MICROBIAL:

GROWTH KINETICS &

METABOLITES
-In Batch System-
 GROWTH KINETICS
 deals with the rate of cell growth
 important for the design and operation of
fermentation systems employing them

 An ideal culture for fermentation should (be):

1. pure.
2. grow and reproduce quickly.
3. genetically stable yet easy to manipulation for
better performance.
4. produce uniform product in a short time.
5. not produce undesirable by-products.
6. have a protective mechanism against other
undesirable contaminants.
Growth Curve
 Generating time  time required for cell to divide and reach
double in population
 Generating time equivalent to the average length of the cell
cycle
 System culture: batch & continues
 BATCH:
 Closed single culture vessel  without any inlet and outlet

 No fresh medium is provided during incubation

 CONTINUES:
 Open system

 Fresh media continuestly added to the culture  constant

rate & old medium is removed at the same rate

BATCH SYSTEM
 close system, without any inlet or outlet
streams
 nutrients are fixed amount limited
 The inocula are transferred, then
gradually grow and replicate
 As the cell propagates, the nutrients are
depleted and end products are formed
 Main stages of a growth curve: lag,
exponential, stationary & death phases
Cell growth curve
 this description refers to
the behaviour of both
unicellular and mycelial
(filamentous) organisms in
batch culture
 The growth of mycelial
resulting in the exponential
addition of viable biomass
to the mycelial body rather
than the production of
separate, discrete unicells.
Lag phase
 the cell number does not increase
 the cells may grow in size
 duration of time for adaptation of
microorganisms to the new environment,
without much cell replication and with no sign
of growth.
 shock to the environment when there is no
acclimation period
 Length of lag phase  inoculum (concentration, type,
age), medium composition, fermentation conditions

i) size of the inoculum

If a small amount of cells are inoculated into a large
volume  a long lag phase.
ii) medium
 Transfer microorganisms from a low nutrient to high
concentration long lag period, because the cells
must produce the enzymes necessary for
themetabolization of the available nutrients.
 If they are moved from ahigh to a low nutrient
concentration short lag phase
Short lag phase
 Excessive lag phase unproductive
 Minimize lag phase period:

1. the composition of the medium and the

environmental conditions in the seed culture and
the production vessel are identical
2. the dilution shock is small (i.e. a large amount of
inoculum is used)
3. the cells in the inoculum are in the late
exponential phase of growth.
Exponential phase
The stages:
i) Accelerated growth phase: The cell number starts to
increase and the division rate increases to reach a
maximum sometimes included as part of lag phase
ii) Exponential growth phase: The cell number increases
exponentially as the cells start to divide
 Plotting the linear increase growth in semi-log graph
shown a constant slope
 Slope representing a constant rate of cell population

ii) Decelerated growth phase: After the growth rate

reaches a maximum, it is followed by the deceleration
of both growth rate and the division rate
 Primary metabolism products in tropophase periode
Stationary phase
 The cell population will reach a maximum value
 will not increase any further  growth rate zero
 cell density remains constant
 The growth of microbial populations is normally limited
either by the exhaustion of available nutrients or by
the accumulation of toxic products of metabolism 
the rate of growth declines and growth eventually
stops
 The transition between the exponential phase and the
stationary phase involves a period of unbalanced
growth during which the various cellular components
are synthesized at unequal rates.
 Consequently, cells in the stationary phase have a
chemical composition different from that of cells in the
exponential phase
 However, in this phase metabolisms are
still active
 Produce compounds are not synthesized
during tropophase (exp. Phase)
secondary metabolism, no obvious
function in cell metabolism
idiophase
 employ primary products as raw material
 very few microorganism species; not all
Death phase
 activities of the cell gradually decrease as they age
 In the end of stationary phase  cell may start to die
 system is dead
 even if you add food, you will get no growth.
 Death occurs either because of the depletion of the
cellular reserves of energy, or the accumulation of toxic
products  deactivating remaining cells
 the cell growth rate balances the death rate.
 In some cases, the organisms not only die but also
disintegrate, a process called lysis.
 a death phase develops while the cell density drastically
drops if the toxic secondary metabolites are present
 exponential decrease in the number of living cells in the
media while nutrients are depleted.
CONTINUOUS SYSTEM
 microorganisms grow in a system with constant
environmental conditions maintained through continual
provision of nutrients and removal of wastes
 Typically the concentration of cells will reach an
equilibrium level that remains constant as long as the
nutrient feed is maintain
 maintain a microbial population in exponential growth
 growing at a known rate and at a constant biomass
concentration for extended periods.
 Continuous culture systems make possible the study of
microbial growth at very low nutrient levels,
concentrations close to those present in natural
environments.
 Two major types of continuous culture systems
commonly are used: chemostats and turbidostats.
 Chemostat
 The culture medium for a chemostat possesses an
essential nutrient (e.g., a vitamin) in limiting
quantities.
 Because one nutrient is limiting, growth rate is
determined by the rate at which new medium is fed
into the growth chamber;
 the final cell density depends on the concentration
of the limiting nutrient
 Both population size is related to the dilution rate
 population density remains unchanged over a wide
range of dilution rates
 The generation time decreases (i.e., the rate of growth
increases) as the dilution rate increases.
 Dilution rate the flow of medium per time (F) over the
volume (V) of culture in the bioreactor
 If the dilution rate rises too high, microorganisms can
actually be washed out of the culture vessel before
reproducing because the dilution rate is greater than the
maximum growth rate.
 This occurs because fewer microorganisms are present to
consume the limiting nutrient.
 When dilution rates are low, only a limited supply of
nutrient is available and the microbes can conserve only a
limited amount of energy
 As the dilution rate increases, the amount of nutrients and
the resulting cell density rise because energy is available
for both maintenance and reproduction.
 The growth rate increases when the total available energy
exceeds the maintenance energy
Turbidiostat

