Bacteriology

NUTRITION, GROWTH AND

ENERGY METABOLISM
 Rationale
⚫ Bacterial requirements for growth include:
⚫ sources of energy;
⚫ "organic" carbon (e.g. sugars and fatty
acids);
⚫ metal ions (e.g. iron);
⚫ optimal temperature;
⚫ optimal pH;
⚫ the need (or lack of need for oxygen).
 Oxygen Requirements
⚫ Obligate aerobes must grow in the presence of oxygen; they
cannot carry out fermentation.
⚫ Obligate anaerobes do not carry out oxidative
phosphorylation.
⚫ Instead they are killed by oxygen, because they lack certain
enzymes such as:
⚫ catalase, which breaks down hydrogen peroxide, H2O2, to
water and oxygen;
⚫ Peroxidase, by which NADH + H2O2 are converted to NAD
and O2;
⚫ superoxide dismutase, by which superoxide, O2. is converted
to H2O2;
⚫ These enzymes detoxify peroxide and oxygen free radicals
produced during metabolism in the presence of oxygen.
⚫ Aerotolerant anaerobes are bacteria that respire
anaerobically, but can survive in the presence of
oxygen.
⚫ Facultative anaerobes can perform both
fermentation and aerobic respiration.
⚫ In the presence of oxygen, anaerobic respiration
is generally shut down and these organisms
respire aerobically.
⚫ Microaerophilic bacteria grow well in low
concentrations of oxygen, but are killed by higher
concentrations.
 Nutrient Requirements
⚫ These include sources of organic carbon,
nitrogen, phosphorus, sulfur and metal ions
including iron.
⚫ Bacteria secrete small molecules that bind iron
(siderophores, e.g. enterobactin, mycobactin).
Siderophores (with bound iron) are then
internalized via receptors by the bacterial cell.
⚫ The human host also has iron transport proteins
(e.g. transferrin). Bacteria that ineffectively
compete with the host for iron are poor
pathogens.
 Temperature and pH
⚫ Bacteria may grow at a variety of temperatures from
close to freezing to near to the boiling point of water.
⚫ Those that grow best at the middle of this range are
referred to as mesophiles; which includes all human
pathogens and opportunists.
⚫ Those having lower and higher temperature optima
are respectively known as psychrophiles and
thermophiles.
⚫ Many bacteria grow best at neutral pH;
⚫ however certain bacteria can survive and even grow
in quite acid or alkaline conditions.
Methods for measuring bacterial mass
in liquid cultures of bacteria
⚫ turbidity : the cloudiness of a liquid culture of bacteria -
a measure of total bacteria (live and dead) usually
assessed with a spectrophotometer);
⚫ "plate counting" of colony forming units: the number of
viable bacteria in a culture can be assessed by counting
the number of colonies that grow after streaking a
known volume on a plate;
⚫ in either case plotting the log of turbidity or number of
living cells versus time is referred to as the growth curve.
⚫ the generation time is defined as the time required for
bacterial mass to double.
 Glycolysis: Embden, Meyerhof and
Parnas (EMP) Pathway
⚫ This is the most common pathway in bacteria for sugar
catabolism (It is also found in most animal and plant cells).
⚫ A series of enzymatic processes result in conversion of sugars
into pyruvate, generating ATP (adenosine triphosphate) and
NADH (nicotinamide adenine dinucleotide).
⚫ Chemical energy needed for biosynthetic purposes is stored
in the newly formed compounds (ATP and NADH)

NAD ---> NADH

Glucose Pyruvate
------------------------------------>
(C6) (C3)
ADP ---> ATP
refers to number of carbons in molecule
⚫ There are alternatives to this pathway for
catabolizing sugars in order to produce
stored energy within ATP.
⚫ These include the pentose phosphate
pathway (hexose monophosphate shunt)
which is found in most animal and plant
cells. NADPH is generated using this
pathway.
⚫ Another pathway, the Entner Doudoroff
pathway, is generally only found in certain
bacterial cells.
 Anaerobic Respiration
⚫ Anaerobic respiration includes glycolysis and fermentation.
During the latter stages of this process NADH (generated
during glycolysis) is converted back to NAD by losing a
hydrogen. The hydrogen is added to pyruvate and, depending
on the bacterial species, a variety of metabolic end-products
are produced.

Short chain alcohols or (C3) fatty

NADH --> NAD
Pyruvate acids (such as lactic acid or ethanol)
------------------------->
(C2-C4)
 Aerobic Respiration
⚫ Aerobic Respiration involves glycolysis and the tricarboxylic
acid cycle (Krebs cycle).
⚫ Pyruvate is fully broken down to carbon dioxide (C1) and in
the process, NAD is converted to NADH.
⚫ Thus, in aerobic fermentation, NADH is generated from two
sources (glycolysis and the Krebs cycle).
⚫ Oxidative phosphorylation converts excess NADH back to
NAD and in the process produces more ATP (stored energy).
⚫ Ubiquinones and cytochromes are components of the
electron transport chain involved in this latter process.
⚫ Conversion of oxygen to water is the final step in the process.
 Krebs Cycle (C4-C6 intermediate
compounds)
Pyruvate NADH --> NAD
3CO2 (C1)
(C3) ------------------------->

Oxidative phosphorylation
NADH --> NAD
-------------------------> H2O
O2
ADP --> ATP
The Krebs cycle contains 4 and 6 carbon
intermediates. Pyruvate (C3) can feed
into the Krebs cycle in such a way that the
number of C4/C6 intermediates remains
the same or increases.
 Krebs Cycle
⚫ Loss of CO2 (C1) from pyruvate to form acetyl coA,
followed by addition to a C4 component of the cycle
(oxaloacetate) produces a C6 component (citric acid).
Thus the number of molecules of C6 produced equals
the number of molecules of C4 initially present.
⚫ By addition of CO2 to pyruvate a C4 compound is
produced. In this instance, additional molecules of C4 (a
cycle component) are formed.
⚫ Thus if some of the cycle components are removed for
use in other biosynthetic pathways, they can be
replenished via this reaction.
 METABOLISM OF FATTY ACIDS
⚫ Fatty acids are broken down to acetyl groups (C2)
which feed into the Krebs Cycle by addition to a
C4 intermediate to produce C6.
⚫ During the cycle, the added C2 is lost as CO2 and
C4 regenerated.
⚫ Overall, no increase in the number of molecules
of cycle intermediates occurs.
⚫ Thus, if fatty acids are the sole carbon source, no
Krebs cycle intermediates can be removed
without shutting it down.
The Krebs cycle contains 4 and 6 carbon
intermediates. Pyruvate (C3) can feed
into the Krebs cycle in such a way that the
number of C4/C6 intermediates remains
the same or increases.
⚫ Instead, bacteria utilize the glyoxylate cycle a
modified Krebs Cycle) where the enzymatic steps
in which two CO2 molecules are removed from
the C6 intermediate are by-passed.
⚫ The C6 intermediate is converted to two C4
compounds (both components of the cycle).
⚫ Thus for every acetyl group (from fatty acids), an
additional cycle intermediate can be produced.
⚫ The glyoxalate pathway is not generally found in
animal cells since pre-formed fatty acids, present
in the food, are utilized.
⚫ In summary, the Krebs Cycle functions in a
biosynthetic and energy producing fashion.
⚫ However, if intermediates are to be removed
for use in other metabolic pathways, they
must be replenished.
⚫ The replenishment process for utilization of
sugars and fatty acids is different.