Biological Sciences 2023-2024

Bacterial growth and metabolism

Dr. Avan H. Ghaib

Dept. of Basic Medical Sciences
[email protected]
Learning objectives Upon completion of this lecture, students should be able to:

1) Define Bacterial Growth: Understand the concept of bacterial growth

2) Explain Factors Influencing Bacterial Growth: Identify and explain environmental factors (e.g.,
temperature, pH, nutrient availability) that influence bacterial growth.

3) Describe the Phases of Bacterial Growth: Differentiate between the lag phase, exponential phase,
stationary phase, and death phase in bacterial growth.

4) Describe Key Metabolic Pathways: Explain major bacterial metabolic pathways, such as
glycolysis, the Krebs cycle, and oxidative phosphorylation.

5) Explain Respiration & Fermentation

The physiology of bacteria studies the vital activity of microbial cells – processes of their
nutrition, respiration, growth, and reproduction.
Bacterial growth
In microbiology, growth is defined as an increase in the number of cells rather than in the size
of individual cells. Bacterial growth is asexual reproduction, or cell division, of a bacterium
into two daughter cells, in a process called binary fission. making up a population, referred to
as a culture. Whereas, one cell becomes a colony of millions of cells.
Providing no mutational event occurs the resulting daughter cells are genetically identical to
the original cell. The time required for this process is called the generation time.
Generation time: ” is often used to describe the growth rate
Generation Time: The time required for a cell to divide (and its population to double) Cell’s
division produces two cells, two cells’ divisions produce four cells, and so on. examples’;
The generation time of bacterium in culture can be little as 20 minute for fast growing bacteria
or as long as 24 hour for a slow growing bacteria
-Escherichia coli 20 minutes – -Mycobacterium tuberculosis 18 hours -Mycobacterium
leprae 14 days
 Factors affecting bacterial growth
Growth Factors - Microbes can exist in a great many environments because they are small,
easily dispersed, need only small quantities of nutrients, and are diverse in their nutritional
requirements.
The requirements for microbial growth can be divided into two main categories:
physical and chemical.
▪ Physical aspects include temperature, pH, and osmotic pressure.
▪ Chemical requirements include sources of carbon, nitrogen, sulfur, phosphorus, oxygen,
trace elements, and organic growth factors.
Many factors affect the generation time of the bacterium:

Temperature pH oxygen salt concentration

and nutrients
Most bacteria grow best when these parameters are optimum
A. Physical Factors:
1. pH
The majority of bacteria grow best at neutral or slightly alkaline pH • pH 7.0 –7.4 => this is
near most normal body fluids.
a. acidophiles (acid-loving) – grow best at a pH of 1 to 5.4; Ex One pathogen,
Helicobacter pylori, is adapted to and survives in stomach acid (cause of ulcers)
b. neutrophiles – exist from pH to 5.4 to 7 ; most bacteria that cause human disease are in
this category.
c. alkaliphiles (base loving) – exist from pH to 7.0 to 11.5; ex. Vibrio cholerae (causes
cholera)
2- Temperature
The majority of medically important bacteria are mesophiles, growing at 37°C (i.e body
temperature). However, certain bacteria are capable of growing at extremes of temperature
that would certainly hinder the survival of almost all eukaryotic organisms.

Microorganisms are classified into three primary groups based on their preferred range
of temperature:

▪ psychrophiles (cold-loving microbes), grow between 0°C and 20–30°C Cause food
spoilage

▪ Mesophiles (moderate-temperature-loving microbes), grow best between 25 oC and 40oC;

the human body temp is 37 oC.

▪ Thermophiles (heat-loving microbes). 50 oC to 60 oC; found in compost heaps and in

boiling hot springs
Temprature
According the temperature degree that bacteria survive,
they can classified to :

psychrophiles mesophiles thermophiles extramophiles.

The bacteria that

grow best at ambient The bacteria that have an that grow well at 100- 120 ºC
grow best from
temperatures 20 -40 ºC optimum temperature
about 0 - 20 ºC above 40 ºC
Each bacterial species grows at particular minimum, optimum, and maximum temperatures.

▪ The minimum growth temperature is the lowest temperature at which the species will
grow.

▪ The optimum growth temperature is the temperature at which the species grows best.

