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Bacterial Growth and Metabolism: Prof. Dr. Batool Hassan Al-Ghurabi

1) Bacteria grow through binary fission, which leads to exponential increases in cell numbers over time. The generation time for most bacteria is 1-3 hours. 2) Bacteria go through four growth phases: lag phase, exponential/log phase, stationary phase, and decline phase. During exponential phase, the number of bacteria doubles within each generation time. 3) Bacteria require nutrients, an appropriate temperature range, oxygen levels, pH, and osmotic pressure to grow. Their metabolic pathways and byproducts can be used to classify different bacteria.

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0% found this document useful (0 votes)
106 views63 pages

Bacterial Growth and Metabolism: Prof. Dr. Batool Hassan Al-Ghurabi

1) Bacteria grow through binary fission, which leads to exponential increases in cell numbers over time. The generation time for most bacteria is 1-3 hours. 2) Bacteria go through four growth phases: lag phase, exponential/log phase, stationary phase, and decline phase. During exponential phase, the number of bacteria doubles within each generation time. 3) Bacteria require nutrients, an appropriate temperature range, oxygen levels, pH, and osmotic pressure to grow. Their metabolic pathways and byproducts can be used to classify different bacteria.

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Bacterial Growth and Metabolism

Prof. Dr. Batool Hassan Al-Ghurabi


Growth of Microbes
 Microbial growth: Microbes grow via binary
fission, resulting in exponential increases in
numbers

 Bacterial “growth” means an increase in the


number of cells, not an increase in cell size.

 One cell becomes colony of millions of cells


 Bacteria grow by binary fission to produce
identical offspring, which cannot be distinguished
as a parent or offspring
Binary Fission
Generation Time
 Generation time; is the time it takes for a
single cell to grow and divide
 Average for bacteria is 1-3 hours
 Escherichia coli: 20 minutes……20 generations
(7 hours), 1 cell becomes 1 million cells!
 Mycobacterium much slower: (12-24h)
Fig. 7.14a
Phases of Growth
Four main growth phases
-Lag phase
-Exponential (Log) phase
-Stationary phase (Post-exponential)
-Decline phase
In lag phase, bacteria adapt themselves to growth
conditions. It is the period where the
individual bacteria are maturing and not yet able to
divide.
During the lag phase synthesis of RNA, enzymes and
other molecules occurs.
In this phase cells change very little because the cells do
not immediately reproduce in a new medium. During
this phase cells are not dormant.
The log phase (sometimes called the logarithmic
phase or the exponential phase) is a period
characterized by cell doubling.
The number of new bacteria appearing per unit time
is proportional to the present population. If growth
is not limited, doubling will continue at a constant
rate so both the number of cells and the rate of
population increase doubles with each consecutive
time period.
Exponential growth cannot continue indefinitely,
because the medium is soon depleted of nutrients
and enriched with wastes.
The stationary phase is often due to a growth-
limiting factor such as the depletion of an essential
nutrient, and/or the formation of an inhibitory
product such as an organic acid.
Stationary phase results from a situation in which
growth rate and death rate are equal.
Mutations can occur during stationary phase. DNA
damage is responsible for many of the mutations
arising in the genomes of stationary phase or
starving bacteria.
Endogenously generated reactive oxygen
species appear to be a major source of such damages.
 At death phase (decline phase), bacteria die. This
could be caused by lack of nutrients,
environmental temperature above or below the
tolerance band for the bacteria , or other injurious
conditions.
• Media for bacterial growth
•For identification of bacteria, a culture is obtained by
growing the organisms on artificial culture media.
Types of culture media
Simple culture media
Enriched culture media
Selective culture media
Differential culture media
a. Simple media
Contain basic nutrients for bacterial growth like
broth with peptone, e. g. Nutrient broth
b. Enriched media
Enriched by some substances like: Blood &
Serum, e.g. Blood agar, Chocolate agar
c. Selective media
Contain substances such as bile salts or antibiotics
that inhibit the growth of some organisms but have
little or no effect on the required organism.
e. g. Salmonella Shigella agar.

