CHE 366

INDUSTRIAL MICROBIOLGY

WEEK 3
MICROBIAL GROWTH and NUTRITION

Dr. Burcu ÖZTÜRK

Food Engineering Department
Office No: B116/A
[email protected]
 MICROBIAL NUTRITION

Primary ingredients required by all living organisms include: Microorganisms are made of :
• CHO’s,
• lipids,
• carbon source, • proteins,
• nucleic acids
• water,

• minerals,

• nitrogen source

B.ÖZTÜRK 2
• To obtain energy and construct new cellular components, organisms must have a supply of raw material
or nutrients.

• Nutrients are required for anabolic and catabolic processes of the cell and are required for growth.

Function of the nutrients :

✓ Generation of energy (catabolic reactions)

✓ Synthesis of cellular materials ( anabolic reactions)

B.ÖZTÜRK 3
Metabolism : the Greek metabole, meaning change.

✓ the sum of the biochemical reactions required for energy generation AND (catabolism)

the use of energy to synthesize cell material from small molecules in the environment (anabolism).

environmental conditions: pH, moisture, temperature, salinity

B.ÖZTÜRK 4
 MICROBIAL NUTRITION
MICROBIAL METABOLISM

Why do we must know the metabolism of microorganisms?

✓Because we want to know how to inhibit or stop bacterial growth and want to control their metabolism.
✓The microbial physiology of microorganisms are common to many living systems; since carbohydrates,
fatty acids, amino acids, nitrogeneous bases occur in all cells.
Sources of energy for all living cells (biological systems) :
1- Chemical sources
2-Sunlight

➢ A cell is a miniature factory

➢ A large number of chemical reactions are occurring

➢ A (reactants) + B (reactants) ----> C (products)

➢ Chemical reactions are either endogonic (req. energy) or exogonic (release energy)

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MICROBIAL NUTRITION
METABOLIC DIVERSITY

Cellular metabolism is classified into nutritional groups on the basis of three major criteria:

1.Source of energy, used for growth

2. Source of carbon, and
3. Source of electron donors used for growth.

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MICROBIAL NUTRITION
METABOLIC DIVERSITY NUTRITIONAL TYPES OF MICROORGANISMS

1. ENERGY SOURCE
a. Phototrophs —can use light energy

b. Chemotrophs —must obtain energy from oxidation-reduction of external organic/inorganic chemical

compounds

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

3. ELECTRON SOURCE
a. Lithotrophs- can draw electrons from inorganic molecules

b. Organotrophs- can draw electrons/hydrogens from organic molecules

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 MICROBIAL NUTRITION
METABOLIC DIVERSITY

These requirements can be combined

Major nutritional types are determined by grouping them in the following sequence of sources :

Major Nutritional Types Representative organism

Photolithotrophic autotrophy Algea, cyanobacteria

(photoautotrophs)

Photoorganotrophic heterotrophy Purple non sulfur bacteria

(photoheterotrophs)

Chemolithotrophic autotrophy Nitrifying bacteria

(chemoautotrophs)

Chemoorganotrophic heterotrophy Protozoa, fungi, most non-photosynthetic

(chemoheterotrophs) bacteria, animals

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B.ÖZTÜRK 9
• Over 90 percent of the elemental analysis of cells consists of carbon,

hydrogen, oxygen, nitrogen, phosphorus and sulfur.

(C, H, O, N, P and S are the constituents of organic material).

• These are the elements that become combined to form all the

biochemicals and macromolecules that comprise living systems.

• H and O are the constituents of water (H2O), that makes up over 95

percent of the cell composition.

• Calcium (Ca++), iron (Fe++), magnesium (Mg++) and potassium (K+) are

present as inorganic salts in the cytoplasm of cells.

