CHAPTER V

BACTERIAL METABOLISM AND

GROWTH
 Learning objective:
At the end of this chapter the students will be able to:
• Explain bacterial nutritional and environmental requirement
• Discuss Bacterial metabolism
• Describe bacterial growth and growth curve
 Chapter V
Bacterial metabolism and growth
Nutrition
• For optimal growth and multiplication, bacteria requires
nutrients, such as water, energy, carbon, nitrogen and some
inorganic salts.
• Bacteria also require various environmental factors for growth
in optimum concentration. These include Oxygen/Carbon
dioxide, pH, temperature and light
• All bacteria need some form of the element Carbon, H, O2, S,
P, and N for growth.
• Special elements such as K, Ca, Fe, Mn, Mg, Co, Cu, Z, Ur
are needed by certain bacteria.
 Nutrition … Cont’d

• Some have specific vitamin, and growth factor requirements

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

• Depending on their nutritional requirement bacteria can be

classified
– Autotrophs: - are free-living, non-pathogenic bacteria,
most of which can use carbon dioxide as their carbon
source.
• The energy needed for their metabolism can be
obtained from
– (a). Sunlight-photoautotrophs and
– (b) inorganic compounds by oxidation-
chemoautotrophs
– Heterotrophs are generally parasitic bacteria which
require more complex organic compounds than carbon
dioxide as their source of carbon and energy, e.g. sugar
 2. Temperature requirement

• Most pathogenic bacteria grow best at an optimum

temperature of 370C.
• Optimum temperature is the temperature at which growth
occur best.
• Based on temperature requirement, microorganisms can be
broadly classified into
• Psycrophylic- are those bacteria, which grow in the range
of -5 to 200C
– These bacteria include those which cause spoilages of
food at refrigeration temperature (2-8oc).
• Mesophilic- are those bacteria, which grow at 20-450C and
show optimum growth at 37oC.
– all medically important bacteria (pathogenic bacteria)
belong to this group.
 2. Temperature requirement …Cont’d

• Thermophilic – are those organisms which prefer high

temperature (50-800C)
– May cause spoilage of under processed canned food
• Hyperthermophilic
– Those which grow at a temperature of above 800C
– Some of them grow even at 2500C
– are found in hot springs, geysers and industrial
heated wastes
 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:
• Obligate Anaerobes-these grow only in the environment
devoid of oxygen
– e.g. clostridium
• Facultative aerobes- these can grow under both aerobic and
anaerobic conditions, e.g. enterobacteriaceae
• Obligate aerobes- these cannot grow unless oxygen is present
in the medium, e.g. pseudomonas
• Microaerophilic- these organisms can grow under conditions
with low oxygen tension e.g. clostridium titani.
• Aerotolerant anaerobes – These bacteria oxidize nutrient
substrates without using elemental oxygen. Unlike obligate
anaerobes, they can tolerate the presence of oxygen.
 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
• Neutrophilic:- bacteria grow best at neutral pH (pH=7)
– Most pathogenic micro-organism best grow at neutral pH
(pH=7)
• Acidophilic
– Bacterial grow best at acidic pH (pH<7)
–  E.g. Lactobacilli, fungi and yeast
• Alkalophilic
– Bacterial grow best at Alkaline pH (pH>7)
– E.g. Vibrio cholerae grow at a pH of 8.6
 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.
Catabolism
• The metabolic degradation (breakdown) of organic
compounds that results in the production of energy and
smaller molecules.
• Catabolic reactions involve the breaking of bonds; whenever
chemical bonds are broken, energy is released
Anabolism
• refers to those biosynthetic processes that use energy for the
synthesis of protoplasmic materials needed for growth,
maintenance, and other cellular functions.
– Anabolic reaction involves the creation of bonds; it takes
energy to create chemical bonds.
– Smaller molecules are bonded together to create large
molecules
Heterotrophic Metabolism
• 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.
Autotrophy
• a unique form of metabolism found only in bacteria.
• Inorganic compounds are oxidized directly (without using sunlight) to yield
energy (e.g., NH3, NO2-, S2, and Fe2+).
Respiration
• 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
• 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.
Bacterial Photosynthesis
• a light-dependent, anaerobic mode of metabolism.
• Carbon dioxide is reduced to glucose, which is used for both biosynthesis
and energy production.
 Metabolic pathways
Krebs Cycle
• is the oxidative process in respiration by which pyruvate (via acetyl coenzyme A) is
completely decarboxylated to C0 2.
• The pathway yields 15 moles of ATP (150,000 calories).
Glyoxylate 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.
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.
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.
 Bacterial growth

• Bacteria divide by binary fission.

• When a bacterial cell reaches a certain size, it divides to
form two daughter cells.
• 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.
• 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
• 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
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.
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.
 Bacterial growth curve …Cont’d

The log, logarithmic, or exponential phase

• During this phase, the population can double
approximately every 30 minutes with fast growing
bacteria
• It has limited duration because of:-
– Exhaustion of nutrients
– Accumulation of toxic metabolic end products
– Rise in cell density
– Change in pH and
– Decrease in oxygen tension (in case of aerobic
organisms)
 Bacterial growth curve …Cont’d

• The number of bacteria during log phase

growth can be calculated by the following
equation
• Nt= No x 2 t/d
• Nt = is the number of bacteria after time (t),
• t/d = is the amount of time divided by the
doubling time
• No = the initial number of bacteria;
 Bacterial growth curve …Cont’d

Stationary Phase
• 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
• The number of viable cell remain constant
• There is almost a balance between the bacterial reproduction
and bacterial death
The death/decline phase
• 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.
• There is drastic decline in viable cells.
Fig. Bacterial growth curve
Continuous-culture

• to maintain a culture in exponential, steady-state (balanced)

growth for long periods is to use a device in which fresh
medium is continuously added but the total volume of
culture is held constant by an overflow tube.
• One such constant-volume device is called a chemosta
– it operates by infusing fresh medium containing a limiting nutrient at
a constant rate, and the growth rate of the cells is set by the flow rate
• A similar constant-volume device is the turbidostat
– it operates by the infusion of fresh medium by a pump controlled
indirectly by the turbidity of the culture
• Most of the places in which bacteria live on and within our
bodies, in health and disease, provide conditions more
closely resembling those of nutrient-limited continuous
culture devices than of enclosed flasks.
Fig. continuous-culture device