ChE 233 Bioreactor Design

LECTURE 2.7: HOW CELLS GROW IN BATCH BIOREACTOR

Analiza P. Rollon, Ph.D.
Professor
Department of Chemical Engineering
College of Engineering, University of the Philippines

Objectives:

At the end of this lecture and supplemental readings, the students will be able to
 Discuss various phases of cell growth in batch culture and the model that describe them.

Lecture Outline

1. Cellular Metabolism as Redistribution of Energy

1.1.Substrate Utilization, Cell Growth and Product Formation
1.2.True Growth and Overall Growth

2. Batch Reactors
2.1.Batch Reactor Applications
2.2.Cell Growth in Batch Culture: Growth Curve

3. Biomass Yield
4. Growth as Function of Substrate Concentration: The Monod Equation
5. Maintenance Energy
6. Product Formation
7. Oxygen Demand
8. Oxygen Transfer
9. Effect of Temperature
10. Material Balance in Batch Reactor

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1. Cellular Metabolism as Redistribution of Energy

“Simple” organic substances (feedstock that to be converted to desired product or contaminant

substances in waste or wastewater to be treated) fed to a bioprocess reactor is substrate (“food”)
for the microorganism (or microorganisms) inside the bioreactor.
Large molecules (e.g. proteins, lipids, carbohydrates) are first broken down to smaller molecules
via the action of extracellular enzymes. This breaking down (i.e., hydrolysis) of macro-molecules
is carried out by extracellular enzymes. This process may also be through use of strong chemical
catalysts that aid hydrolysis. With enzymes as catalyst, this hydrolysis may occur in the same
reactor where the products of hydrolysis are used by microorganisms as “food” (i.e.,substrate) or
it may be allowed to take place in a separate reactor preceding the one where the products of
hydrolysis (i.e., smaller molecules) can be taken up and degraded by microorganisms.
Inside the microbial cells, the smaller molecules undergo biochemical processes. These processes
comprise cellular metabolism, which is a redistribution of the chemical energy present in the
substrate to energy present in the product formed, energy contained in the new cells (cellular
biomass) and energy used for cellular maintenance and growth (figure. 1).

Production Product (“waste”)

Cell functioning
CO2, H2O, heat
& maintenance
Substrate

Growth New cells (biomass)

Figure 1. Cellular metabolism as a redistribution of energy

Cells need energy for:

• polymerization (growth)
• biosynthesis (growth)
• formation of precursor metabolites (growth)
• bringing nutrients, substrates into the cell (maintenance)
• keeping metabolites in the cell (maintenance)
• maintaining proper turgor pressure (maintenance)
• maintaining internal pH (maintenance)
• motility (maintenance)

The energy used for above (growth and maintenance) is derived from the energy provided by ATP.
When ATP  ADP + Pi (phosphate): energy is released and allows work-requiring process to
happen.

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ATP used for the above growth and maintenance requirement are ATP formed in substrate-level
phosphorylation or electron transport chain.
In ethanol fermentation, the fungus grows while it coverts sugar in the feed to ethanol. In biological
wastewater treatment, microorganisms grow as they consume organic contaminants in the
wastewater and convert them to oxidized form and product, which is still of higher energy level.
As in any chemical reaction, reactants react and products are formed at definite proportions. In a
balanced chemical reaction, these proportions are indicated by the respective stoichiometric
coefficients of the reactants and products. Examples of balanced biochemical reactions are shown
in previous lectures. These balanced biochemical reactions show the amount of cells
(microorganisms or biomass) formed and the amount of products produced per unit amount of
substrate consumed. These proportions are known as yield of biomass on substrate (YSX) and yield
of product on substrate (YSP).
YSX ≡ amount of biomass produced per unit amount of substrate consumed.
YSP ≡ amount of product produced per unit amount of substrate consumed.
The relations for several types of bioprocesses are discussed below.