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NUTRIENTS

Microbial nutrition are classified into two types based on the requirements
2 types of nutrients
1. Macronutrients – needed in large quantities for cellular metabolism & basic cell structure
➢ Major elements : C, H, O, N, S, P (g/l culture medium)
➢ Minor elements : K, Ca, Mg, Fe (mg/l culture medium)

2. Micronutrients – needed in small quantities; more specialized (enzyme & pigment structure &
function)
➢ Trace elements : Mn, Zn, Co, Ni, Cu (чg/l culture medium)

Growth factors – required in very small amounts, can not be synthesized by some cells.
➢ These are vitamins and organic molecules.
Fastidious Bacteria: microbes that require other complex - nutrients/growth factors ( i.e.,
Vitamins or Amino Acids)
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• Microorganisms need macro elements (C, H &O) and electron sources.

• Carbon, Hydrogen & Oxygen molecules are components of organic molecules.

• Electron sources are used for other cellular works and also drive redox reactions.

• Nitrogen is involved in the biosynthesis of the amino acids, nitrogenous bases i.e., purines &
pyrimidines, etc.

• Phosphorous is there in phospholipids, nucleic acids.

• Sulfur is necessary for the biosynthesis of amino acids like cysteine & methionine, and other
cofactors such as thiamine & biotin.

B.ÖZTÜRK 12
MICROBIAL NUTRITION
Major elements, their sources and functions in cells
Element % of dry Source Function
weight

Carbon 50 organic compounds or CO2 Main constituent of cellular material

Oxygen 20 H2O, organic compounds, CO2, and Constituent of cell material and cell water; O2 is electron
O2 acceptor in aerobic respiration
Nitrogen 14 NH3, NO3, organic compounds, N2 Constituent of amino acids, nucleic acids nucleotides, and
coenzymes
Hydrogen 8 H2O, organic compounds, H2 Main constituent of organic compounds and cell water

Phosphorus 3 inorganic phosphates (PO4) Constituent of nucleic acids, nucleotides, phospholipids, LPS,
teichoic acids
Sulfur 1 SO4, H2S, S, organic sulfur Constituent of cysteine, methionine, glutathione, several
compounds coenzymes
Potassium 1 Potassium salts Main cellular inorganic cation and cofactor for certain enzymes

Magnesium 0.5 Magnesium salts Inorganic cellular cation, cofactor for certain enzymatic reactions

Calcium 0.5 Calcium salts Inorganic cellular cation, cofactor for certain enzymes and a
component of endospores
Iron 0.2 Iron salts Component of cytochromes and certain nonheme iron-proteins and
a cofactor for some enzymatic reactions
B.ÖZTÜRK 13
NUTRIENTS/ GROWTH FACTORS
• Some cells require certain organic compounds in minute quantities – Growth Factors.

• They are needed for coenzymes or functional groups of certain enzymes and protein synthesis.

• Most of microorganisms can synthesize these growth factors, but some require them from the culture
medium.

• The need for a growth factor results from either

- a blocked metabolic pathway or

- a missing metabolic pathway

• Auxotroph : a mutated microorganism that lacks the ability to synthesize an essential nutrient and
therefore must obtain it from its environment.

Auxotrophs require growth factors that must be included in their cultivation medium for survival.

• Prototroph : a microorganism requiring the same nutrients as most members of its species. These type
of m.orgs grow on minimal media.

A Prototroph may mutate and change to an Auxotrophic

B.ÖZTÜRK microorganism. 14
NUTRIENT UPTAKE
Movement through cell membrane

• Passive Transport
• Simple diffusion
• diffusion of nonpolar, hydrophobic molecules
• lipids
• high → low concentration gradient
• Facilitated transport
• diffusion of polar, hydrophilic molecules
• through a protein channel
• high → low concentration gradient
• Active transport
• diffusion against concentration gradient
• low → high
• uses a protein pump
• requires ATP ATP

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NUTRIENT UPTAKE

Utilization of High Molecular Weight Materials

• Utilization of polymeric substrates • For organisms with a rigid cell wall this
(polysaccharides, proteins and lipids) requires is not usually possible.

