MICROBIAL REQUIREMENTS FOR GROWTH

• Physical requirements
– Temperature
– pH
– Osmotic pressure

• Chemical requirements
– Carbon
– Nitrogen, sulfur, and phosphorous
– Trace elements
– Oxygen
– Organic growth factor
 Temperature and Microbial Growth
• Cardinal temperatures
– minimum
– optimum
– maximum
• Temperature is a major
environmental factor
controlling microbial
growth.

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 Temperature
• Minimum Temperature: Temperature below which
growth ceases, or lowest temperature at which
microbes will grow.

• Optimum Temperature: Temperature at which its

growth rate is the fastest.

• Maximum Temperature: Temperature above which

growth ceases, or highest temperature at which
microbes will grow.

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Classification of Microorganisms
by Temperature Requirements

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 Temperature Classes of Organisms
• Psychrophiles ( 0-20OC)
– Cold temperature optima
– Most extreme representatives inhabit permanently cold
environments
• Mesophiles ( 20 – 45oC)
– Midrange temperature optima
– Found in warm-blooded animals and in terrestrial and aquatic
environments in temperate and tropical latitudes

• Thermophiles ( 50- 80OC)

– Growth temperature optima between 45ºC and 80ºC

• Hyperthermophiles
– Optima greater than 80°C
– These organisms inhabit hot environments including boiling hot
springs, as well as undersea hydrothermal vents that can have
temperatures in excess of 100ºC

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 pH and Microbial Growth

pH – measure of [H+]
each organism has a pH range and a pH optimum
acidophiles – optimum in pH range 1-4
alkalophiles – optimum in pH range 8.5-11
• lactic acid bacteria – 4-7
• Thiobacillus thiooxidans – 2.2-2.8
• fungi – 4-6

internal pH regulated by BUFFERS and near neutral

adjusted with ion pumps
Human blood and tissues has pH 7.2+0.2

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 pH and Microbial Growth
• The acidity or alkalinity of an environment can greatly
affect microbial growth.
• Most organisms grow best between pH 6 and 8, but
some organisms have evolved to grow best at low or high
pH. The internal pH of a cell must stay relatively close to
neutral even though the external pH is highly acidic or
basic.
– Acidophiles : organisms that grow best at low pH
(Helicobacter pylori, Thiobacillus thiooxidans )
– Alkalophiles : organisms that grow best at high pH
(Vibrio cholera)
– Most of pathogenic bacteria are neutrophiles

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 Osmotic pressure
Osmotic Effects on Microbial Growth
• Osmotic pressure depends on the surrounding solute
concentration and water availability
• Water availability is generally expressed in physical terms
such as water activity (aw)
• Water activity is the ratio of the vapor pressure of the air
in equilibrium with a substance or solution to the vapor
pressure of pure water ( aw 1.00).
aw= P solute
P water

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Osmotic Effect and water activity

organisms which thrive in high solute – osmophiles

organisms which tolerate high solute – osmotolerant
organisms which thrive in high salt – halophiles
organisms which tolerate high salt – halotolerant
organisms which thrive in high pressure – barophiles
organisms which tolerate high pressure – barotolerant

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 Osmotic Pressure Effect
• Hypertonic environments, or an increase in
salt or sugar, cause plasmolysis
• Extreme or obligate halophiles require high
osmotic pressure
• Facultative halophiles tolerate high osmotic
pressure

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Plasmolysis

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Halophiles and Related Organisms
• In nature, osmotic effects are of interest mainly in
habitats with high salt environments that have
reduced water availability

• Halophiles : have evolved to grow best at reduced

water potential, and some (extreme halophiles e.g.
Halobacterium, Dunaliella ) even require high levels
of salts for growth.

• Halotolerant : can tolerate some reduction in the

water activity of their environment but generally
grow best in the absence of the added solute

• Xerophiles : are able to grow in very dry

environments 13
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 Microbial Nutrition
• Why is nutrition important?

