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
 Growth slower as nutrients
decline
 Easy to set up and maintain
 Only one batch lost if
contamination occurs
 Less efficient, fermenter not
always in use
 Useful for producing
secondary metabolites
Continuous
 Growth faster as always
nutrients there
 Difficult to set up and
maintain
 Huge volumes lost if
contamination occurs
 More efficient fermenter in
use constantly
 Useful for producing primary
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.