Industrial Biochemistry

AIMMSCR, 4th Semester, B.Sc.(H) Biochemistry

Types of Bioreactors
Fermenter or bioreactor
• A biorector is a device in which the organisms are cultivated and motivated to form
a desired product

• Closed vessel or containment designed to give a right environment for optimal

growth and metabolic activity of the organism

• Fermenter: for microbes/ Bioreactor : for eukaryotic cells

• Size variable ranging from 20-250 million litres or more.

• Large scale production (10-100L to1000-million L capacity)

• Helps to meet requirements of:

pH
temp
aeration
agitation
drain or overflow
control systems
sensors
cooling to achieve maximum microbial yield
Basic modes of operations of a fermenter

1. Batch culture

2. Continuous culture

3. Fed-batch culture
Batch culture

Batch fermentation refers to

• A batch fermentation is a closed culture system, because initial and
limited amount of sterilized nutrient medium is introduced into the
fermenter. The medium is inoculated with a suitable microorganism and
incubated for a definite period for fermentation to proceed under optimal
physiological conditions. Oxygen in the form of air, an antifoam agent
and acid or base, to control the pH, are being added during the course of
fermentation process

• During the course of incubation, the cells of the microorganism undergo

multiplication and pass through different phases of growth and
metabolism due to which there will be change in the composition of
culture medium, the biomass and metabolites. The fermentation is run
for a definite period or until the nutrients are exhausted. The culture
broth is harvested and the product is separated.
• In this mode of operation,
conditions are continuously
changing with time, and the
fermentor is an unsteady-state
system, although in a well-mixed
reactor, conditions are supposed to
be uniform throughout the reactor
at any instant time.
Batch fermentation may be used to produce biomass, primary metabolites and
secondary metabolites under cultural conditions supporting the fastest growth rate
and maximum growth would be used for biomass production. The exponential phase
of growth should be prolonged to get optimum yield of primary metabolite, while it
should be reduced to get optimum yield of secondary metabolites.

Advantages:

Possibility of Contamination is less

Simple operation
Disadvantages:
1 charging of the fermenter with fresh medium;
2 sterilization of the fermenter and medium;
3 inoculation of the fermenter;
4 production of microbial products;
5 harvesting of biomass and spent fermentation broth;
and finally
6 cleaning of the vessel.

The principal disadvantage of batch processing is the high proportion of unproductive

time (down-time) between batches
Continuous culture
Initially, continuous fermentations start as batch cultures, but exponential
growth can then be extended indefinitely, in theory, through the continuous
addition of fresh fermentation medium
Continuous culture is a technique involving feeding the microorganism used for
the fermentation with fresh nutrients and, at the same time, removing spent
medium plus cells from the system
 where D = dilution rate (per hour), F = flow (L/h) and
V = reactor volume (L).

The term D is the reciprocal of the mean residence time or hydraulic retention time

Therefore, under steadystate conditions the net biomass balance can be described as

Under steady-state conditions the rate of growth = rate of loss, hence dx/dt=0 and therefore

By changing the rate with which medium is added to the bioreactor the specific growth
rate of the microorganism can be easily controlled within limits.
If the dilution rate is increased above µmax, complete wash-out of the cells occurs, as the
cells have insufficient time to ‘double’ before being washed out of the reactor via the
overflow. The point at which this is just avoided is referred to as the critical dilution rate
(Dcrit).
3. Fed-batch processes

The fed-batch technique was originally devised by yeast producers in the early 1900s
to regulate the growth in batch culture of Saccharomyces

Figure 2.11. Fed-batch reactor

for a bioreaction.
A fed-batch is useful in achieving high concentration of products as a result of high
concentration of cells for a relative large span of time.

Two cases can be considered: the production of a growth associated product and the
production of a non-growth associated product. In the first case, it is desirable to extend the
growth phase as much as possible, minimizing the changes in the fermenter as far as specific
growth rate, production of the product of interest and avoiding the production of by-products.