additional activities. • They must secrete extracellular

hydrolytic enzymes—proteases,
• Protozoa and other eukaryotic organisms without
amylases, cellulases, etc.—directly
cell walls can ingest relatively large pieces of food
into the environment and then take up
materials from their environment by phagocytosis
the resultant hydrolysis products.
(engulfment) into a membrane-bound food
• Many of these extracellular enzymes
vacuole.
have major industrial applications
• Hydrolytic enzymes are then secreted into the

vacuole to break down the polymers to their

constituent monomers. B.ÖZTÜRK 16

NUTRIENT UPTAKE

Ionophores
An ionophore is a lipid-soluble molecule usually
synthesized by microorganisms to
transport ions across the lipid bilayer of the cell
membrane.

The two broad classifications of ionophores are:

1)Chemical compounds (mobile ion carriers) that bind

to a particular ion, shielding its charge from the
surrounding environment, and thus facilitating its
crossing of the hydrophobic interior of the lipid
membrane

2)Channel formers that introduce a hydrophilic pore

into the membrane, allowing ions to pass through
while avoiding contact with the
membrane's hydrophobic interior.
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• Ionophores are compounds that can cause leakage of ions across the cell
membrane by destroying the selective permeability properties of
membranes.

• They are hydrophobic; dissolve in the lipid bilayer; allow passive

diffusion of ions into and out of the cell.

• Antibiotics such as valinomycin and tyrocidin allow massive leakage of

ions from certain cells and thus have a lethal effect.

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 What is the Difference btw Ion channels and Ionophores?

• Ion channels and ionophores are both involved in the transport of ions across cell
membranes, but they differ in their mechanisms and functions.
• Ionophores also facilitate the movement of ions across the membrane, and also down the
electrochemical gradient.
• But unlike ion channels, they do not form channels.
• Instead, the facilitate by being lipid soluble, and being able to shield the charge of the ion(s)
in question from the lipid

• Ion channels are protein channels that selectively transport ions across cell membranes,
while ionophores are small molecules that can bind to and transport ions across cell
membranes, but are not selective.

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NUTRIENT UPTAKE

Concentration of water
• Direction of osmosis is determined by comparing total solute concentrations
• Hypertonic - more solute, less water
• Hypotonic - less solute, more water
• Isotonic - equal solute, equal water

• Cell survival depends on balancing water

uptake & loss

hypotonic hypertonic
net movement of water
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• Water is essential for cell metabolsim and growth.
• Osmosis: Passive diffusion of water accross semipermeable membrane.
• Cells are 80 to 90% water and microbial growth is best when osmotic pressure
ideal.
• Normally, salt concentration of microbial cytoplasm ~ 1%.
• Isotonic Solution: When external environment also has a 1% salt concentration,
the osmotic pressure is optimum.

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• Hypertonic solutions: High osmotic pressure removes water from cell, causing
shrinkage of cell membrane (plasmolysis).
• Used to control spoilage and microbial growth.
• Sugar in jelly.

• Salt on meat.

• When the external salt concentration rise, such as food is salted, water will
flow out of microbial cytoplasm by osmosis through cell membrane into
environment, thereby causing microorganisms to shrink and die.

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• Hypotonic solutions: Low osmotic pressure causes water to enter the cell.
• In most cases cell wall prevents excessive entry of water.
• Microbe may lyse or burst if cell wall is weak.
• If exterior water is free of salt, it will flow through cell membrane into
cytoplasm of cell, causing organism to swell and burst.

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B.ÖZTÜRK 24
Microbial Growth
• Microbial Growth: refers to an increase in cell number, not in cell size.
• A characteristic of microorganisms is their ability to grow and form a population of organisms.

• Many microbes are unicellular, meaning they are made of only one cell.

• The size of any unicellular microbe is limited by the capacity for the essential components of
the cell to support its survival.

• Many requirements for successful growth include those both Chemical and Physical.