1.1.Substrate Utilization, Cellular Growth and Product Formation

Aerobic growth on a single carbon source and energy source, a C-free nitrogen source,
no formation of products.
1
 rS  rX  mS X Eqn. 1.1
YSX
where
rS ≡ substrate utilization rate (mol substrate.m-3.d-1)
rX ≡ biomass production rate (mol biomass.m-3.d-1)
mS ≡ maintenance coefficient (mol substrate.mol-1 biomass.d-1)
X ≡ biomass concentration (mol biomass.m-3)
The rate of biomass production (rX) is a function of the specific growth rate () and the biomass
concentration (X).
rX  X Eqn. 1.2
where
 ≡ specific growth rate, i.e., amount of biomass formed per unit amount of biomass
present (d-1)
When there is product formation:
1 1
 rS  rX  rP  mS X Eqn. 1.3
YSX YSP

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where
rP ≡ product formation rate (mol product.m-3.d-1)
The rate of product formation (rX) can be expressed as function of the specific product formation
rate (qP) and the biomass concentration (X).
 rP  q P X Eqn. 1.4
where
qP ≡ specific product formation rate, i.e., amount of product formed per unit amount of
biomass present (mol product.mol-1 biomass.d-1)

Yield parameters are relatively independent of pH, medium composition and growth rate of the
cells. Thus, yield values determined under certain conditions of pH, etc., can be applied to other
conditions. For example, yield values for suspended cells can be taken as equal to the yield for
immobilized cells, as in biofilms. Theoretical yield values are derived from biochemical
pathways and stoichiometry.

1.2.True Yield and Overall Yield on Substrate

Overall yield
The overall yield for biomass when there is no product formation:
 rX X
YSXOV   Eqn. 1.5
rS 
X  mS X
YSX

Multiplying by YSX/:
YSX
YSXOV  Eqn. 1.6
Y
1  m S SX


The influence of several parameters can be illustrated by performing simulations with a set of
data. For example:
 = 0.01 h-1
YSX = 0.55 kg.kg-1
mS = 0.025 kg.kg-1.h-1

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When the growth becomes too small ( << mS), too much substrate is used for maintenance and
the overall yield starts to decrease. If maintenance requirement can be neglected, the overall
yield YOVSX = YSX.
The maintenance and yield can be varied only in a limited range. The influence of these two
parameters is therefore limited.

The overall yield for product on substrate is:

 rP rP
YSXOV   Eqn. 1.7
rS rX r
 P  mS X
YSX YSP
This overall yield is different from the true yield. Te previous equation can be rewritten in terms
of the specific product formation rate, qP.
 rP YSP
YSXOV   Eqn. 1.8
rS YSP Y
1  mS SP
q P YSX qP

The influence of several parameters can be illustrated by performing simulations with a set of
data. For example:
 = 0.01 h-1
YSX = 0.55 kg.kg-1
YSP = 0.55 kg.kg-1
qP = 0.010 kg.kg-1.h-1
mS = 0.025 kg.kg-1.h-1
Homework 1. Draw a plot of the overall yield at different values of the pertinent parameters.

Note that:
 The specific product formation rate, qP, is the main important variable that influences the
overall yield.
 When qp decreases, the effect of the maintenance and growth increases, reducing the overall
yield of product on substrate.
 When the specific growth rate increases, the overall yield decreases.
 The parameters mS and YSX can only be varied in a limited range. They do not have so much
effect on overall yield for product.

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Specific growth rate
The specific growth of the microorganism is dependent on the substrate concentration (S) as well
as kinetic parameters (kS and m) that are characteristic of the microorganism.
 S 
   max  Eqn. 1.9
 Ks  S 
where
m = maximum specific growth rate (d-1)
kS = Monod half-saturation constant (mol.m-3). This is the value of S at half the
maximum specific growth rate.
S = substrate concentration (mol.m-3)

Specific product formation rate

The specific product formation rate, qP (mol product.mol biomass-1), can be expressed as
follows:
qP  k PG   k PN Eqn. 1.10
where
kPG = growth associated product formation coefficient, i.e., amount of product form per
unit amount biomass (mol product.mol biomass-1)
kPN = non-growth associated product formation coefficient, i.e., amount of product form
per unit amount biomass (mol product.mol biomass-1)

