7th lecture Microbial Physiology

Metabolism

Assist. Prof. Dr. Thanaa Rasheed

Metabolism:
The sum of the biochemical reactions required for energy
generation and the use of energy to synthesize cell material from
nutrient substances as small molecules in the environment.
Two components:

• Anabolism - biosynthesis
– Is the term used to describe reactions that building complex
molecules from simple ones
– requires ENERGY (ATP)

• Catabolism - degradation
– Breaking down complex molecules into simple ones
– Generates ENERGY (ATP)
Catabolic reactions produce energy as ATP adenosine
triphosphate , which can be utilized in anabolic reactions to
build cell material from nutrients in the environment.
 The differences between bacteria and
eukaryotic metabolism

 Speed: Bacteria metabolize at a rate 10 to 100 times faster.

 Versatility: Bacteria use more varied compounds as energy sources and are

much more diverse in their nutritional requirements.

 Simplicity: The prokaryotic body plan makes it possible to synthesize

macromolecules in a streamlined way.

 Uniqueness: Some biosynthetic processes, such as those producing

peptidoglycan, lipopolysaccharide, and toxins, are unique to bacteria.

 ?Why is energy needed
Like all other living things, microorganisms need to
acquire energy in order to survive. Energy is required:
1. To maintain the structural integrity of the cell by
repairing any damage to its constituents.
2. To synthesize new cellular components such as nucleic
acids, polysaccharides and enzymes.
3. To transport certain substances into the cell from its
surroundings.
4. For the cell to grow and multiply.
5. For cellular movement.
Transport of substrates
1. Fueling Reactions
Fueling reactions provide the cell with energy and with the 12
precursor metabolites used in biosynthetic reactions. No energy is
involved in this process
• The first step is the capture of nutrients from the environment. the
substrates enter despite permeability barriers by simple diffusion.
• Some transport occurs by facilitated diffusion in which a protein
carrier in the cell membrane, specific for a given compound,
participates in the shuttling of molecules of that substance from
one side of the membrane to the other. Because there is no energy
input, molecules continue to enter only as long as their
concentration is greater on the outside.
2. Active transport mechanisms
• The process involve specific protein molecules as carriers of particular
solutes, is energy linked and can establish a concentration gradient. That
is, active transport can pump "uphill.“
Bacterial systems of active transport
1. Some of which involve ATP-dependent binding proteins .
2. Others that require proton pumps driven by electron transport within
the energized cell membrane.
3. Another mechanism called group translocation involves the chemical
conversion of the solute into another molecule as it is transported.
eg: Bacterial secrete siderophores (iron-specific chelators) to trap Fe 3+ in
human blood or other body fluids because it is sequestered by iron-
binding proteins (eg, transferrin in blood and lactoferrin in secretions);
the iron-containing chelator is then transported into the bacterium by
specific active transport. Iron needed in small amounts for growth and
virulence.
 METABOLIC DIVERSITY

 Bacterial metabolism is classified into

nutritional groups on the basis of three major
criteria:
1. Source of energy, used for growth
2. Source of carbon
3. Sours of electron donors used for growth.
1. ENERGY SOURCE
a. Phototrophs —can use light energy source
b. Chemotrophs —must obtain energy from oxidation-
reduction of external chemical compounds.Use inorganic
and organic chemicals.
 2. CARBON SOURCE

a. Autotrophs —can draw carbon from carbon

dioxide

b. Heterotrophs —carbon from organic compounds

c. Mixotrophic – carbon is obtained from both organic

compounds and by fixing carbon dioxide
 These requirements can be
combined:
1. Photoautotrophs - light energy, carbon
from carbon dioxide
2. Photoheterotrophs —light energy, carbon
from organic compounds
3. Chemoautotrophs —energy from chemical
compounds, carbon from CO2
4. Chemoheterotrophs —energy from
chemical compounds, carbon from organic
compounds
 CHEMOHETEROTROPHS
 Specifically, their energy source is electrons from
hydrogen atoms in organic compounds.

      Saprophytes—live on dead organic matter

      Parasites—nutrients from a living host

 This group (more precisely chemoorganoheterotrophic)

includes most bacteria as well as all protozoa,
fungi, and animals. All microbes of medical
importance are included in this group.
Enzymes: is a cellular catalyst , specific to a particular reaction or group
of reactions. It is central to the metabolic processes of any cell. Nothing
works without enzymes!

• How important are enzymes?

– all chemical reactions in living organisms require
enzyme
enzymes to work
• building molecules +
– synthesis enzymes
enzyme
• breaking down molecules +
– digestive enzymes
– enzymes speed up reactions
• “catalysts”
 Enzyme properties
• Enzymes are proteins or RNA that catalyze most biochemical reactions
by forming enzyme-substrate complexes.
• The binding of the substrate by an enzyme makes possible both bond-
forming and bond-breaking reactions, depending on the pathway
involved. Enzymes may utilize cofactors as carriers and activators.
• Enzymes are classified and named according to the kinds of reactions
they catalyze.
• To function effectively, enzymes require specific conditions of
temperature, pH, and osmotic pressure.
• Enzyme activity is regulated by processes that affect the enzymes
directly, or by altering the amount of enzyme produced during protein
synthesis.
Reduction – Oxidation reactions (Redox)

What is “redox” to a microbe?

