E R V TH

A P T GROW
CH LI SM A ND
ABO
ME T
IA L
T E R
BAC

9/21
11/1
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 Learning objective:
At the end of this chapter the students will be able to:
0 Explain bacterial nutritional and environmental requirement
0 Discuss Bacterial metabolism
0 Describe bacterial growth and growth curve

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 Chapter V
Bacterial metabolism and growth
Nutrition
0 For optimal growth and multiplication, bacteria requires
nutrients, such as water, energy, carbon, nitrogen and some
inorganic salts.
0 Bacteria also require various environmental factors for growth in
optimum concentration. These include Oxygen/Carbon dioxide,
pH, temperature and light
0 All bacteria need some form of the element Carbon, H, O2, S, P, and
N for growth.
0 Special elements such as K, Ca, Fe, Mn, Mg, Co, Cu, Z, Ur are
needed by certain bacteria.
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 Nutrition … Cont’d

0 Some have specific vitamin, and growth factor requirements

and others need organic substances secreted by other
microorganisms during their growth.
0 In most case, bacteria need small amount of salt concentration
to grow.
0 Halophytes are bacteria which need high concentration of salt for
their growth.

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 1. Nutrient requirement
0 Depending on their nutritional requirement
bacteria can be classified
0 Autotrophs: - are free-living, non-pathogenic
bacteria, most of which can use carbon dioxide as
their carbon source.
0 The energy needed for their metabolism can be
obtained from
0(a). Sunlight-photoautotrophs and
0(b) by oxidation inorganic compounds -
chemoautotrophs

0 Heterotrophs are generally parasitic bacteria which

require more complex organic compounds than
carbon dioxide as their source of carbon and energy,
e.g. sugar
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2. Temperature requirement
0 Most pathogenic bacteria grow best at an optimum
temperature of 370C.
0 Optimum temperature is the temperature at which growth
occur best.
0 Based on temperature requirement, microorganisms can be
broadly classified into
0 Psycrophylic- are those bacteria, which grow in the range of -5
to 200C
0 These bacteria include those which cause spoilages of food at
refrigeration temperature (2-8oc).
0 Mesophilic- are those bacteria, which grow at 20-450C and
show optimum growth at 37oC.
0 all medically important bacteria (pathogenic bacteria) belong to
this group.

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2. Temperature requirement …Cont’d

0 Thermophilic – are those organisms which prefer

high temperature (50-800C)
0 May cause spoilage of under processed canned food
0 Hyperthermophilic
0 Those which grow at a temperature of above 800C
0 Some of them grow even at 2500C
0 are found in hot springs, geysers and industrial heated
wastes

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 Bacterial Growth

Temperature
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 3. Oxygen requirement

The need of oxygen for particular bacterium reflects its

mechanism to meet the requirement of energy. On the basis of
this requirement, bacteria have been divided in to:
0 Obligate Anaerobes-these grow only in the environment devoid
of oxygen
0 e.g. clostridium
0 Facultative aerobes- these can grow under both aerobic and
anaerobic conditions, e.g. enterobacteriaceae
0 Obligate aerobes- these cannot grow unless oxygen is present
in the medium, e.g. pseudomonas
0 Microaerophilic- these organisms can grow under conditions
with low oxygen tension e.g. clostridium titani.
0 Aerotolerant anaerobes – These bacteria oxidize nutrient
substrates without using elemental oxygen. Unlike obligate
anaerobes, they can tolerate the presence of oxygen.
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4. pH requirement

Most pathogenic bacteria require a pH of 7.2-7.6 for their

optimal growth. Based on pH requirement bacteria can be
classified as;

0 Neutrophilic:- bacteria grow best at neutral pH (pH=7)

0 Most pathogenic micro-organism best grow at neutral pH (pH=7)
0 Acidophilic
0 Bacterial grow best at acidic pH (pH<7)
0  E.g. Lactobacilli, fungi and yeast
0 Alkalophilic
0 Bacterial grow best at Alkaline pH (pH>7)
0 E.g. Vibrio cholerae grow at a pH of 8.6

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Bacterial Metabolism

Bacterial metabolism involves all the cellular processes required

for the survival and replication of the organism.
It refers to all of the chemical reactions occurring within a cell,
including the production of energy, intermediate products, and
end products.
A metabolite is any molecules that are a nutrient (energy source),
intermediary product, or end product in a metabolic reaction.
Most biochemical reactions fall into two categories: Catabolism
and Anabolism.

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Catabolism
0 The metabolic degradation (breakdown) of organic compounds
that results in the production of energy and smaller molecules.
0 Catabolic reactions involve the breaking of bonds; whenever
chemical bonds are broken, energy is released
Anabolism
0 Refers to those biosynthetic processes that use energy for the
synthesis of protoplasmic materials needed for growth,
maintenance, and other cellular functions.
0 Anabolic reaction involves the creation of bonds; it takes energy to
create chemical bonds.
0 Smaller molecules are bonded together to create large molecules