d. Differential media
Differential shows up as visible changes,
variations in colony size or in media color, or in
the formation of gas bubbles or precipitates, e.g.
MacConkey agar
Methods used to measure microbial growth
• Count colonies on plate (counts live cells)
• Microscopic counts
• Flow cytometry
• Turbidity
• Count colonies on plate (counts live cells)
• Microscopic counts
• Flow cytometry
• Turbidity
Requirements for Growth
- Bacteria must obtain or synthesize Amino acids,
Carbohydrates & Lipids => build up the cell.
Requirement of growth included:
1. Nutrients
2. Temperature
3. Oxygen
4. pH (potential of hydrogen)
5. Osmotic pressure
- Growth requirements & metabolic by-products
=> Classify different bacteria.
1. Nutrient
-Carbon sources
-Nitrogen sources
-Inorganic salts and trace elements
-Growth factors
-Water
Nutritional types of bacteria

A. Depend on how the organism obtains


carbon for synthesizing cell mass divided
into:
 autotrophic – carbon is obtained from
carbon dioxide (CO2)
 heterotrophic – carbon is obtained from
organic compounds
 mixotrophic – carbon is obtained from
both organic compounds and CO2
B. Depend on how the organism obtains reducing
equivalents used either in energy conservation or
in biosynthetic reactions:
 lithotrophic – red. equiv. are obtained from
inorganic compounds
 organotrophic – red. equiv. are obtained from
organic compounds
C. Depend on how the organism obtains energy for
living and growing:
 chemotrophic – energy is obtained from chemical
compounds
 phototrophic – energy is obtained from light
 chemolithoautotrophs obtain energy from chemical
compounds, red. eque. from inorganic compounds and
carbon from CO2 . e.g.: Knallgas-bacteria
 photolithoautotrophs obtain energy from light,
reducing equivalents from inorganic compounds and
carbon from CO2. e.g.: Cyanobacteria
 chemolithoheterotrophs obtain energy from chemical
compounds and red. eq from inorganic compounds,
carbon by organic compounds . e.g.: Nitrobacter
 chemoorganoheterotrophs obtain energy, carbon,
and reducing equivalents from organic compounds.
e.g.: most bacteria, e. g. Escherichia coli
2.Temperature
 Psychrophiles: cold-loving.
- Best growth berween -15to +15.
- Optimum temperature commonly 0 C.
 Mesophiles: moderate temperature-loving
(Most bacteria)
- Include most pathogens.
- Best growth between 25 to 40 C.
- Optimum temperature commonly 37C.
- Many have adapted to live in the bodies of
human.
Thermophiles: heat-loving

- Optimum growth between 50 to 80 C.


- Many cannot grow below 45 C.
-Adapted to live in sunlit soil and hot springs.
3.Oxygen

(a) Obligate aerobes – require O2


(b) Obligate anaerobes – die in the presence of O2

(c) Facultative anaerobes – can use O2 but also


grow without it

(d) Microaerophilic -requires lower oxygen to


survive.

(e) Aerotolerant anaerobe: tolerate the presence of


oxygen but does not require it for its growth
4. pH
Organisms can be classified as:
 Acidophiles: “Acid loving”.
 Grow at very low pH (0.1 to 5.4)
(many fungi).

 Neutrophiles:
- Grow at pH 5.4 to 8.5.
- Includes most human pathogens.
 Alkaliphiles: “Alkali loving”.
- Grow at alkaline or high pH (7 to 12 or
higher)
- Vibrio cholerae -optimal pH 9.
- Soil bacterium Agrobacterium grows at pH
12.
Most bacteria grow between pH 6.5 and 7.5

Neutrophiles

Alkalophiles
Acidophiles
5. Osmotic Pressure
 Microbes require minerals or nutrients for their
growth, which can be obtained from the
surrounding water.

 Osmotic pressure and salt concentration of the


solution can influence bacterial growth. The
bacterial cell wall gives a mechanical strength that
allows the bacteria to withstand alternations in the
osmotic pressure.
 Osmophilic bacteria requires high osmotic pressure.
When the bacterial cell is subjected to the
hypertonic solution, it may cause osmotic removal
of water, resulting in plasmolysis or osmotic
shrinkage of the protoplasm.

 In contrast, when the bacterial cell is subjected to the


hypotonic solution, it may cause excessive
imbibition of water resulting in plasmoptysis or cell
bursting.

Metabolism
– Sum up all the chemical processes that occur within
a cell

1. Anabolism: Synthesis of more complex


compounds and use of energy

2. Catabolism: Break down a substrate and capture


energy for growth and maintenance.

