Biokinetics: Cellular Batch Kinetics

Batch Growth

Cellular kinetics is largely dictated by the growth of cells, therefore the way in which a
microbe grows under a batch setting is critical to understanding cellular batch kinetics.
Growth is the critical response to a microorganism’s physiochemical environment. Cells
growth depends on various chemical, physical, and nutritional conditions.
Nutrients are essential and are used for:
i. Energy Production
ii. Biosynthesis
iii. Product Formation
Nutrient utilization can be described as follow

∑ S+ X → ∑ P+nX
Where S(Substrate), X(Biomass), and P (Cellular Product)
As we can see from the equation Cellular growth is an excellent example of an auto catalytic
reaction.
Net cellular growth can be characterized as follow:
1 dX
μnet = where X is∈ ( g/l )∧μnet is∈( h−1)(1)
X dt
μnet =μ g−k d where μ g growth rate∧k d death rate(2)

Growth Phases and Kinetics in Batch Culture

For a culture use for the inoculation of a liquid nutrient medium. The organisms take up
nutrients selectively with time to convert them into more biomass.
This is usually done in 5 phases in a batch setup.
Lag Phase

The first phase in cellular kinetics is a period of adaption in which the inoculate creates new
enzymes and repression of others that will not be needed for growth, the new enzymes once
created are use for the breakdown of the nutrients present to be converted into new biomass.
The length of the lag phase depends on several factors:
i. Concentration of nutrients and growth factors
ii. Age of inoculum
iii. Size of inoculum

Exponential Growth Phase

Once the lag phase is over it is followed by rapid growth. This is also a period of Balance
growth, which means all the cells parts are created at same rate.
In the exponential phase the cells follow first order kinetics:

dX
=μnet X
dt
X
ln =μnet t∨ X= X o e μ t (3)
net

Xo

Deceleration Phase

This Phase Begins due to a depletion of an important nutrient or accumulation of a toxic-by-

products.

Stationary Phase

When the net growth rate is a zero i.e., specific cell growth and death rates are equal is when
the stationary phase begins also secondary products are produce at this point.

Death Phase

Following the stationary phase as more materials are used or more toxic-by-products are
accumulated, more cells begin to die, hence increasing the specific death rate while on the
other hand the cell growth rate decreases.
The death rate for cells also follow first-order kinetics as follow:
dN
=−k d N∨N =N o e−k t Where N is cell number density ( 4)
d

dt
Yield Coefficients and Cellular Product Formation

Growth Kinetics can be defined even better by use of stoichiometrically related parameters
known as yield coefficients.
e.g., The growth yield in fermentation is
−∆ X
Y X= (5)
s
∆S

This yield coefficient however is not a true constant and therefore it is referred to as an
apparent growth yield or an observed yield, the reason why it can’t be considered as a
constant is due to the fact that the conditions of the culture can change with time which leads
to the uptake and use for substrates by the cells to also change therefore the apparent yield
will be different at different points of time in a fermenter.
Yield coefficients can also be based on other things such as product formation:
−∆ P
Y P= (6)
s
∆S

Yield coefficients for aerobic systems are usually more efficient than anaerobic systems and
therefore are usually much larger for a particular substrate.

Microbial Products

Cellular products grow in different ways in relation to cellular growth:

i. Growth associated products: Product are produced simultaneously with cellular
growth and the product formation rate is directly proportional to specific growth rate.

1 dP
q p= =Y P μ g (7)
X dt x

ii. Non growth associated products: Product that are form during the stationary phase,
the specific rate for the product formation is constant in this case.

q p=β=constant

iii. Mixed growth associated: Takes place between the slow growth and the stationary
phase. The rate of formation is as follow:

q p=αμ+ β
Quantifying Growth Kinetics

If we are to obtain a complete description of growth kinetics, we must incorporate the

structured nature of each cell present as well as the segregation of the culture into individual
units which differ from each other.
There for growth models can be:
i. Structured and Segregated (Most Complex)
ii. Unstructured and Segregated
iii. Structured and Nonsegregated
iv. Unstructured and Nonsegregated (Most Simple)
The model which is the simplest in describing the desired system adequately should be the
one chosen. However, for discussion of quantifying the kinetics unstructured and
nonsegregated will only be considered.

Unstructured Nonsegregated Models for Determination of Specific Growth

Rate

Substrate – Limited Growth

Monod Equation the simplest model assumes a single substrate is growth – limiting. The
kinetics are like Langmuir Hinshelwood as well as Michaelis – Menten kinetics for enzymes.
Note that unet =μ g for negligible cell death.
μm S
μ g= (8)
ks + S

Where ks is the saturation constant for substrate (S) and μm max specific growth rate
However, this model is often not very applicable since it can only be used when growth is
slow and cell concentration is low.
For rapid consumption and high concentration levels, toxic by-products become more
important to the kinetics and the specific growth equation takes the following form:
μm S
μ g= (9)
k so S o + S

Where kso is a dimensional constant and So is the initial substrate concentration

Growth Inhibition Models

Inhibition can be cause either by high concentrations of a substrate or high concentrations of

toxic – by-products. The inhibition can take two forms competitive and non-competitive.
For Substrates
μm
μ g= Noncompetitive(10)
k S
[ ][ ]
1+ s + 1+
S kI

μm S
μ g= Competitive(11)
S
S+ k s 1+
[ ]
kI

For Products
μm
μ g= Noncompetitive(12)
k P
[ ][ ]
1+ s + 1+
S kP

μm S
μ g= Competitive( 13)
P
S+ k s 1+
[ ]
kP

Where kI is saturation constant for the substrate acting as and inhibitor and kp is the saturation
constant of the inhibiting product.

Logistic Equations

Logistic equations are used as a way of describing growth in terms of the carrying capacity.
The general approach is the use of equations in which the specific growth rate is related to the
amount of unused carrying capacity.
X
μ g=k 1−( X∞ )
(14)

Combining equation 14 with 1 yield:

dX X
dt
=kX 1−
X∞(( 15) )
The following equations can be used to plot a logarithmic plot for the determination of the
carrying-capacity coefficient k:

1 dX X́
log
X dt
=logk + log 1−
X∞ (
(16) )
Where
Where X́ the average growth at time t and X∞ is the max possible growth of cells

Environmental Impact on Kinetics

Several environmental conditions have an impact on the patterns of cellular growth and
production formation.
i. Temperature
ii. pH
iii. Dissolved -Oxygen Concentration

Temperature

All microbes have a optima temp which they perform best at, above this point growth
decreases and then thermal death occurs, the following equations describe the kinetics above
the optima temperature.
dN
=( μ R−k d ) N
dt
− Ea −E d
RT
μ R= A e k d = A e RT

Temperature also affects product formation but optima temp for product formation may differ
from growth.

pH

pH has an impact on pH activity of the enzymes produced by microbes, therefore it has and
impact on growth rate. Different microbes also have different pH optima in which they
perform best at, generally:
i. Bacteria pH = 3 to 8
ii. Yeast pH = 3 to 6
iii. Molds pH = 3 to 7
iv. Animal Cells pH = 6.5 to 7.5

Dissolved Oxygen (DO)

For aerobic fermentation oxygen is of much importance however since water is sparingly
soluble oxygen transfer is usually a rate limiting step. The rate in which oxygen is transfer
from air to the broth is as follow:
N O =k L a ( C ¿−C L )=OTR
2
While for the uptake rate:
μg X
OUR=qO X =
2
YX
O2