• Bacteria grow and divide by Binary Fission, a rapid and relatively simple process.

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 Physical Requirements for Growth
• Microbes have a variety of physical requirements for growth, including temperature, pH,
and water stress.
1. Temperature:
Most microbes grow optimally within a certain temperature range dictated by
the ability of proteins within the cell to function.
• Temperature at which best growth occurs is Optimum Growth
Temperature.
• In general, at low temperatures, microbes grow slower. At optimum
temperatures, microbes grow more quickly.
• For instance, pathogens often grow best at normal body temperature, but
slowly at cooler temperatures outside the body or when body
temperature increases during a fever.
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• Each and every m.org. has a cardinal range of temperature for its growth

• The temp. below which growth of m.org. does not take place is called as

Minimum growth temp

• The temp. at which there is an max. growth of m.org. is called as

Optimum growth temp.

• The temp. Above which growth of m.org. Does not take place is called as

Maximum growth temp.

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Physical requirements

Microorganisms classified into groups

according to their temperature
preferences:

• Psychrophilic organisms (Psychrophiles)

prefer cold temperatures of about 0°C
to 20°C

• Mesophilic organisms (Mesophiles)

prefer temperatures at 20°C to 40°C

• Thermophilic organisms (Thermophiles)

prefer temperatures higher than 40°C

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Physical requirements

Psychrophiles (cold-loving)

• Psychrophiles are microbes that can grow at 0°C. Some are inhibited at higher temperatures,
preferring to live in cold climates, whereas others can survive in conditions above 20°C.

• The range for Psychropiles is –10°C to 20°C, with an optimum at about 10°C. They are
sensitive to temperatures over 20°C. Found in very cold environments such as ocean depths,
Arctic, and the Antarctic regions. They seldom cause disease or food spoilage.

• Ex: Arthrobacter sp., Psychrobacter sp.

• Since psychrophiles are unable to survive at a temperature higher than 20 °C, they may
be killed by exposure to room temperature.

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• Psychrotrophs (also known as Psychrotolerant) prefer colder
temperatures but can live just fine in higher ones.

• The range for Psychrotrophs is from 0°C to 40°C with an optimum of

about 20°C. They are found in many natural environments in temperate
climates and are responsible for the spoilage of refrigerated food.

• Ex: Listeria monocytogenes

• They are more likely to be found in environments that are seasonally

cold. Psychrotrophs are more abundant in nature than psychrophiles.

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Mesophiles (middle-loving)
• Mesophiles like it best between 25°C and 40°C but can survive between 10°C and 50°C.

• Microbes that live within animals grow optimally at a temperature that matches that of
their host. For instance, microbes that live in the human body grow between 34 and 37°C,
which is body temperature.

• Most of the pathogenic microorganisms and normal human microbiota are mesophiles!!!

• E.g. Staphylococcus aureus, Escherichia coli

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Thermophiles (Heat-loving)

• Thermophiles can tolerate temperatures up to 70°C and like it best between

50°C and 60°C.

• Thermophiles are widely distributed in hot springs, geothermal soils, sunlit soil and manmade
environments such as garden compost piles where the microbes break down organic waste.

• They possess a significant potential for the degradation of environmental pollutants.

• Thermophilic and thermotolerant microorganisms are of important economic value due to their
ability to produce thermostable extracellular enzymes which have important biotechnological
applications.

• Ex: Thermus aquaticus and Geobacillus spp

• Some thermophiles form extremely heat resistant endospores.

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• This group also contains a subset considered Hyperthermophilic, or extreme heat loving.

• All the known microorganisms in this category are Archaea and some can even grow in
temperatures above 120°C, deep in the sea where the pressure stops water from boiling
at that temperature.

• Their enzymes can serve as model systems for use by biologists, chemists, and
physicists interested in understanding enzyme evolution, molecular mechanisms for
protein thermostability, and the upper temperature limit for enzyme function.