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2. Batch Reactor (BR)

2.1.BR Process and Application

Application: food processing

pharmaceutical bioprocesses
agricultural bioprocesses
environmental processes

BR is well suited to operate under sterile conditions

 Sterile condition is important in biotechnology.
 Maintaining sterile conditions is not feasible in environmental processes.

closed, well-mixed
(i.e. not continuously fed &
no withdrawal of reactor contents)

At start:
 Nutrients are added and mixed in suspension.
 Inoculation: Viable microorganisms are added.
 Feeding: Input of substrate (that is, organic pollutants present in wastewater)
 Biological process proceeds in time. Microorganisms grow. They feed on substrate. In
wastewater treatment, a consortium of microorganisms grows. The product of one group may
be the substrate for another group. Hence, substrate (i.e., organic pollutants) concentration
decreases while cell (biomass) concentration increases. Products are produced. Gaseous
products leave the liquid or aqueous system while dissolved products accumulate in the liquid
or aqueous system. There is no withdrawal of effluent during the batch process.
 When, needed, BR may be provided with pH and temperature control.
 When the desired biological process requires air, the BR is aerated.

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2.2.Batch Growth Curves

Biological conversion phases in a batch reactor (BR), i.e., batch culture:

A. Lag phase
- cells adapt their-enzymatic equipment to new medium

A cell does not produce at all times large quantities of all enzymes it can produce on the basis of
its genetic material. It produces only those that it actually has to use in a given situation.
 assimilatory enzyme - always required and always produced irrespective of the
composition of medium
 inducible enzyme - produced only when needed - to be produced requires inducer,
e.g. a substrate of the enzyme or a compound structurally similar to its substrate

Production of enzymes also requires ATP + building blocks for protein synthesis.
Synthesis of enzymes which are not required is blocked by Depressor which acts on DNA. The
inducer counteracts repressor, hence, permit the concerned DNA to "order" induced synthesis of
that enzyme.

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Lag phase
 After inoculation, microorganisms need to adjust to the new environment, i.e., to
prepare the enzymatic machinery for the specific cellular functions involve in
metabolism (process of growth  anabolism).
Depending on the composition of nutrients, new enzymes are synthesized, the
synthesis of some other enzymes is repressed, and the internal machinery of cells is
adapted to the new environmental conditions.
 Virtually no growth although metabolic activity is high. There can be increase in cell
mass but no increase in cell number density.
 When the inoculum is small and has a low fraction of cells that are viable, there may
be a pseudo-lag phase, which is a result, not of adaptation, but of small inoculums
size or poor condition of the inoculum

Factors affecting length of lag phase:

1. Low concentration of some nutrients and growth factors may also cause a long lag phase.
Examples:
o The lag phase of Enterobacter aerogenes (formerly Aerobacter aerogenes) grown
in glucose and phosphate buffer medium increases as the concentration of Mg2+
(which is an activator of the enzyme phosphatase), is decreased.
o Even heterotrophic cells require CO2 fixation (to supplement intermediates
removed from key energy-producing metabolic cycles during rapid biosynthesis),
and excessive sparging can remove metabolically generated CO2 too rapidly for
cellular restructuring to be accomplished efficiently, particularly with a small
inoculum.

2. The age of the inoculum culture has a strong effect on the length of lag phase.

Age refers to how long a culture has been maintained in a batch culture.
Usually, the lag period increases with the age of the inoculum. In some cases, there is an
optimal inoculums age resulting in minimum lag period.
To minimize the duration of the lag phase, cells should be adapted to the growth medium and
conditions before inoculation, cells should be young (or exponential phase cells) and active,
and the inoculum size should be large (5% to 10% by volume).
The nutrient medium may need to be optimized, and certain growth factors are included to
minimize the lag phase.
Multiple lag phases may be observed when the medium contains more than one carbon source.
This phenomenon, known as diauxic growth, is caused by a shift in metabolic pathways in the
middle of a growth cycle. After one carbon source is exhausted, the cells adapt their metabolic
activities to utilize the second carbon source. The first carbon source is more readily utilizable than
the second, and the presence of more readily available carbon source represses the synthesis of the
enzymes required for the metabolism of the second substrate.