- All cells require the energy supply to survive. The


common energy form => ATP (Adenosine Tri-
Phosphate)
ATP
is generated by the phosphorylation
of ADP
Microbial metabolism
-Is the means by which a microbe obtains the energy
and nutrients, it needs to living and reproduce.

-Microbes use many different types of metabolic


strategies, and microbes species can often be
differentiated from each other based on metabolic
characteristics.
Metabolism of Glucose
- Bacteria can metabolism of glucose, proteins or
lipids.

-Bacteria can produce energy from glucose. Glucose


breakdown (Glycolysis) can be aerobic (using oxygen)
or anaerobic (without oxygen).

-Anaerobic metabolism of glucose is also known as

anaerobic glycolysis or fermentation.

-Aerobic metabolism of glucose is known as aerobic


glycolysis and respiration.
Catabolism/Aerobic Respiration
of Glucose

The breakdown of carbohydrates to release


energy occur by three pathways:

1. Glycolysis pathway (the Embden-Meyerhof-


Parnas pathway) ---Cytoplasm

2. Krebs cycle --- Cytoplasm (eukaryotes);


cell membrane (prokaryotes).

3. Electron transport chain ---Cytoplasm


Number of ATP Produced From One Molecule of
Glucose by Aerobic Respiration

Biochemical Prokaryotic Eukaryotic


pathways
Glycolysis 2 2

Kreps cycle 2 2

ETC 32 34

Total ATP 36 38
Molecular biology and bacterial genetic
Molecular biology is the study of biology at a
molecular level and understanding the interactions
between DNA, RNA and protein biosynthesis as well
as learning how these interactions are regulated.
The field overlaps with other areas of biology and
chemistry, particularly genetics and biochemistry.
- The Modern of molecular biology begins in the
1930s and the term was first coined by Warren
Weaver in 1938.
- Since the late 1950s and early 1960s, molecular
biologists have learned to Characterize, isolate, and
manipulate the molecular components of cells and
organisms, which are:
1. DNA, the storage of genetic information
2. RNA
3. Proteins, the major structural and enzymatic type of
molecule in cells.
 DNA and RNA
- DNA and RNA are polymers composed of
monomers called nucleotides.
- Each nucleotide has three parts:
a. pentose (5-carbon) sugar.
b. nitrogenous base.
c. phosphate group.
- The pentose sugar in RNA is ribose, and in DNA
it’s deoxyribose.
The only difference is that RNA has a hydroxyl
(OH) group, while DNA has only a hydrogen.
 -There are two classes of nitrogenous bases:
 a. Purines (double-ring) include adenine (A) and
guanine (G).
 b. Pyrimidines (one-ring) include cytosine (C),
thymine (T) in DNA and uracil (U) in RNA.
 Adenine, guanine, cytosine, thymine and uracil
are usually abbreviated using the single letter
codes A, G, C, T and U, respectively.
DNA - Structure
- The structure of DNA was described by Watson and Crick
as long double helix
- Sugar phosphate backbone on the outside and its bases on
inside; the two strand of helix run in opposite direction and
are anti-parallel to each other.
The DNA double helix is stabilized by hydrogen bonds
between the bases
Bacterial genetics
• Bacterial genetics are different from eukaryotic genetics.
One of the major differences between bacterial and
eukaryotic genetics stems from the bacteria's lack of
membrane-bound organelles.
• The genome is all genetic instructions (genes) for
development of cellular structures, metabolic functions, and
their regulation.
• Genes are located within chromosomes (linear or circular) or
plasmids, as specific regions of DNA (or RNA for some
viruses).
Bacterial genetics
• Bacterial genome contains one chromosomal DNA and many
plasmids.
- Like other organisms, bacteria also maintain their
characteristics from generation to generation, yet at the same
time, exhibit variations in particular properties in a small
proportion of their progeny.
Genetic Transfer in Bacteria
• Genetic transfer-results in genetic variation
• Genetic variation-needed for evolution
• Three ways:
• Transformation: genes transferred from one bacterium to
another as “naked” DNA (e.g from lysed cells). New DNA is
incorporated into the chromosome.
• Conjugation: plasmids transferred one bacteria to another
via a pilus.
• Transduction: DNA transferred from one bacteria to
another by a virus.

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