Redox: is short for reduction – oxidation reaction
used by microbes to generate energy involve the
transfer of electrons from one molecule to another.
• When a molecule (or atom or ion) loses an electron, it
is said to be oxidized. (Note, that despite the
terminology, oxygen does not necessarily take part in
the reaction).
• Conversely, when an electron is gained, the recipient
is reduced .
Many metabolic reactions involve the loss of a
hydrogen atom; since this contains one proton and
one electron, the reaction is regarded as an
oxidation, because an electron has been lost:

The lactate in the example above, by losing two

hydrogen atoms, has automatically lost two electrons
and thus become oxidised to pyruvate.
Two important molecules, the coenzymes nicotinamide
adenine dinucleotide (NAD+) and nicotinamide adenine
dinucleotide phosphate (NADP+).
Both are derivatives of the B vitamin niacin, and each can
exist in an oxidized and a reduced form:
Coupled reaction
In the oxidation of lactate, the oxidizing power is
provided by the reduction of NAD+, so the full
story would be:

As the lactate is oxidized, so the NAD+ in the

coupled reaction is reduced. It is said to act as the
electron acceptor; there can be no oxidation
without reduction, and vice versa.
 Energy Generating Patterns
 Respiration is the term used to describe those ATP
generating processes, aerobic or anaerobic, by which
oxidation of a substrate occurs, with an inorganic
substance acting as the final electron acceptor. In
aerobic respiration, that substance is oxygen; in anaerobic
respiration, a substance such as nitrate or sulphate can
fulfil the role.
 After Sugars are made or obtained, they are the energy
source of life.
 Breakdown of sugar(catabolism) different ways:
Aerobicrespiration
Anaerobic respiration

Fermentation
 Aerobic respiration
 Most efficient way to extract energy from glucose, that
converts glucose to CO 2 , produces H 2 O, and generate
energy. It relies on free oxygen as the final acceptor for
electrons and hydrogens and generates a relatively large
amount of ATP.
C6H12O6 + 6O2 −→ 6CO2 + 6H2O
Process:
1. Glycolysis also called the Embden-Meyerhof- Parnas
(EMP) pathway
2. Kreb Cycle also known as the citric acid or tricarboxylic
acid cycle
3. Electron transport chain( the respiratory chain ,electron
transport and oxidative phosphorylation).
Using oxygen (1/2 O2) in metabolism
creates toxic waste.
Microbes that are able to use aerobic
respiration produce enzymes to detoxify
oxygen:

Catalase: H2O2 ------- H20 and 02

Superoxide dismutase (SOD): oxygen radical -- H20 and O2

Microbes that don’t make these enzymes cannot

exist in the presence of oxygen.
Glycolysis (EMP pathway)
It can be defined as the initial sequence of reactions, in which a
molecule of glucose is converted to two molecules of pyruvate.

In the first phase of glycolysis, glucose is phosphorylated and its

six carbon ring structure rearranged, before being cleaved into
two three-carbon molecules.

In the second phase, each of these undergoes oxidation, resulting

in pyruvate.

Glycolysis takes place in the cytoplasm, comprises a series of 10

linked reactions, with a net gain of two molecules of ATP.
In terms of energy, glycolysis can be divided into a ‘sowing’
phase, in which two molecules of ATP are expended per molecule
of glucose, followed by a ‘reaping’ phase which yields four
molecules of ATP. The overall energy balance is therefore a gain
of two molecules of ATP for each molecule of glucose oxidized to
pyruvate.

In addition, the second phase features the conversion of two

molecules of NAD+ to NADH.

The reactions by which ATP is generated from ADP in the second

phase of glycolysis are examples of substrate-level
phosphorylation, so-called because the phosphate group is
transferred directly from a donor molecule.
 PRINCIPLES OF ENERGY GENERATION

 Glycolysis. Two molecules of ATP are ‘spent’ in the first stage of glycolysis, in which glucose
is converted into the 3-carbon compounds glyceraldehyde 3-phosphate and dihydroxyacetone
phosphate. During the second stage, four ATPs are produced per molecule of glucose, so there
is a net gain of two ATPs. In addition, reducing power is generated in the form of NADH (two
molecules per molecule of glucose).
The overall energy balance is therefore a gain of
two molecules of ATP for each molecule of glucose
oxidized to pyruvate. In addition, the second phase
features the conversion of two molecules of NAD+
to NADH.

GLYCOLYSIS 2 ATP + 2NADH

Carbohydrate Catabolism

• Alternatives to Glycolysis
Two pathways
– Pentose phosphate pathway
– Entner-Doudoroff pathway
– Yield fewer molecules of ATP than glycolysis
– Reduce coenzymes and yield different metabolites needed in
anabolic pathways

.Copyright © 2011 Pearson Education Inc

Aerobic respiration
We shall now examine the fate of the pyruvate
produced as the end-product of glycolysis, this
depends on whether the organism in question is
aerobic or anaerobic.