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B. Exponential growth phase (a.k.a. logarithmic growth phase or maximum growth phase)

Exponential or logarithmic growth phase

 Once adjusted to their environment, organisms start to grow. After the adaptation period,
cells can multiply rapidly with maximum rate, and cell mass and cell number density
increase exponentially with time.
 With all growth factor plentifully available, growth occurs in exponential rate.
 Cells divide by binary fission at constant rate.
 This is a period of balanced growth, in which all components of a cell grow at the same
rate (pseudo-steady state). That is, the average composition of a single cell remains
approximately constant during this phase of growth. During balanced growth, the net
specific growth rate determined from either cell number or cell mass would be the same.
The specific growth rate is constant, from which a phenomenological model is proposed for
the exponential growth phase (Malthus model):

rx = dX/dt = X (Eqn. 2.1)

. where X = number of living cells per unit volume
 = specific growth rate

The increase in number of cells (or mass) is proportional to the number of cells.
Integral form: ln X = t + C where C = ln X(t=O) (Eqn. 2.2)
X = e(t + C)

The slope of the straight part (logarithmic phase) of the growth curve (X vs, t) represents .
Relation between  and doubling time (td):
at ti , have X1 ln X1 = t, + C
at t2, have X = 2Xl ln 2X1 = t2 + C
---------------------
ln 2 =  (t2-ti) = td

  = (ln 2)/td = 0.69/td or td = 0.69/ (Eqn. 2.3)

The doubling time is also the time required for a new generation of cells to appear during
exponential growth period.

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C. Deceleration growth phase

 growth decelerate due to exhaustion of one or more essential nutrients  nutrients

become growth-limiting
 formation of toxic by-products leading to decrease in 
 Rapidly changing environment  unbalanced growth.
 During unbalanced growth, cell composition and size will change. In the deceleration
phase, the stresses induced by nutrient depletion or waste accumulation cause a
restructuring of the cell to increase the prospects of cellular survival in a hostile
environment. These observable changes are the result of the molecular mechanisms of
repression and induction.
 Modified Malthus model (Verhulst 1844) includes an apparent biomass inhibition term

(Eqn. 2.4)

k is the carrying capacity coefficient.

(Eqn. 2.5)

 growth and death may occur simultaneously

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D. Stationary phase
 End of deceleration growth phase: Growth becomes equal to rate of death  no net
growth
 Even though the net growth rate is zero during the stationary phase, cells are still
metabolically active and produce secondary metabolites. Primary metabolites are growth-
related products and secondary metabolites are non-growth related. In fact, the
production of certain metabolites is enhanced during the stationary phase (e.g. antibiotics,
some hormones) due to metabolite deregulation.
During the course of the stationary phase, one or more of the following phenomena may take
place:
a. Total cell mass concentration may stay constant, but the number of viable cells may
decrease.
b. Cell lysis may occur, and viable cell mass may drop. A second growth phase may
occur, and cells may grow on lysis products of lysed cells (cryptic growth).
c. Cells may not be growing but may have active metabolism to produce secondary
metabolites. Cellular regulation changes when concentrations of certain metabolites
(carbon, nitrogen, phosphate) are low. Secondary metabolites are produced as a result
of metabolite deregulation.
During the stationary phase, the cell catabolizes cellular reserves for new building blocks and
for energy-producing monomers. This is called endogenous metabolism.

The cell must always expend energy to maintain an energized membrane (i.e. proton-motive
force) and transport of nutrients and for essential metabolic functions such as motility and
repair of damage to cellular structures. This energy expenditure is called maintenance energy.