For non-growth associated products, the fed-batch would be having two phases: a growth
phase in which the cells are grown to the required concentration and then a production phase
in which carbon source and other requirements for production are fed to the fermenter.
This case is also of particular interest for recombinant inducible systems: the cells are grown
to high concentrations and then induced to express the recombinant product

Fed-batch operation can extend the product formation phase and may overcome problems
associated with the use of repressive, rapidly metabolized,

. This method is also useful where a substrate causes viscosity problems or is toxic
at high concentrations.
The advantages of the fed-batch cultivation process are as
follows: It shortens fermentation time,
achieves high cell concentration,
increases productivity,
diminishes substrate inhibition or end-product inhibition, reduces
the viscosity of the culture broth,
reduces water loss by evaporation,
and gives a higher dissolved oxygen rate
Characteristics Batch culture Fed-batch culture Continuous culture
Cultivation system Closed type Semi-closed type Open type
Addition of fresh
nutrition No (limiting factor) Yes Yes
Internal environment Changes Changes Does not change
Lag, log, stationary and Lag, log , stationary and
Growth phase decline phase decline phase Lag and log phase
Log phase Shorter longer Longest and Continuous
Density of bacteria Change with time Change with time Remain same
Volume of culture Constant Increases Constant
Removal of product No No Yes
Removal of wastes No No Yes

Chance of contamination minimum Intermediate Maximum

Fermenter Size Large Medium Smaller
Labor Demand More Intermediate Less
Yield Low Intermediate High
Bioreactor is a vessel in which a biochemical process is carried out which involves
organisms or biochemically active substances derived from such organisms.

The main function of a fermenter is to provide a suitable environment in which an organism

can efficiently produce a target product that may be cell biomass, a metabolite or
bioconversion product.

It was during the First World War, a British scientist named Chain Weizmann (1914-1918)
developed a fermenter for the production of acetone.

FERMENTOR STAGE
 Lab scale

Pilot
scale
Industry scale
 Parts of a fermentor

The performance of any fermenter depends on many factors, but the key physical and
chemical parameters that must be controlled are agitation rate, oxygen transfer, pH,
temperature and foam production.

Aeration & agitation system

• Impeller
• Sparger
• Baffles
• Load cells
• Inlet & exit gas analyser
• pH meter
Sensors
• Flow cell
• Steam line
Basic design
Construction of Fermenters:

The criteria considered before selecting materials for constructions of a

fermenter are:

(a) The materials that have no effect of sterilization, and

(b) Its smooth internal finish – discouraging lodging of contamination. The
internal surface should be corrosion-resistant.

There are two types of such materials:

(i) Stainless steel, and
(ii) Glass which are used in fermenter.
 (i) Control of Temperature:

Heat is produced by microbial activity and mechanical agitation, then it is

sometimes necessary to remove it.

On the other hand, in certain processes extra heat is produced by using

thermostatically controlled water bath or by using internal heating coil or jacket
meant for water circulation.

Heat transfer is primarily achieved using

an outer jacket surrounding the internal
phase or via internal coils. No direct
contact exists between the cooling/
heating system and the fermentation
medium.
 (ii) Aeration and Agitation:
The main purpose of aeration and agitation is to provide oxygen required to the
metabolism of microorganisms.

The agitation should ensure a uniform suspension of microbial cells suspended in

nutrient medium. Agitation of suspended cell fermentations is performed in order
to mix the three phases within a fermenter

Mixing should produce homogeneous conditions and promote nutrient, gas and
heat transfer. Heat transfer is necessary during both sterilization and for
temperature maintenance during operation.