– The hundreds of chemical compounds

present inside a living cell are formed from
nutrients.

• Macronutrients : elements required in fairly

large amounts
• Micronutrients : metals and organic
compounds needed in very small amounts
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 Macronutrients

• 95% or more of cell dry weight is made up of a few

major elements: carbon, oxygen, hydrogen,
nitrogen, sulfur, phosphorus, potassium, calcium,
magnesium and iron.
• The first six ( C, H, O, N, P and S) are components
of carbohydrates, lipids, proteins and nucleic acids

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 Main Macronutrients

• Carbon (C, 50% of dry weight) and nitrogen (N, 12% of

dry weight)
• Autotrophs are able to build all of their cellular organic
molecules from carbon dioxide
• Nitrogen mainly incorporated in proteins, nucleic acids
• Most Bacteria can use Ammonia -NH3 and many can
also use NO3-
• Nitrogen fixers can utilize atmospheric nitrogen (N2)

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 Microbial growth requirements

• Source of carbon for basic structures

• Source of cellular energy (ATP or related
compounds) to drive metabolic reactions
• Source of high energy electrons/H, reducing
power, typically in form of NADH/NADPH

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Classification of organisms based on sources
of C and energy used

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 Photoautotroph
Algae, Cyanobacteria
Light + Chlorophyll
CO2 + H2O （CH2O） +O2

Purple and green bacteria

Light + bacteriochlorophyll
CO2 + 2H2S （CH2O） + H2O + 2S

Photoheterotroph
Purple nonsulfur bacteria (Rhodospirillum)

CO2 + 2CH3CHOHCH3 Light + bacteriochlorophyll （CH2O) + H2O + 2CH3COCH3

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 Nitrogen requirements
• Although many biological components
within living organisms contain N, and N2 is
the most abundant component of air, very
few organisms can “fix” or utilize N2 by
converting it to NH3
• N is often growth limiting as organisms
must find source as NH4+ for biosynthesis
• Photosynthetic organisms and many
microbes can reduce NO3- to NH4+

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 Other Macronutrients
• Phosphate (P), sulfur (S), potassium (K), magnesium
(Mg), calcium (Ca), sodium (Na), iron (Fe)
• Iron plays a major role in cellular respiration, being a
key component of cytochromes and iron-sulfur
proteins involved in electron transport.
• Siderophores : Iron-binding agents that cells produce
to obtain iron from various insoluble minerals.

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 Trace Elements

Microbes require very small amounts of other

mineral elements, such as iron, copper,
molybdenum, and zinc; these are referred to as
trace elements. Most are essential for activity of
certain enzymes, usually as cofactors.

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 Growth Factors
(1)Amino acids

(2) Purines and pyrimidines,

(3) Vitamins

Amino acids for protein synthesis

Purines and pyrimidines for nucleic acid synthesis.
Vitamins are small organic molecules that usually make
up all or part enzyme cofactors, and only very small
amounts are required for growth.
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 Properties of microbial photosynthetic systems

Property Cyanobacteria Green and purple Purple nonsulfur

bacteria bacteria

Photo - Chlorophyll Bacteriochlorophyll Bacteriochlorophyll

pigment
O2 production Yes No No
Electron donors H2O H2, H2S, S H2, H2S, S
Carbon source CO2 CO2 Organic / CO2
Primary
products of ATP + NADPH ATP ATP
energy
conversion

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 Chemoautotroph
Bacteria Electron Electron Products
donor acceptor
Alcaligens and H2 O2 H2O
Pseudomonas sp.
Nitrobacter NO2- O2 NO3- , H2O
Nitrosomonas NH4+ O2 NO2- , H2O
Desulfovibrio H2 SO4 2- H2O. H2S
Thiobacillus denitrificans S0. H2S NO3- SO4 2- , N2
Thiobacillus ferrooxidans Fe2+ O2 Fe3+ , H2O