As such, maintenance energy and endogenous metabolism are not limited to the stationary
phase but become dominant in the stationary phase. The maintenance or endogenous
expenditure is just a small fraction of the total cell needs during maximum growth.

When the primary metabolism diminishes as in the stationary phase, the endogenous
metabolism becomes dominant. Often, dead cells lyse, and intracellular nutrients released into
the medium are used by the living organisms during stationary phase.

The reason for termination of growth may be either exhaustion of an essential nutrient or
accumulation of toxic products. If an inhibitory product is produced and accumulates in the
medium, the growth rate will slow down, depending on inhibitor production, and at a certain
level of inhibitor concentration, growth will stop.

Example: Ethanol production by yeast is an example of a fermentation in which the product is

inhibitory to growth. Dilution of toxified medium, addition of an unmetabolizable chemical
compound complexing with the toxin, or simultaneous removal of the toxin would alleviate
the adverse effects of the toxin and yield further growth.

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E. Phase of logarithmic death

Death phase
 Cell decay rate > growth rate  there is net decay!
 Cell lysis prevails, i.e. breaking open of the cell membrane and the subsequent release
of cell contents into the medium.
 Cell lysis reduces cell number until no living cells are left.

The BR environment can be characterized by a continuously changing environment. Nutrient

concentration, physiological state of the microbial cell and mass of intact cells differ from
one moment to another.

 Similar relation that describes logarithmic growth

 Agent for killing the cell ‘hits’ vital target in the living cell
 The number of effective hits is directly proportional to the number of targets (living cell):

The death rate can be thought of as a first-order reaction. Because S is zero, µG is zero
starting from the stationary phase:
rX = dX/dt = kdX (Eqn. 2.6)
Or lnX = kdt + C  Chick’s law (Eqn. 2.7)

For constant volume:

where XS0 = cell concentration at the beginning of the death phase

During the death phase, cells may or may not lyse, and the reestablishment of the culture may
be possible in the early death phase if cells are transferred into a nutrient-rich medium.
In both the death and stationary phases, there is a distribution of properties among individuals
in a population. With a narrow distribution, cell death will occur nearly simultaneously. With
a broad distribution, a fraction of the population may survive for an extended period. It is this
fraction that would dominate the reestablishment of a culture from inoculum derived from
stationary- or death-phase cultures.
 Thus, using an old inoculum may select for variants of the original strain having altered
metabolic capabilities.

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3. Biomass Yield

 A growth yield accounts for the substrate consumption in the first two terms (right hand
side of above equation) would be more logical.
 Maintenance needs, are required whether the cell is growing or simply staying functional
(or viable), although they can be growth rate dependent in some fashion.
 Conversion to extracellular products may not be directly related to the cell biomass
growth, although it is at least related to maintenance needs.

 For organisms growing aerobically on glucose, YFX/S is typically 0.4 - 0.6 g/g for most
yeast and bacteria, while YFX/O2 is 0.9 - 1.4 g/g.
 Anaerobic growth is less efficient, and the yield factor is reduced substantially.
 With substrates that are more or less reduced than glucose, the value of the apparent yield
will change. For methane, YFX/S would assume values of 0.6 - 1.0 g/g, with the
corresponding YFX/O2 decreasing to about 0.2 g/g. In most cases, the yield of biomass on a
carbon-energy source is 1.0 - 0.4 g biomass per gram of carbon consumed.

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 4. Growth Rate as Function of Substrate Concentration

Growth rate of microorganisms is a function of substrate concentration. An example of the

growth-substrate model is the Monod Equation:

where:
 μ is the specific growth rate of the microorganisms
 μmax is the maximum specific growth rate of the microorganisms
 S is the concentration of the limiting substrate for growth
 Ks is the "half-velocity constant"—the value of S when μ/μmax = 0.5

The Monod equation is a

mathematical model for
the growth of
microorganisms. It is
named for Jacques Monod
who proposed using an
equation of this form to
relate microbial growth
rates in an aqueous
environment to the
concentration of a limiting
nutrient.