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Heterotrophic Metabolism
0 the biologic oxidation of organic compounds, such as glucose, to yield ATP and
simpler organic (or inorganic) compounds, which are needed by the bacterial
cell for biosynthetic or assimilatory reactions.
Respiration
0 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.
(An additional 308,000 cal is lost as heat.)
Fermentation
0 another type of heterotrophic metabolism,
0 an organic compound rather than oxygen is the terminal electron (or hydrogen)
acceptor.
0 Less energy is generated from this incomplete form of glucose oxidation, but the
process supports anaerobic growth.
Autotrophy
0 a unique form of metabolism found only in bacteria.
0 Inorganic compounds are oxidized directly to yield energy (e.g., NH3, NO2-, S2,
and Fe2+).
Bacterial Photosynthesis
0 a light-dependent, anaerobic mode of metabolism.
0 Carbon dioxide is reduced to glucose, which is used for both biosynthesis and
energy
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 Metabolic pathways
Krebs Cycle
 is the oxidative process in respiration by which pyruvate (via acetyl coenzyme
A) is completely decarboxylated to C02.
 The pathway yields 15 moles of ATP (150,000 calories).
Glyocxylate Cycle
 occurs in some bacteria,
 is a modification of the Krebs cycle.
 Acetyl coenzyme A is generated directly from oxidation of fatty acids or other
lipid compounds.

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Cont…
The Nitrogen Cycle
consists of a recycling process by which organic and inorganic
nitrogen compounds are used metabolically and recycled
among bacteria, plants, and animals.

Important processes, including ammonification,

mineralization, nitrification, denitrification, and nitrogen
fixation, are carried out primarily by bacteria.(Reading
assignment)
Electron Transport and Oxidative Phosphorylation
In the final stage of respiration, ATP is formed through a series
of electron transfer reactions within the cytoplasmic
membrane that drive the oxidative phosphorylation of ADP to
ATP.

Bacteria use various flavins, cytochrome, and non-heme iron

components as well as multiple cytochrome oxidases for this
process
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Bacterial growth
0 Bacteria divide by binary fission.
0 When a bacterial cell reaches a certain size, it divides to form
two daughter cells.
0 Nuclear division precedes cell division and, therefore, in a
growing population, many cell carrying two nuclear bodies can
be found.

Generation time or population doubling time.

0 The interval of time between two cell division, or the time
required for a bacterium to give rise to two daughter cells
under optimum conditions

0 The generation time of bacteria ranges from as little as 20

minutes for E-coli to more than 20 hrs for Mycobacterium
tuberculosis.

The generation time varies not only with the species but also
with the amount of nutrients, the temperature, the pH, and
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other environmental factors.

 Bacterial growth curve
The growth cycle of bacteria has four major phases.
If a small number of bacteria are inoculated into a liquid
nutrient medium and the bacteria are counted at frequent
interval, the typical phase of a standard growth curve can be
demonstrated.
1. The Lag Phase
this phase is of short duration in which bacteria adapt
themselves to new environment in such away that the
bacterial machinery brings itself in conformity with the
nutrition available.
This is a period of active macro molecular synthesis like DNA,
RNA, various enzymes and other structural components
It is the preparation time for reproduction
No increase in cell number occurs, however, vigorous
metabolic activity occurs.
This can last for a few minutes up to many hours.
The duration of lag phases varies with the species, nature of
culture medium, temperature of incubation etc.
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 Bacterial growth curve …Cont’d
2. The log, logarithmic, or exponential phase
0 During this phase, the population can double
approximately every 30 minutes with fast growing
bacteria

0 It has limited duration because of:-

0 Exhaustion of nutrients
0 Accumulation of toxic metabolic end products
0 Rise in cell density
0 Change in pH and
0 Decrease in oxygen tension (in case of aerobic organisms)

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Bacterial growth curve …Cont’d

0 The number of bacteria during log phase growth can

be calculated by the following equation
0 Nt= No x 2 t/d
0 Nt = is the number of bacteria after time (t),
0 t/d = is the amount of time divided by the
doubling time
0 No = the initial number of bacteria;

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Bacterial growth curve …Cont’d
3. Stationary Phase
0 Occur when nutrients depletion or toxic products cause growth to
slow until the number of new cells produced balances the number
of cells that die resulting in a steady state
0 The number of viable cell remain constant
0 There is almost a balance between the bacterial reproduction and
bacterial death
The death/decline phase
0 Due to depletion of nutrients and accumulation of toxic end
products the number of bacteria dying is much more than those
dividing and hence there is gradual decline in the total number of
organism.
0 There is drastic decline in viable cells.

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11/19/21 Fig. Bacterial growth curve 21
 Bacterial Growth curve
0 Morphological & Physiological alterations during growth
 Lag phase – maximum cell size towards the end of lag
phase.
 Log phase – smaller cells, stain uniformly
 Stationary phase – irregular staining, sporulation and
production of exotoxins & antibiotics
 Phase of Decline –involution forms (with ageing)

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 Bacterial Growth curve

Maintenance of cells in the exponential phase

 Cells can be maintained in Log phase by transferring them
repeatedly into fresh continuous culture of identical composition.
 Cultures that require a continuous supply of the cell suspension or
the product in the medium are known as continuous cultures.

 This process can be carried out in 2 devices.

1. Chemostat
 Consists of a culture vessel, overflow siphon & a mechanism for
dripping in fresh medium from a reservoir at a regulated rate.
 Cell concentration remains constant.

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 Bacterial Growth
2. Turbidostat
 The addition of fresh medium is triggered when the turbidity
in the growth vessel reaches a predetermined value that is
monitored by a photoelectric cell.
 As a result, the cells are diluted and continue to grow until
they again reach the critical turbidity

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11/19/21 Fig. continuous-culture device 25