▪ The maximum growth temperature is the highest temperature at which growth is possible
Temperature affects growth
Increased temperature breaks chemical bonds
▪ temperature affects bacteria growth by influencing enzymatic activity, cell membrane
properties, protein structure, DNA stability, and metabolic rates(impact of temperature on
various biochemical and physiological processes). Extreme temperatures can lead to the
denaturation and dysfunction of essential biomolecules, ultimately impacting the ability of
bacteria to grow and survive.
3- Osmotic Pressure
Osmotic pressure relative salt concentrations in water solutions
- High osmotic pressure (hypertonic) higher salt concentrations, slows or stops bacterial
growth removes water causing plasmolysis – inhibits growth i.e. salt as preservative.
- Bacteria are more tolerant to osmotic variations because of the mechanical strength of the
cell wall.
some prefer higher salt: Halophiles
some survive and thrive, Vibrio bacteria, Vbiro. cholera
B- chemical Requirements

1. includes many elements like:

A. carbon, hydrogen, O2 , nitrogen, phosphorus & sulphur: needed for the synthesis of
structural components.

B. potassium, calcium magnesium, and iron: needed for cellular functions.

2. Can be obtained from simple elements or by breaking down large molecules such as
protein breakdown into amino acids using bacterial enzymes.
1- Carbon: Besides water, one of the most important requirements for microbial growth is
carbon. Carbon is the structural backbone of living matter; it is needed for all the organic
compounds that make up a living cell. Half the dry weight of a typical bacterial cell is carbon.

Carbon –is necessary for the production of many macromolecules (proteins, lipids, and
carbohydrates)

Nutritional types of bacteria

Depending on how the organism obtains carbon for synthesizing cell mass is divided into:

➢ autotrophic – carbon is obtained from carbon dioxide (CO2) or. An autotroph is an

organism that can produce its food using light, water, carbon dioxide, or other chemicals.

➢ heterotrophic – carbon is obtained from organic compounds. comprise groups of bacteria

that require more complex substances for growth.
2- Nitrogen - needed for amino acids and nucleotides; some can synthesize all 20 amino
acids; others have to have some provided in their medium.

3- Sulfur – needed for amino acids, coenzymes

4-Phosphorus – needed for ATP, phospholipids, and nucleotides

5- Vitamins – a vitamin is an organic substance that an organism requires in small amounts

and that is typically used as a coenzyme; many bacteria make their own, but some are
required in the medium; microbes living in the human intestine manufacture vitamin K,
needed for blood clotting, and some of the B vitamins, thus benefiting their host.

6- Certain trace elements – ex. copper, iron, zinc, sodium chloride, potassium, calcium, etc.;
often serve as cofactors in enzymatic reactions.
7- Oxygen Requirements (for metabolism )
different categories are present in bacteria according to oxygen requirement.
1. strict or obligate anaerobes
▪ grow only in the absence of O2; sensitive to O2 and brief exposure will kill these
organisms; perhaps because these organisms are unable to detoxify some of the products
of O2 metabolism.
▪ lack a respiratory system and can’t use oxygen as a terminal electron acceptor. They
depend on fermentation and anaerobic respiration ex. Clostridium tetani
2. strict or obligate aerobes – lack of oxygen kills the bacteria; ex. Pseudomonas require
molecular oxygen (as a final electron acceptor in catabolism)

3. facultative anaerobes – can shift their metabolism (anaerobic if oxygen is absent or

aerobic if oxygen is present); ex. E. coli, Staphylococcus
4. aerotolerant –Aerotolerant anaerobes use fermentation to produce ATP. the bacteria
don’t use oxygen for growth, but oxygen doesn’t harm them; ex. Lactobacillus.
5. microaerophiles – like low oxygen concentrations and higher carbon dioxide
concentrations; ex. Campylobacter, Streptococci
The bacterial growth curve
Bacterial growth over time can be graphed as cell number versus time. This is called a growth
curve. (dynamics of growth Phases of growth). This curve typically has 4 distinct phases:
(Stages in the Normal Growth Curve)
-Lag phase
-Exponential (Log) phase
-Stationary phase (Post-exponential)
-Decline phase (Death Phase)
Lag phase:
➢ Is the first phase.
➢ It is the period where the individual bacteria are maturing and not yet able to divide(Cells
are actively metabolizing, in preparation for cell division )
➢ During the lag phase synthesis of synthesize proteins and other molecules necessary for
replication. These cells increase in size, but no cell division occurs in the phase.
➢ It may be short or very long, according to the growth medium.
Log Phase
➢ Is the second phase.
➢ The log phase (sometimes called the logarithmic phase or the exponential phase) is a period
characterized by the cells grow most rapidly, doubling at a fairly constant rate.
➢ Number of cells produced > Number of cells dying
➢ Cells are at highest metabolic activity.
➢ Cells are most susceptible to adverse environmental factors at this stage. • Radiation • Antibiotics
➢ Exponential growth cannot continue indefinitely, because the medium is soon depleted of nutrients
and enriched with wastes
Stationary phase