 has a photocell that

measures the turbidity
(absorbance) of the
culture in the growth
vessel.
 The flow rate of media
through the vessel is
automatically
regulated to maintain
a predetermined
turbidity.
 Because turbidity is
related to cell density,
the turbidostat
maintains a desired
cell density.
The Difference
Turbidostat
 Microbe concentration is kept
a t a preset level by diluting
the culture with fresh
medium
 The dilution rate varies
rather than remaining
constant
 The medium contains all
nutrients in excess  none of
the nutrients is limiting
 operates best at high dilution
rates
 Measures the absorbance or
turbidity of the culture in the
growth vessel
 Automatically regulated to
maintain a predetermined
turbidity or cell density
CHEMOStat
 A flow of fresh medium is
introduced into the culture at
a steady, predetermined rate
 The latter adds a limiting
vital nutrient
 In this way the growth rate
and not the cell density is
kept constant
 most stable and effective at
lower dilution rates
 Rate of growth can be
controlled either by
controlling the rate at which
new medium enters the
growth chamber or by
limiting a required growth
factor in the medium
 Compare and contrast the processes of
continuous and batch culture.

Batch Continuous
 Growth slower as
 Growth faster as always
nutrients decline nutrients there
 Difficult to set up and
 Easy to set up and
maintain
maintain
 Huge volumes lost if
 Only one batch lost if contamination occurs
contamination occurs
 More efficient fermenter
 Less efficient, fermenter in use constantly
not always in use  Useful for producing
 Useful for producing primary metabolites
secondary metabolites
© Pearson Education Ltd 2009
This document may have been altered from the original
MICROBIAL METABOLITES
 Metabolism the sum of all the biochemical
reactions carried out by an organism.
 The kinetic description of batch culture may be
rather misleading when considering the product-
forming capacity of the culture during the various
phases
 Bu’Lock proposed a descriptive terminology of
the behaviour of microbial cells which considered
the type of metabolism rather than the kinetics of
growth
 Tropophase & iodophase
 Tropophase
 describes the log or exponential phase of a culture
 which the sole products of metabolism are either
essential to growth, development, and reproduction 
essential for survive (such as amino acids,
nucleotides, proteins, nucleic acids, lipids,
carbohydrates)
 It usually performs a physiological function in the
organism (i.e. an intrinsic function)
 Or are the by-products of energy-yielding metabolism
such as ethanol, acetone and butanol.
 The metabolites produced during the trophophase are
referred to as primary metabolites.

 products which do not have
an obvious role in cell
metabolism.
 The metabolites produced
during the idiophase are
referred to as the secondary
metabolites.
 Secondary metabolites are
produced when the cell is
not operating under
optimum conditions., when
primary nutrient source is
depleted.
 Secondary metabolites tend
to be synthesized from the
intermediates and end-
products of primary
metabolism.
Idiophase
 Secondary metabolites are
synthesized for a finite
period by cells that are no
longer undergoing balanced
growth
 Although the primary
metabolic routes are
common to the vast majority
of microorganisms, each
secondary metabolite would
be synthesized by very few
microbial taxa
 not all microbial taxa
undergo secondary
metabolism;
 it is a common feature of the
filamentous fungi and
bacteria
 Although taxonomic distribution
of secondary metabolism is far
more limited than that of
primary metabolism, the range
of secondary products produced
is enormous
 It is sometimes difficult to
categorize a product as primary
or secondary, and the kinetics
of production of certain
compounds may change,
depending on the growth
conditions employed
Relation between 1 & 2
 Primary metabolic pathway  the synthesis of aromatic amino
acids
 The secondary metabolite  antibiotics containing aromatic rings.
The inter-relationships between
primary and secondary metabolism
 3 major product of secondary metabolism categories:
alkaloids, essential oil and glycosides
1. Alkaloids:
N-containinng compound  used as phaarmaceutical
industries
 E.g: codein, nicotine, caffeine, morphine

2. Essential oil
mixtures of terpenoid  used as flavorents, fragrance
and solvents
3. Glycosides
includes phenolics, tannins & flavonoid, saponins, &
cyanogenic glycosides  used as dye, flavors,
pharmaceuticals etc.