• Archaebacteria. Most live in volcanic and ocean vents

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2. pH

• The pH is the measure of how acidic or alkaline a solution is, with

values from 0 to 14.

• Neutral pH is around 7. Alkaline, or basic pH, is 8 to 14.

• Most bacteria prefer neutral pH (6.5 to 7.5), whereas Fungi can

grow in more acidic conditions, preferring pH 5 to 6.

• Some bacteria and Archaea are acididophilic and they grow in

conditions far too acidic for other species.

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• Alkaliphiles: (Alkali loving) Grow at alkaline (pH 9) or high pH
(7 to 12 or higher)

• Vibrio cholerae and Alkaligenes faecalis optimal pH 9.

• Soil bacterium Agrobacterium grows at pH 12.

• Neutrophiles: Grow at pH 5.4 to 8.5.

• Since pH of most human tissue is 7.0 to 7.2, these

Neutrophilic bacteria usually grow well in body.

• Includes most human pathogens

• Acidophiles (Acid loving) Grow at very low pH (0.1 to 5.4)

• Lactobacillus produces lactic acid, tolerates mild acidity.

• Molds & yeasts common acidophilic microorganisms

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3. Osmotic pressure
• Water is essential for cell metabolsim and growth.
• Osmosis: Passive diffusion of water accross semipermeable membrane.
• Cells are 80 to 90% water and microbial growth is best when osmotic
pressure ideal.
• Normally, salt concentration of microbial cytoplasm ~ 1%.
• Isotonic Solution: When external environment also has a 1% salt
concentration, the osmotic pressure is optimum.

B.ÖZTÜRK 36
• According to the osmotic pressure;

• Halophiles: Require moderate to large salt concentrations.

Ocean water contains 3.5% salt.
• Most bacteria in oceans.
• Extreme or Obligate Halophiles: Require very high salt
concentrations (20 to 30%).
• Ex: Halobacterium salinarum . Bacteria in Dead Sea, brine vats.
• Facultative Halophiles: Do not require high salt
concentrations for growth, but tolerate 2% salt or more.
• Ex: Staphylococcus epidermis (skin )

B.ÖZTÜRK 37
 Chemical Requirements for Growth

• Unlike the physical requirements where a specific range or

concentration is necessary for optimum growth

• The chemical requirements just need to be present in the

environment and a microbe will use what it needs.

• Microbes use compounds , elements and vitamins to make

everything in the cell including membranes, proteins, and
nucleic acids.

B.ÖZTÜRK 38
 Chemical requirements

1. Carbon:
➢ Carbon is one of the chemical elements found in nature and it is
necessary for all life.
• It can be obtained from organic materials in environment, or it may
be derived from carbon dioxide.
• Perhaps 50 % of a bacterium's dry weight is carbon.
• Structural backbone of all organic compounds.

• Chemoheterotrophs: Obtain carbon from their energy source:

lipids, proteins, and carbohydrates.

• Chemoautotrophs and Photoautotrophs: synthesize all necessary

organic compounds from carbon dioxide.
• They derive energy sources from chemical rxn.

B.ÖZTÜRK 39
2. Nitrogen, sulfur, and phosphorus:

• They are necessary for protein and nucleic acid biosynthesis.

• Phosphorus is essential to biological information storage and transfer,

energy metabolism, membrane integrity and construction of
phospholipids.
• Many bacteria can reduce sulfur in small amounts, but some specialized
bacteria can perform respiration entirely using sulfur. They use sulfur or
sulfate as an electron receptor in their respiration, and release sulfide
as waste. This is a common form of anaerobic respiration in microbes.
• Nitrogen is used for the synthesis of proteins, amino acids, DNA, and
RNA.
• Most microbes get these elements by degrading proteins and nucleic acids, but some
capture nitrogen from nitrogen gas or ammonia or get sulfur from other ions in the
environment.