➢ Is third phase .
➢ Population size begins to stabilize.
➢ Number of cells produced = Number of cells dying
➢ Overall cell number does not increase.
➢ Cell division begins to slow down.
➢ Factors that slow down microbial growth:
▪ Accumulation of toxic waste materials
▪ Acidic pH of media
▪ Limited nutrients
▪ Insufficient oxygen supply
➢ The number of cells doesn't increase, but changes in cells occur: cell become smaller and synthesize
components to help them survive longer periods without growing (some may even produce
endospores).
Death or Decline Phase:
➢ Is the final phase .
➢ 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 species, or other injurious conditions
Population size begins to decrease.
➢ Number of cells dying > Number of cells produced
➢ Cell number decreases at a logarithmic rate.
➢ Cells lose their ability to divide.
➢ A few cells may remain alive for a long period of Time
Why study such a growth curve?
▪ Helps us understand how microbes grow under different
conditions
▪ Helps us understand how pathogen grow in our body
▪ Helps us study the effect of different chemicals, osmotic
conditions, even the effect of temperature on bacterial growth.
 Metabolism of Bacteria

Metabolism is a complex of biochemical pathways providing energy accumulation and cell structure synthesis. It
is composed of two closely related sets of reactions: catabolism and anabolism.

Two general types of metabolic reactions

1- Catabolism:

Catabolism is the set of metabolic processes that break down large molecules. Catabolism eventually produces the
chemical energy (ATP) required for all cellular functions such as anabolism (synthesis), membrane transport, etc.

Catabolic Reactions Breakdown of complex substances to small molecules

▪ Degradative reactions

▪ Breaking of covalent bonds

▪ Eg, breakdown of carbohydrates for energy

2- Anabolism:
Synthesis of more complex compounds and use of energy i.e building macromolecules
from small molecules

▪ Biosynthetic reactions(Nutrients present in the environment or growth media must

contain all the elements necessary for the microbial biosynthesis).
▪ Forming of covalent bonds
▪ Requires energy to form bonds
▪ .Eg, making proteins from amino acids
Catabolism and Anabolism
 Energy Generating Process

• Sugars formed or obtained - broken down to release energy

• Aerobic respiration
• Anaerobic respiration
• Fermentation

• Microbes that use aerobic respiration, detoxify generated waste:

Catalase: H2O2 -------→ H20 and 02

Superoxide dismutase : oxygen radical -----→ H20 and O2

Microbes that don’t make these enzymes cannot exist in presence of oxygen
Carbohydrate Catabolism
The breakdown of carbohydrates to release energy (in the form of ATP)
Carbohydrates are catabolized by: Cellular respiration and Fermentation :

1- Cellular respiration : (C6H12O6 + 6O2 + 38ADP+ 38P → 6CO2 + 6H2O +38ATP)

Respiration is a type of heterotrophic metabolism that uses oxygen and in which 38 moles of ATP are
derived from the oxidation of 1 mole of glucose, yielding 380,000 cal. Glucose is the most common
substrate used for studying heterotrophic metabolism. Most aerobic organisms oxidize glucose
completely to CO2 and H2O.
The complete oxidation of glucose may involve three fundamental biochemical pathways
➢ Utilizes glycolysis
➢ Krebs cycle,
➢ Electron Transport and Oxidative Phosphorylation;
Types of respiration :
➢ Aerobic respiration: final electron acceptor is O₂ and occurs in aerobes.
➢ Anaerobic respiration: final electron acceptor is inorganic molecule other than O₂

2- Fermentation
 Cellular respiration pathways
1- Glycolysis
➢ -- A nine-step biochemical reaction, each of which requires specific enzymes. Six-carbon
molecule of glucose is broken down into three-carbon molecules of pyruvic acid
(Oxidation of glucose to pyruvic acid)
➢ - Produces very little energy–only 2 ATP
➢ Takes place in the Cytoplasm of both prokaryotic and eukaryotic cells.
➢ Glycolysis is an oxygen-independent metabolic pathway, meaning that it does not use
molecular oxygen (i.e. atmospheric oxygen) for any of its reactions.