At low S such that S <<< Ks: First order (constant: µmax/Ks). At high S such that S >>> Ks: Zero
order (µ = µmax)

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5. Maintenance Energy

Maintenance coefficient – describes the specific rate of substrate uptake for cellular
maintenance, or

During the stationary phase, where little substrate is available, endogenous metabolism of
biomass components is used for maintenance energy.

Cellular maintenance represents energy used to:

 Repair damaged cellular components
 Transfer some nutrients and products in and out of cells,
 Motility
 Adjust osmolarity of the cells interior volume

Note:

 A microbial product formed in association with cell growth is called primary

metabolite. A product formed independent of cell growth is called a secondary
metabolite.
 Polymeric products are referred to by a variety of names: Extracellular
Polymeric Substances (EPS), glycocalix, and extracellular products or
exopolymer.
 The phase of product formation is called idiophase.
 The phase of cell production is called tropophase.
 The idiophase and tropophase may or may not coincide.

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6. Product Formation

Microbial products are classified into three categories:

a. Growth associated product formation

Theses are products produced simultaneously with microbial growth. The specific
rate of product formation is proportional to the specific rate of growth ug.

Examples: constitutive enzymes.

b. Non-growth associated product formation

These are products formed during the stationary phase when the growth rate is zero.
The specific growth rate is constant.

Many secondary metabolites, such as antibiotics (e.g., penicillin), are non-growth

associated products.

c. Mixed growth associated product formation

These are produced during the slow growth and stationary phase.

Examples: Lactic acid, xanthan gum, some secondary metabolites from cell culture.

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7. Oxygen Demand for Aerobic Microorganisms

 Dissolved oxygen (DO) is an important substrate in aerobic fermentations and may be a

limiting substrate, since oxygen gas is sparingly soluble in water.
 At high cell concentrations, the rate of oxygen consumption may exceed the rate of oxygen
supply, leading to oxygen limitations.
 When oxygen is the rate-limiting factor, specific growth rate varies with dissolved-oxygen
concentration according to Monod equation, just like any other substrate-limited case.
 Above a critical oxygen concentration, the growth rate becomes independent of the
dissolved oxygen concentration (DO). (See figure showing variation of specific growth
rate with DO in a rich medium, i.e., no other substrate limitation.
 Oxygen is a growth rate-limiting factor when the DO level is below the critical DO level.
 In this case, another medium component (e.g. glucose, ammonium) becomes growth-extent
limiting.

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  For example, with Azotobacter vinelandii at a DO = 0.05 mg/L, the growth rate is about
50% of maximum even if a large amount of glucose is present. However, the maximum
amount of cells formed is not determined by the DO, as oxygen is continually resupplied.
If glucose were totally consumed, growth would cease even if DO = 0.05 mg/L. Thus, the
extent of growth (mass of cells formed) would depend on glucose, while the growth rate
for most of the culture period would depend on the value of DO.


For aerobic microorganisms:

For facultative microorganisms (can grow with or without oxygen):

µmax0 = maximum growth under anaerobic condition

For anaerobic organisms, there is no growth if oxygen is present.

The critical oxygen concentration is about 5% to 10% of the saturated DO concentration for
bacteria and yeast and about 10% to 50% of the saturated DO concentration for mold cultures,
depending on the pellet size of molds.

Saturated DO concentration in water at 25°C and 1 atm pressure is about 7 ppm. The presence of
dissolved salts and organics can alter the saturation value, while increasingly high temperatures
decrease the saturation value.

Oxygen is usually introduced to the fermentation broth by sparging air through the broth.

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 8. Oxygen transfer

Oxygen transfer from gas bubbles to cells is usually limited by oxygen transfer through the
liquid film surrounding the gas bubbles. The rate of oxygen transfer from the gas to liquid phase
is given by:

where kL is the oxygen transfer coefficient (m/h),

a is the gas-liquid interfacial area (m2/m3),
C* is saturated DO concentration (g/L),
CL is the actual DO concentration in the broth (g/L),
and the NO2 is the rate of oxygen transfer (g/L/h).