B.ÖZTÜRK 40
3. Trace elements:

Trace elements such as iron, copper, molybdenum, and zinc are needed as
cofactors for enzymes and must be obtained, in tiny amounts, from the
environment.

4. Vitamins and Amino acids:

Microbes can make vitamins, which also act as enzyme cofactors.

• Some microbes, however, lack the ability to make one or several vitamins and
have to get them from their environment.

• The same is true of amino acids and these along with the vitamins needed
are called Growth Factors.

• Although most bacteria can make all the amino acids they need, some can’t
quite make them all; these bacteria are called Auxotrophs.

B.ÖZTÜRK 41
5. Oxygen :

The presence of oxygen affects a few different aspects of microbial growth .

• Organisms that use molecular oxygen (O2), produce more energy from nutrients than
anaerobes.

• Oxygen is used by aerobic bacteria during process of cellular respiration as a final

electron acceptor.

• Different microbes respond differently to oxygen.

• For Aerobic organisms, oxygen is an absolute requirement for their energy-yielding

properties.

• Certain microorganisms grow in oxygen-free environments and are described as

Anaerobic.
B.ÖZTÜRK 42
• The term Obligate refers to the absolute need for something, whereas the term
facultative means that they can function in conditions that are not their preferred
conditions.

➢ Microorganism can classify based on their oxygen requirements:

• Obligate Aerobes: Require oxygen to live.
• They grow well at full oxygen levels (about 21 percent in air).

• Disadvantage : Oxygen dissolves poorly in water.

• Example: Pseudomonas, common nosocomial pathogen

B.ÖZTÜRK 43
P. aeruginosa
• Facultative Anaerobes: Can use oxygen, but can grow in its absence. Have complex set of
enzymes.
• Examples: E. coli, Staphylococcus, yeasts, and many intestinal bacteria.

Staphylococcus aureus
• Obligate Anaerobes: They are either inhibited or killed by oxygen. They live in
environments that are devoid of all oxygen, like aquatic sediments or the colon of animals.
• Examples: Clostridium bacteria that cause tetanus and botulism.

Clostridium botulinum
B.ÖZTÜRK 44
• Aerotolerant Anaerobes: Can’t use oxygen, but tolerate its presence.
Can break down toxic forms of oxygen.

• Example: Lactobacillus
• They ignore oxygen in their environment and can grow well in it’s presence or absence.
Because they lack the citric acid cycle and/or an electron transport chain, they don’t
switch to aerobic respiration when oxygen is present.

Lactobacillus sp.
• Microaerophiles: Microaerophiles prefer to grow in conditions where the concentration of
oxygen is very low. Sensitive to toxic forms of oxygen.
• Example: Campylobacter.

Campylobacter jejuni B.ÖZTÜRK 45

Microbial Growth Kinetics
Microbial growth : an orderly increase in cellular components, resulting in cell enlargement and
eventually leading to cell division.

This definition is not strictly accurate as it implies that a consequence of growth is always an
increase in cell numbers.

However, under certain conditions growth can occur without cell division, for example, when cells are
synthesizing storage compounds, e.g. glycogen or poly b-hydroxybutyrate.

• In this situation the cell numbers remain constant, but the concentration of biomass continues to
increase.

This is also true for coenocytic organisms, such as some fungi, that are not divided into separate cells.

• Their growth results only in increased size.

B.ÖZTÜRK 46
• The growth model that will be examined is bacterial binary fission in homogeneous
suspension cultures, where cell division produces identical daughter cells.

• Each time a cell divides is called a generation and the time taken for the cell to divide is referred
to as the generation time.

• Therefore, the generation time or doubling time (td) is the time required for a microbial
population to double.

• Theoretically, after one generation, both the microbial cell population and biomass
concentration have doubled.

B.ÖZTÜRK 47
if we start from one bacteria, it divides after every generation time as follows

After n generations, no of bacteria will be

Eq 1

Eq 2

Eq 3

Eq 4

Eq 2 can be used to determine number of bacteria, if initial number of bacteria and number of
generation is known where as Eq 4 can directly been used to calculate number of generations.

