Course: Medical Microbiology, MBIM 650/720

Lecture 26: Nutrition, Growth and Energy Metabolism.

Faculty: Dr. Fox, Phone: 733 3288, E-mail: afox@med.sc.edu, Office: C-19, Blg. 2

Suggested reading: Murray, Fourth Edition, Chapter 4 2002/2003

KEY WORDS

Obligate aerobe Growth curve
Obligate anaerobe Glycolysis
Aerotolerant anaerobe Fermentation
Facultative anaerobe Anaerobic respiration
Microaerophilic Aerobic respiration
Siderophore Tricarboxylic acid (TCA) cycle
Mesophile or Krebs cycle
Thermophile Oxidative phosphorylation
Psychrophile Ubiquinone
Generation time Glyoxylate pathway

Bacterial requirements for growth include oxygen, (or its absence) nutrients, optimal
temperature and pH.

Oxygen Requirements: Obligate aerobes must grow in the presence of oxygen; they can
not carry out fermentation. Obligate anaerobes do not carry out oxidative phosphorylation.
Furthermore, they are killed by oxygen; they lack certain enzymes (e.g. catalase [breaks
hydrogen peroxide, H
2
O
2,
to H
2
O and

O
2
], peroxidase (NADH + H
2
O
2
converted to NAD and

O
2
), superoxide dismutase (superoxide [O
2
-
] to H
2
O
2
) which detoxify both hydrogen peroxide
and oxygen free radicals (superoxide) produced as side-products 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 include sources of organic carbon, N, P, S and metal ions (e.g.
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). Thus bacteria that
ineffectively compete with the host for iron are poor pathogens.


1
Temperature: 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).

pH. Many bacteria grow best at neutral pH; however certain bacteria can survive and
even grow in quite acid or alkaline conditions.

Measuring bacterial mass in liquid cultures of bacteria. Common methods include a)
Turbidity (how cloudy is a liquid culture of bacterial - a measure of total bacteria (live and dead).
This is usually quantitated with a spectrophotometer or b) The number of viable bacteria in a
culture - usually assessed by counting the number of colonies that grow after streaking a known
volume on a plate (plate counting of colony forming units). In either case plotting the log of
turbidity or number of living cells versus time is referred to as the growth curve. The generation
times is defined as the time required for bacterial mass to double.
log of turbidity

Plotting the or number of living cells
versus time is referred to as the growth curve.
The generation time is the time required for bacterial mass to double.


METABOLISM OF SUGARS (as an example of metabolic pathways)

Glycolysis (Embden, Meyerhof Parnas Pathway) is the most common pathway in bacteria for
sugar catabolism (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
are stored in the newly formed compounds (ATP and NADH).


NAD NADH
Glucose ____________________________________> Pyruvate
(C6) ADP ATP (C3)
*

2


*
refers to number of carbons in molecule

There are alternatives, for catabolizing sugars, to produce stored energy within ATP
including the pentose phosphate pathway (hexose monophosphate shunt) which is found in most
animal and plant cells. NADPH is generated using this pathway. A third pathway (the Entner
Doudoroff pathway) is less often found in certain species of bacteria and is not seen in animals.


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.

NADH NAD
Pyruvate ___________________________> Short chain alcohols or
(C3) fatty 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 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)

NAD NADH
Pyruvate ___________________________________> 3 CO
2

(C3) (C1)
3


Oxidative phosphorylation

NADH NAD
O
2
______________________________________________> H
2
O
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.

a) Loss of CO
2
(C1) followed by addition to a C4 component of the cycle to produce a C6
component. Thus the number of molecules of C6 produced equals the number of molecules of
C4 initially present.

_CO
2

C3 C2

C2
C4 C6

b) By addition of CO
2
to pyruvate producing a C4 compound. In this instance additional
molecule of C4 (a cycle component) are formed.

+CO
2

C3 C4

Thus if cycle components are removed for use in other biosynthetic pathways they can be
replenished because of b). This allows sugars to be used as a sole carbon source.


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 CO
2
and
C4 regenerated. Overall no increase in the number of molecules of cycle intermediates occurs.
Thus if fatty acids are the sole carbon source then no Krebs cycle intermediates can be removed
without shutting it down:

+C2
C4 C6

Instead bacteria utilize the glyoxylate cycle (a modified Krebs Cycle) in which the
enzymatic steps in which 2 CO
2 m
olecules are removed from the C6 intermediate are by-passed.
Instead the C6 intermediate is converted to two C4 compounds (both components of the cycle).
4
Thus for every acetyl group (from fatty acids) an additional cycle intermediate can be produced.
The glyoxylate pathway is not generally found in animal cells since pre-formed fatty acids,
present in food, are utilized.

C6 C4

+
+C2
C2 C4


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.





The Krebs and Glyoxylate Cycles



-
+
C3
C2
C4
C6
C2
C2
C4
C4
C4
C5
C6

5