The rate of oxygen uptake is denoted as OUR:

where µO2 is the specific rate of oxygen consumption (g g-cells/h),

YFX/O2 is the yield factor on oxygen (g-cells/g-O2), and
X is cell concentration (g-cells/L).

When oxygen transfer is the rate-limiting step, the rate of oxygen consumption is equal to the
rate of oxygen transfer. If the maintenance requirement of O2 is negligible compared to growth,
then

In batch reactor with negligible medium volume loss (due to air sparging):

The maximum or saturation oxygen concentration is a function of temperature, oxygen pressure,

as well as medium compositions. Electrolytes have a strong effect on oxygen solubility and
transport.

where C0* is the oxygen saturation concentration in pure water (Table 11.3),
C* is the oxygen saturation concentration in the medium,
Hj is the oxygen solubility interaction constants (Table 11.4),
Zj is the ionic charge of ionic species j, and
Cj is the concentration of species j in the fermentation medium.

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 9. Effect of Temperature

Microorganisms have various ranges in temperature in which they thrive and where they have
their maximum growth. They can be classified according to these growth temperatures.

Optimum temperature is the temperature at which they grow at maximum specific growth rate.

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 10. Effect of pH

Hydrogen ion concentration affects the activity of enzyme. Thus, it also affects microbial growth
rate.

Optimal pH for growth may be different from that for product formation. Acceptable pH range
varies from optimum pH by 1 – 2 pH units.

Different organisms have different pH optima:

Bacteria: 3 – 8
Yeast: 3 – 6
Molds: 3 – 7
Plant cells: 5 – 6
Animal cells: 6.5 – 7.5

Many organisms can maintain intracellular pH at relatively constant level even when pH of its
environment fluctuates.  maintenance energy increases.
pH may be affected by the N-source:
o if ammonium as sole N source: H+ are released into the medium (consume ammonia) 
decrease in pH
o if nitrate: H+ is removed from the media (to reduce nitrate to ammonia)  increase in pH

APRollon ChE 233 BRD Lecture 2.7 How Cells Grow in Batch Culture Page 26 of 28
 11. Balance Equation for Batch Reactor (BR)

For a three material system (substrate, cell, product):

S  X + P
substrate cells product

dC Sl 1 1
Substrate balance: V  C SlV  q P C XlV Eqn. 2.8
dt YSl YSP

dC Xl
Biomass Balance: V  C XlV  k d C XlV Eqn. 2.9
dt

dC Pl
Product Balance: V  q P C XlV Eqn. 2.10
dt

mass
concentration substrate

product cell

time

 Initial rate of substrate consumption is slow as cell growth is in the lag phase.
 As substrate is consumed, product and cell mass are formed.
 Net cell growth ceases when substrate is depleted.
 Eventually cell mass will decrease as product mass is virtually constant.
 In environmental process in BR where the starting medium is not sterile (i.e. other
organisms, provided other growth factors are sufficiently available)

APRollon ChE 233 BRD Lecture 2.7 How Cells Grow in Batch Culture Page 27 of 28
 o Fed-Batch Reactor (FBR)

BR: One cycle (lag, exponential, stationary, and death)  new cycle
 considerable downtime (time in which reactor is not productive)

CSTR: No considerable downtime; Not suited for some processes (example,

when idiophase is independent of the tropophase; more complex
operation)

FBR: To meet both requirement of more or less continuous operation and

continuously changing environment.
 When nutrients approach depletion, fresh nutrients are ‘fed’ to the reactor.
 The concentration of the nutrients added is so high that volume changes are
negligible. Or the concentration of the feed can be adjusted.

Example of FBR process:

 Landfill
 Fill and draw sedimentation tank

APRollon ChE 233 BRD Lecture 2.7 How Cells Grow in Batch Culture Page 28 of 28