B.ÖZTÜRK 48
Microbial fermentations in liquid media can be carried out under different operating conditions, i.e. batch growth,
fed-batch growth or continuous growth.

Batch growth involves a closed system where all nutrients are present at the start of the fermentation within a
fixed volume. The only further additions may be acids or bases for pH control, or gases (e.g. aeration, if
required).

In fed-batch systems fresh medium or medium components are fed continuously, intermittently or are added as a
single supplement and the volume of the batch increases with time.

Continuous fermentations are open systems where fresh medium is continuously fed into the fermentation vessel,
but the volume remains constant as spent medium and cells are removed at the same rate.

B.ÖZTÜRK 49
 Microbial Growth in Batch Culture

decelaration

acceleration

Trophophase Idiophase
(primary metabolism) (secondary metabolism)
Only observed in some microorganisms

http://www.youtube.com/watch?v=Q9OQkbq7vKw

B.ÖZTÜRK 50
 Batch Growth
Batch Growth
Lag phase

A period of adaptation for the cells to their new environment

• New enzymes are synthesized.

• A slight increase in cell mass and volume, but no increase in

cell number

• Prolonged by low inoculum volume, poor inoculum condition

(high % of dead cells), age of inoculum, nutrient-poor medium.

• Multiple lag phases: (diauxic growth) medium contains more

than one carbon source

B.ÖZTÜRK 51
Batch Growth
Exponential Growth Phase

Once the cells have adapted to their new environment they enter the acceleration
phase.

Cell division occurs with increasing frequency until the maximum growth rate (μ max)
for the specific conditions of the batch fermentation is reached.

At this point exponential growth begins and cell numbers/biomass increase at a

constant rate.

Mathematically, this exponential growth can be described by two methods;

1) related to biomass (x) and
2) related to cell numbers (N).

B.ÖZTÜRK 52
 Batch Growth
Exponential Growth Phase

For cell biomass, growth can be considered as an autocatalytic reaction.

Therefore, the rate of growth is dependent on the biomass concentration, i.e.

catalyst, that is present at any given time.

This can be described as follows:

rate of change of biomass is dx/dt =μx where
x = concentration of biomass (g/L),
μ = specific growth rate (per hour)
t = time (h).

B.ÖZTÜRK 53
Batch Growth

Deceleration growth phase

Very short phase, during which growth decelerates due to

either:

• Depletion of one or more essential nutrients

• The accumulation of toxic by-products of growth (e.g. Ethanol in yeast

fermentations)

• Period of unbalanced growth: Cells undergo internal restructuring to increase their

chances of survival

B.ÖZTÜRK 54
Batch Growth

Stationary Phase

With the exhaustion of nutrients and build-up of waste and

secondary metabolic products :
- The growth rate equals the death rate.

- There is no net growth in the organism population.

- Cells may have active metabolism to produce secondary metabolites.

Primary metabolites are growth-related: ethanol by S. cerevisae.

Secondary metabolites are non-growth-related: antibiotics, pigments.

B.ÖZTÜRK 55
Batch Growth
Death Phase:
The living organism population decreases with time, due to a lack
of nutrients and toxic metabolic by-products.

The rate of death usually follows:

dN
= −kd N
'

dt
'
k d is the first - order death rate constant.

B.ÖZTÜRK 56
APPLICATION OF BATCH FERMENTATIONS

• During batch fermentations certain environmental conditions continually

change, particularly nutrient and product concentrations, as does the
specific growth rate, because the cells must pass through the sequence of
growth phases described before.

• Consequently, the system never achieves steady-state conditions.

• A further disadvantage is that several distinct practical stages are

associated with the operation of a batch fermentation:

B.ÖZTÜRK 57
Operational stages of a batch fermentor/growth/process
1. charging of the fermenter with fresh medium;
2. sterilization of the fermenter and medium;
3. inoculation of the fermenter;

4. production of microbial products;

5. harvesting of biomass and spent fermentation broth;
and finally
6. cleaning of the vessel.