➢ The energy released in the pathway is used to produce two types of energy-rich
molecules:
•Two molecules of ADP are phosphorylated to ATP.
•Two molecules of NAD+ are reduced to NADH/H+.

Result
- Glucose ----->2 Pyruvate + 2ATP + 2NADH
 Cellular respiration
2- Krebs Cycle –
o The Krebs cycle is named after its discoverer, Hans Krebs. It is also known as the citric acid
cycle or the tricarboxylic acid cycle.
o It is a series of chemical reactions required for cellular respiration; it involves redox,
dehydration, hydration, and decarboxylation
o Pyruvate formed at the end of glycolysis is first oxidized into Acetyl CoA which then enters
the citric acid cycle.
o The overall reaction/ equation of the citric acid cycle is:
(Oxidative Decarboxylation of pyruvate to Acetyl CoA)
Acetyl CoA + 3 NAD+ + 1 FAD + 1 ADP + 1 Pi → 2 CO2 + 3 NADH + 3 H+ + 1 FADH2 + 1 ATP
 Cellular respiration
3- Electron Transport and Oxidative Phosphorylation
o Certain of the products produced during the Krebs cycle enter the electron transport
chain
o Consists of a series of oxidation-reduction reactions, whereby energy is released as
electrons are transferred from one compound to another.
o Oxygen is the end of the chain; referred to as the final or terminal electron acceptor.
o Cytochrome oxidase enzyme responsible for transferring electrons to oxygen.
o The enzymes involved in electron transport and oxidative phosphorylation reside on the
bacterial inner (cytoplasmic) membrane. This membrane is invaginated to form structures
called respiratory vesicles, lamellar vesicles, or mesosomes, which function as the
bacterial equivalent of the eukaryotic mitochondrial membrane.
o Produces 34 ATP in prokaryotic cells, and 32 ATP in eukaryotic cells
❖ In addition, two other glucose-catabolizing pathways are found in bacteria: the oxidative
pentose phosphate pathway (hexose monophosphate shunt), and the Entner-Doudoroff
pathway, which is almost exclusively found in obligate aerobic bacteria.
Fermentation
o Scientific definition: is an anaerobic process performed by a cell to generate chemical
energy (e.g. ATP) from pyruvate (a product of glycolysis) but without going through the
citric acid cycle and the electron transport chain system as cellular respiration does.
o In fermentation, another type of heterotrophic metabolism, an organic compound rather
than oxygen is the terminal electron (or hydrogen) acceptor. Less energy is generated
from this incomplete form of glucose oxidation, but the process supports anaerobic
growth.
o Does not require oxygen
o These organic end products are secreted into the medium as waste metabolites (usually
alcohol or acid).
How does bacterial fermentation affect humans?

Lactic acid production. The resulting low pH discourages growth of competing microbes.
Lactobacilli live in very high numbers in the vaginal tract and in the intestine.
Lactobacillus spp. are commensal organisms. Their production of lactic acid creates a
low pH environment that is very important for health in places including the vagina.
Lactobacilli are thought to be particularly important for protecting against Candida
albicans (yeast infections). Some species of lactic acid-producing bacteria are important
in fermenting foods, such as yogurt, and certain cheeses
 Lipid Catabolism

LIPID =
•1 glycerol
•2 fatty acids
Protein Catabolism

▪ Protein is too large to pass through plasma membrane

▪ Extracellular enzymes break down protein into amino acids that can pass through plasma
membrane

▪ Amino acids are enzymatically converted to organic acids that can enter the Krebs
cycle
Deamination, decarboxylation,
dehydrogenation
Organic acid Krebs cycle
• References:
1- Jawetz Melnick & Adelbergs Medical Microbiology 27 E 27th Edition

by Karen Carroll (Author), Janet Butel (Author), Stephen Morse (Author)

2- Murray's Basic Medical Microbiology, 2nd Edition

3-Textbook of 2nd Edition Subhash Chandra Parija MBBS, MD, PhD, DSc, FRCPath
FAMS, FICPath, FICAI, FABMS, FISCD, FIAVP, FIATP, FIMSA Professor and Head
Department of Microbiology Jawaharlal Institute of Postgraduate Medical Education and
Research Puducherry, India

4-Microbiology. Lansing M Prescott, John P. Harley, Donald A. Klein.2004. Sixth

Edition. Higher Education.