Nitrifying bacteria

2 NH4+ + 3 O2 2 NO2- + 2 H2O + 4 H+ + 132 Kcal

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 Factors that Influence Growth
• Oxygen concentration
– Strict aerobes: Require oxygen for growth (~20%)
– Strict anaerobes: Grow in the absence of oxygen;
cannot grow in the presence of oxygen
– Facultative anaerobes : Grow best in the presence of
oxygen, but are able to grow (at reduced rates) in the
absence of oxygen
– Aerotolerant anaerobes : Can grow equally well in the
presence or absence of oxygen
– Microaerophiles: Require reduced concentrations of
oxygen (~2 – 10%) for growth

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 Toxic Oxygen
• Singlet oxygen: O2 boosted to a higher-energy
state
• Superoxide free radicals: O2–
O2   O2   2H  superoxide dismutase
H2O2  O2
• Peroxide anion: O22–
2H2O2 catalase 2H2O  O2
H2O2  2O  peroxidase 2H2O
• Hydroxyl radical (OH•)

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Classification of organisms based on O2 utilization

• Utilization of O2 during metabolism yields toxic

by-products including O2-, singlet oxygen (1O2)
and/or H2O2.
• Toxic O2 products can be converted to harmless
substances if the organism has catalase (or
peroxidase) and superoxide dismutase (SOD)
• SOD converts O2- into H2O2 and O2
• Catalase breaks down H2O2 into H2O and O2
• Any organism that can live in or requires O2 has
SOD and catalase (peroxidase)

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30
 Uptake of nutrients
Nutrient molecules frequently cannot cross selectively permeable
plasma membranes through passive diffusion and must be
transported by one of three major mechanisms involving the use
of membrane carrier proteins.

Mechanism of nutrient transport :

1. Passive transport : simple diffusion
2. Facilitated diffusion
3. Active transport
4. Group translocation

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 1. Passive diffusion

Passive diffusion
 is the process in which molecules move from a
region of higher concentration to one of lower
concentration as a result of random thermal
agitation. Ex: glycerol,

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 2. Facilitated diffusion

 The rate of diffusion across selectively permeable membranes is

greatly increased by the use of carrier proteins, sometimes called
permeases, which are embedded in the plasma membrane.
 Since the diffusion process is aided by a carrier, it is called facilitated
diffusion. The rate of facilitated diffusion increases with the
concentratioti gradient much more rapidly and at lower
concentrations of the diffusing molecule than that of passive
diffusion 33
A model of facilitated diffusion

The membrane carrier can change

conformation after binding an
external molecule and
subsequently release the molecule
on the cell interior. It then returns
to the outward oriented position
and is ready to bind another solute
molecule.

Because there is no energy input, molecules

will continue to enter only as long as their
concentration is greater on the outside.

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 3. Active transport

Active transport is the transport of solute

molecules to higher concentrations, or against a
concentration gradient, with the use of metabolic
energy input. 35
 4. Group translocation

The best-known group translocation system is the

phosphoenolpyruvate: sugar phosphotransferase
system (PTS), which transports a variety of sugars
into procaryotic cells while Simultaneously
phosphorylating them using phosphoenolpyruvate
(PEP) as the phosphate donor.

PEP + sugar (outside) pyruvate + sugar-P (inside)

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 The phosphoenolpyruvate: sugar phosphotransferase
system of E. coli. The following components are
involved in the system: phosphoenolpyruvate, PEP;
enzyme 1, E I; the low molecular weight heat-stable
protein, HPr; enzyme 11, E II,- and enzyme III, E III.

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 Simple comparison of transport systems

Items Passive Facilitated Active Group

diffusion diffusion transport translocation

carrier Non Yes Yes Yes

proteins
transport Slow Rapid Rapid Rapid
speed

against Non Non Yes Yes

gradient

Specificity Specificity Specificity

transport No specificity
molecules
metabolic No need Need Need Need
energy

Solutes Not changed Changed Changed Changed

molecules
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