This has major economic implications for industrial processes.

For a considerable period of time, the fermentation vessel is not producing microbial
products, but is being cleaned, filled, sterilized, etc.

The non-productive period is referred to as the downtime of the fermenter.

B.ÖZTÜRK 58
Batch Growth

B.ÖZTÜRK 59
 Growth Kinetics

• Cell growth and cell division are inseparable for microbes as

bacteria divide by binary fission, yeast cells by budding
• Growth kinetics is an autocatalytic reaction which implies that
the rate of growth is directly proportional to the concentration
of cell.
• Microbial growth kinetics, the relationship between the specific
growth rate (μ) of a microbial population and the substrate
concentration (s)
• Mathematical models are kinetic models which explain the
relationship between rates and the concentration of
reactants/products and allows to predict the rate of conversion
of reactions in to products.

B.ÖZTÜRK 60
Growth kinetics
Classified based on the relationship btw product synthesis and energy generation in
the cell:
•Growth associated
•Non-growth associated
•Mixed-growth associated

Primary
metabolite Secondary
metabolite

Growth-associated Mixed-growth-associated Non growth-associated

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1. Growth associated
• Growth linked products are formed by growing cells and hence primary
metabolites.
• That is product concentration increases with cell concentration.
• The formation of growth associated product may be described by Eq1.

dP/dt=qp X Eq1.

where P = concentration of product

qp = specific rate of product formation
X = biomass concentration.

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• The product synthetic and energy generation have created these
growth-dependent products.
• Growth-linked products have been created through the growing cells.
• Several elements have influenced the microbial growth of the
products.
• Nutrients, temperature, Moisture, pH levels have stimulated the
growth of the predictions in the product kinetics.

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2. Non-growth associated
• They are formed by cells which are not metabolically active and hence
are called secondary metabolites.
• The formation of Non-growth associated product may be described
by Eq.2

qp = ꞵ constant Eq.2

Ex: ethanol has been the process of non-

growth associated product formation.
Ethanol has been an extracellular product. Non-growth associated products are the
function of the concentration of the cells.
B.ÖZTÜRK
• These secondary metabolites from non-growth products have been
synthesized through the stationary phase of the microbial culture.
• These secondary metabolites have supported the short phase of
exponential which extended the production phase.
• This exponential phase has been maintained and controlled through
the addition of the fresh and medical to the vessels as it moves the
cultured median from a certain period.

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• Mixed growth associated products- products are formed during the
slow growth and stationary phases.
• Produced during deceleration and stationary phases
Example: secondary metabolites, production of lactic acid and
xanthan gum

qp = αμ +ꞵ Eq. 3

• Products are formed during the slow growth and

stationary phases.
• Produced during deceleration and stationary
phases
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Products
• Growth-associated
• produced at the same time as cell growth
• constitutive enzymes (ones that are normally present)
• glucose isomerase
• metabolic intermediates
• pyruvate, citrate, acetate
• Non-growth-associated
• takes place during the stationary phase (m=0)
• secondary metabolites
• antibiotics
• Mixed - growth associated
• takes place during growth and stationary phases
• metabolic byproducts
• lactate, ethanol
• secondary metabolites

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 Production kinetics
• Microbial growth kinetics, the relationship between the specific growth
rate (μ) of a microbial population and the substrate concentration (s)
• Monod models is used to describe the batch growth kinetics of cell.
• The Monod kinetic model is given as Eq. 4

Eq. 4
where μ is the specific growth rate (h-1),
S is substrate concentration (g/L) and
KS is the Monod constant (g/L)
and μmaxmaximum specific growth rate, (h-1)
The yield coefficient and the specific growth rate
used to develop three types of microbial growth
kinetic relationships; Monod , first order ,and zero
order kinetics.

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