During glycolysis, NAD+ was reduced to NADH.

In order for glucose metabolism to continue, this
supply of NAD+ must be replenished; this is
achieved either by respiration or fermentation.
In most aerobic organisms, the pyruvate is
completely oxidized to CO2 and water by entering
the tricarboxylic acid (TCA) cycle, also known as
the Krebs cycle or simply the citric acid cycle.

The TCA cycle is a series of reactions that oxidize

acetate to CO2, generating reducing power in the
form of NADH and FADH2 for use in the electron
transport chain.
Pyruvate does not itself directly participate in the
TCA cycle, but must first be converted into the
two-carbon compound acetyl-Coenzyme A:

This step results in the formation of 2NADH

• For each ‘turn’ of the citric acid cycle, one
molecule of ATP, three molecules of NADH
and one molecule of FADH2 are produced
(FADH2 is the reduced form of another
coenzyme, FAD).

• TCA Cycle 2ATP + 6NADH +

2FADH2
Krebs Cycle (Citric Acid
Cycle,TCA)
 Series of chemical reactions that begin and end with citric
acid

1. Initial substrate – modified end product of Glycolysis

 2 Pyruvic Acid is modified to 2Acetyl-CoA, which enters
the TCA cycle
2. Circuit of organic acids – series of oxidations and reductions
 Prokaryotes – Cytoplasm of bacteria & Cell Membrane

 Products:
 2 ATP
 6 NADH2
 2 FADH2
 4 CO2
• Most of the energy originally stored in the glucose
molecule is now held in the form of the reduced
coenzymes (NADH and FADH2) produced during
glycolysis and the TCA cycle.
• This is now converted to no less than 34 molecules
of ATP per glucose molecule by oxidative
phosphorylation in the remaining steps in aerobic
respiration.
• In the final phase of aerobic respiration, electrons
are transferred from NADH and FADH2, via a
series of carrier molecules known collectively as
the electron transport (or respiratory) chain to
oxygen, the terminal electron acceptor.
Electron transport

 The electron transport chain is a series of

donor/ acceptor molecules that transfer
electrons from donors (e.g. NADH) to a
terminal electron acceptor (e.g. O2).
 Sufficient energy is released at three
points in the electron transport chain for
the transfer of protons to the outside of
the membrane, resulting in a gradient of
both concentration and charge (proton
motive force).
• The protons are able to return across the membrane and
achieve an equilibrium through specific protein channels
within the enzyme ATPsynthase. The energy released by the
protons as they return through these channels enables the
ATP synthase to convert ADP to ATP

• Aerobic respiration in eucaryotes is slightly less efficient than

in procaryotes due to the fact that the three stages take place
at separate locations. Thus the total number of ATPs
generated is 36 rather than the 38 in procaryotes
The electron transport chain. Electrons from NADH and FADH2 pass from one electron
carrier to another, with a gradual release of energy as ATP by chemiosmosis. The electron
carriers are arranged in order of their reduction potential (tendency to gain electrons) and
oscillate between the oxidised and the reduced state.
• How does this transfer of electrons lead to the formation of
ATP?
• The chemiosmotic theory proposed by Peter Mitchell in
1961 offers an explanation by pumping protons across a
membrane. 
Yield of ATP by aerobic respiration in procaryotes
Fermentation

 We turn now to the fate of pyruvate when oxygen is not

available for aerobic respiration to take place.
 Microorganisms are able, by means of fermentation, to
oxidise the pyruvate incompletely to a variety of end
products.
Fermentation: a microbial process by which an organic
substrate (usually a carbohydrate) is broken down without
the involvement of oxygen or an electron transport chain,
generating energy by substrate level phosphorylation.
• Two common fermentation pathways result in the
production of respectively lactic acid and ethanol.
 Fermentation
 Incomplete oxidation of glucose or other
carbohydrates in the absence of oxygen

 Uses organic compounds as terminal electron

acceptors

 Effect - a small amount of ATP

 Production of ethyl alcohol by yeasts acting on

glucose

 Formation of acid, gas & other products by the

action of various bacteria on pyruvic acid
 Fermentation
 Fermentation pathways
a. Homolactic acid F.
P.A -----> Lactic Acid
eg. Streptococci, Lactobacilli
b.Alcoholic F.
P.A -----> Ethyl alcohol
eg. yeast
 Anaerobic respiration
Utilizes same 3 coupled pathways as Aerobic Respiration
Used as an alternative to aerobic respiration
Final electron acceptor something other than oxygen:
NO3- : Pseudomonas, Bacillus.
SO4-: Desulfovibrio
CO3-: methanogens

In Facultative organisms
In Obligate anaerobes

Lower production of ATP because only part of the TCA

cycle and the electron transport chain operate.
• Anaerobic respiration is not as productive in terms
of ATP production, because electron acceptors such
as nitrate or sulphate have less positive redox
potentials than oxygen.
• Anaerobic respiration is not the same as
fermentation. The latter process does not involve the
components of the electron transport chain and
much smaller amounts of energy are generated.
THANK YOU