Efficient mixing is particularly important for oxygen transfer in aerobic

fermentations,

Maintenance of suitable shear conditions during the fermentation is very important

The mixing of nutrients and gaseous exchange within any fermenter is complex. It
is influenced by medium density and rheology, size and geometry of the vessel,
and the amount of power used in the system.
(a) The agitator (impeller) for mixing;
(b) Stirrer glands and bearings meant for aseptic sealing;
(c) Baffles for checking the vortex resulting into foaming;
(d) The sparger (aeration) meant for introducing air into the liquid.

(a) The Agitator (Impeller):

The size and position of the impeller in the vessel depends upon the size of the
fermenter. In tall vessels, more than one impeller is needed if adequate aeration
agitation is to be obtained. Ideally, the impeller should be 1/3 or 1/2 of the vessel
diameter (D) above the base of the vessel. The number of impeller may vary from
size to size to the vessel
 Impeller blades

The effectiveness of agitation depends upon the

design of the impeller blades,
speed of agitation and
the depth of liquid.
 Agitation

Pneumatic systems, such as airlift fermenters Hydrodynamic mechanisms use liquid

use the expansion of compressed gas to bring kinetic energy
about the mixing to mix the fermenter contents,
Baffles:
The baffles are normally incorporated into agitated vessel of all sizes to prevent a
vortex and to improve aeration efficiency. They are metal strips roughly one-
tenth of the vessel diameter and attached radially to the walls.
 Aeration
Some fermentations operate anaerobically, but the majority are aerobic and
require the provision of large quantities of normally sterile air or oxygen
To prevent the risk of contamination, gases introduced into
the fermenter should be passed through a sterile filter

A similar filter on the air exhaust system avoids environmental

contamination.

Sterile filtered air or oxygen normally enters the fermenter through a sparger
system. A sparger may be defined as a device for introducing air into the
liquid in a fermenter.

To promote aeration in stirred tanks, the sparger is usually located

directly below the agitator.

Sparger structure can affect the overall transfer of oxygen into the medium, as it
influences the size of the gas bubbles produced.
Three basic types of sparger have been used and may be described as the porous
sparger, the orifice sparger and the nozzle sparger.

Oxygen transfer is complex, as it involves a phase

change from its gaseous phase to the liquid phase, and is
influenced by the following factors.
1 the prevailing physical conditions; temperature, pressure
and surface area of air/oxygen bubbles;
2 the chemical composition of the medium;
3 the volume of gas introduced per unit reactor volume
per unit time;
4 the type of sparger system used to introduce air into
the fermenter;
5 the speed of agitation; or
6 a combination of these factors.

Challenge: Oxygen is only sparingly soluble in aqueous solution and the solubility
decreases as the temperature rises.
 Fermenter control and monitoring

Fermentation systems must be efficiently controlled in order to optimize

productivity and product yield, and ensure reproducibility.

primarily aeration, mixing, temperature, pH and foam control.

Control and maintenance at optimum levels inside the reactor is mediated by

sensors (electrodes),
Internal Sensors

External Sensors

All parameters (temperature, pH, pO2, air flow rate,

agitation,. pCO2, optical density, antifoam etc.) are
visible at a glance on a large LCD back light display .
The basic principle of control involves a sensory system linked to a control system and
feedback loop
 Types of Bioreactors

• Simple fermenters (batch and continuous)

• Fed batch fermenter
• Air-lift fermenter
• Cyclone column fermenter
• Tower fermenter
• Fluidized bed bioreactors
• Packed bed bioreactor
• Photobioreactor
• Bubble reactor
• Solid State fermenter
• Rotary drum Reactor
• Hollow fiber reactor
Aastha Aggarwal •Air-lift fermenter

Aryan Kandari
•Solid State fermenter
Eshika Jadon immobilized bioreactor
Krishna
•Fluidized bed bioreactors
Nishika Das
•Packed bed bioreactor
R Shweta
•Photobioreactor
Saloni Jain
•Bubble reactor
Sheen Razdan
Cyclone column fermenter
Riya Arora
•Rotary drum Reactor
Ritika Hollow fiber reactor
Shubhra •Tower fermenter