Outline: Fed-Batch Operation

Outline
Fed-batch operation
Models Advantages Disadvantages Example: penicillin production Advantages Disadvantages
Perfusion culture
Fed-Batch Operation
In fed-batch culture, nutrients are continuously or semi-continuously added to a system, while effluent is removed discontinuously
Usually used to overcome substrate inhibition or

catabolite repression
Recall that in batch culture the concentration of biomass at a certain time is given by:
X = X0 +Y
M X /S
(S0 S )
(9.27)
Schematic of a Fed-Batch Culture

1) 2)
3)
Fed-Batch Culture
When biomass concentration in a fed-batch reactor reaches its maximum (Xm), the substrate concentration is very low (S<<S0), and also X>>X0, therefore: M
X m = YX S S 0
If, for a fed batch reactor containing some cells (inoculum), a nutrient feed is started at a flow rate of F, with a substrate concentration of S0, the total amount of biomass in the vessel is Xt=VX, where V is the culture volume at time t, and Xt is the biomass at time t
Fed-Batch Culture
The rate of increase of culture volume in a fed batch reactor is: dV
dt
Integrating:
=F
(9.28)
V = V0 + Ft
(9.29)
The biomass concentration in the vessel at any time is: t
X=X V
(9.30)
Fed-Batch Culture
The rate of change in biomass concentration is:
dX V dX t dt X t (dV dt ) = 2 dt V
Since dXt/dt=netXt, dV/dt=F, and F/V=D, eq. 9.31 becomes:
(9.31)
dX = ( net D )X dt
(9.32)
Fed-Batch Culture
When the substrate is totally consumed, S0, and X=Xm
At this point, dX/dt=0, and the system is at quasi-steady state (nutrient consumption rate is nearly equal to nutrient feed rate), therefore: (9.33)
net = D
If maintenance energy can be neglected,
net
S = m Ks + S
and
Ks D S m D
(9.34)
Fed-Batch Culture
The balance on the rate-limiting substrate without maintenance energy is:
net X dS = FS 0 M dt YX S
t
(9.36)
Where St is the total amount of the rate-limiting substrate in the culture, and S0 is the concentration of S in the feedstream At quasi-steady state, Xt=VXm, and essentially all substrate is consumed, therefore:
dX t dV = Xm dt dt
M = X m F = FYX S S 0
(9.37)
Fed-Batch Culture
Integration of eq. 9.37 from t=0 to t, with the initial biomass concentration in the reactor t being X 0 yields:
X = X + FY
t t 0
M X S
S 0t
(9.38)
That is, the total amount of cells in the culture
increases linearly with time Dilution rate and net decrease with time Since net=D at quasi-steady state, the growth rate is controlled by the dilution factor
Fed-Batch Culture
For product formation in a fed-batch reactor, at quasi-steady state (S<<S0): P YP S S 0 (9.41) Or the potential product output is:
When the specific rate of product formation (qp) is constant:
dP = qp X t dt
(9.42)
FP YP S S 0 F
(9.43)
Where Pt is the total amount of product in culture
Fed-Batch Culture
Substituting Xt=(V0+Ft)Xm into eq. 9.41 yields:
Ft P = P + q p X m V0 + t 2
t t 0
(9.42)
Integration of eq. 9.42 yields:

t
Eq. 9.43 can be written in terms of product:
dP = q p X m (V0 + Ft ) dt
(9.43)
V0 V0 Dt P = P0 + q p X m + t V 2 V
(9.44)
Fed-Batch Culture at QuasiSteady State

(a) Variation of culture volume, V, specific growth rate, , cell, X, and substrate, S, concentration with time at quasi-steady state (b) Variation of product concentration, P, with time at quasi-steady state in a single cycle of a fed-batch culture (a)
Example 9.3
Solution
Advantages of Fed-Batch Culture

Production of high cell densities due to extension of working time (particularly important in the production of growth-associated products) Controlled conditions for the provision of substrates during the fermentation Control over the production of by-products, or catabolite repression effects, due to limited provision of only those substrates solely required for product formation Allows the replacement of water lost via evaporation Alternative mode of operation for fermentations involving bioremediation of toxic substrates (cells can only metabolize a certain quantity at a time), or low solubility compounds No additional special pieces of equipment are required to convert from batch to fed-batch operation
Disadvantages of Fed-Batch Culture

Requires previous analysis of the microorganism, its requirements, and an understanding of its physiology with respect to productivity Requires a substantial amount of operator skill for set-up, definition and development of the process In a cyclic fed-batch culture, care must be taken in the design of the process to ensure that toxins do not accumulate to inhibitory levels, and that nutrients other than those incorporated into the feed medium do not become limiting
Also, if many cycles are run, the accumulation of nonproducing or low-producing mutants may result
Fed Batch Culture: Penicillin Production

Fermentation is divided in two phases - the rapid-growth phase during which the culture grows at the maximum specific growth rate, and the slow-growth phase in which penicillin is produced During the rapid-growth phase:
An excess of glucose causes an accumulation of acids, and a biomass oxygen demand greater than the aeration capacity of the fermentor Glucose starvation may result in the organic nitrogen in the medium being used as a carbon source, resulting in a high pH and inadequate biomass formation Feed rates should be designed to limit the growth rate and oxygen consumption, such that a high rate of penicillin synthesis is achieved, and sufficient dissolved oxygen is available in the medium
During the production (slow-growth) phase:
Perfusion Culture
Perfusion systems are most often used for animal cell culture The basic characteristics are constant medium flow, cell retention, and, in some cases, selective removal of dead cells Cell retention is usually achieved by membranes or screens, or by a centrifuge capable of selective cell removal
When a membrane is used, the system has some characteristics of an immobilized cell system
Schematic of a Perfusion System
Advantages of Perfusion Culture

Potential removal of cellular debris and inhibitory by-products Removal of enzymes (e.g. proteases) released by dead cells that may destroy or damage product Shorter exposure time of product to potentially harsh production conditions (e.g. high or low pH) High per-unit volumetric productivity due to high cell density and metabolism Essentially constant environment
Disadvantages of Perfusion Culture

A large amount of medium is typically used
This is expensive, due to the high costs of raw materials for the medium, in addition to the costs of preparing and sterilizing the medium
Nutrients in the medium are less completely utilized than in batch or fed-batch systems Costs for waste treatment increase Must consider the trade-off of improved product quality and reactor productivity with the associated increased expense

Outline: Fed-Batch Operation

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Outline: Fed-Batch Operation

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Outline

Models Advantages Disadvantages Example: penicillin production Advantages Disadvantages

Usually used to overcome substrate inhibition or

Schematic of a Fed-Batch Culture

The biomass concentration in the vessel at any time is: t

If maintenance energy can be neglected,

That is, the total amount of cells in the culture

When the specific rate of product formation (qp) is constant:

Where Pt is the total amount of product in culture

Integration of eq. 9.42 yields:

Eq. 9.43 can be written in terms of product:

Fed-Batch Culture at QuasiSteady State

Advantages of Fed-Batch Culture

Disadvantages of Fed-Batch Culture

Fed Batch Culture: Penicillin Production

During the production (slow-growth) phase:

Schematic of a Perfusion System

Advantages of Perfusion Culture

Disadvantages of Perfusion Culture

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