO2 CO2

and facultative anaerobes in Acetyl Co A Acetyl Co A

presence of O2 FADH2 FADH2

CO2 CO2

Oxygen
Krebs Krebs
NADH NADH

ATP ATP

Anaerobic respiration: Electrons

Electron transport
Electrons
Electron transport

facultative anaerobes in absence of

O2 O2 is final electron
acceptor.
Nonoxygen electron acceptors
(ie SO42-, NO3-,CO32-)

Inorganic oxygen-containing Maximum Maximum ATP yield is

variable
ATP produced =
molecule 38
depending on final electron
acceptor (<38)
 Fermentation
Obligate anaerobes, aerotolerant, facultative anaerobes
Does not require oxygen
Used by microbial organisms that cannot respire
o a suitable inorganic terminal electron acceptor is not
available
o they lack an electron transport chain

ATP yield= 2 ATP by substrate phosphorylation (glycolysis)

 Fermentation
FERMENTATION
Glycolysis
Glucose (6 C)
ATP
NADH
2 pyruvate (3 C)
CO2 Regenerates oxidized
Fermentation
electron carriers (NAD+)
Lactic acid Acetaldehyde
for subsequent rounds of
Ethanol
1CO2 glycolysis
Or other alcohols,
acids, gases

Pyruvate or a derivative is final

electron acceptor
Maximum
ATP produced = 2
Fermentation End products
Major Catabolic
Pathways
 Question (a) AEROBIC RESPIRATION
Glycolysis
(b) ANAEROBIC RESPIRATION
Glycolysis
FERMENTATION

Glycolysis

Glucose
(6 C)
Glucose (6 C) Glucose (6 C)

Which metabolic ATP ATP

ATP
NADH
NADH NADH 2 pyruvate
(3 C)
pathways can humans 2 pyruvate (3 C) 2 pyruvate (3 C)

CO2
perform (check all that CO2 CO2
Fermentation
Acetyl Co A Acetyl Co A
apply)?
FADH2 FADH2
Lactic acid
Acetaldehyde
Krebs CO2 Krebs CO2
NADH NADH

a. Aerobic respiration ATP ATP

Ethanol
1CO2

b. Anaerobic respiration Electrons Electrons

c. Fermentation
d. None of the above
Or other alcohols,
acids, gases
Electron transport Electron transport
O2 is final electron Nonoxygen electron acceptors
Pyruvate or a derivative is
electron acceptor
acceptor. (ie SO42-, NO3-,CO32-)
Maximum Maximum
ATP produced =
Maximum
ATP produced = <38 Variable depending on ATP produced = 2
38 final electron acceptor
 Anabolic Pathways
Enzymes Cell wall Membranes
Chromosomes Cell
Membranes Storage Storage
structure
Anabolism

Nuclei Starch Lipids

c acids Proteins Cellulose Fats Macromolecule

Nucleotides Amino acids Carbohydrates Fatty acids Building

block
 Anabolic Pathways

require energy
in the form of
ATP and
precursor
metabolites