BT 411

Course Title: Industrial Biotechnology

Fermentation
Rohit Kumar
29-Aug-2024
 Syllabus

S.No Contents/Syllabus
01 Introduction to industrial bioprocess: Fermentation, Basic concepts of
Upstream and Downstream processes

02 Production of primary metabolites: Organic acids, Amino acids and

alcohols.

03 Production of secondary metabolites: Antibiotics, Vitamins and Steroids

04 Production of enzymes and other bioproducts: Production of Industrial

Enzymes, Biopesticides, Biofertilizers, Biopreservatives, Biopolymers
Biodiesel. Cheese, Beer, SCP & Mushroom culture, Bioremediation.
05 Production of modern biotechnology products: Production of
recombinant proteins, vaccines.
 Fermentation
Fermentation:- Fermentation is an anaerobic process where, even though oxygen is not
available, energy can be released from glucose. In yeast cells, fermentation takes place, and
a process of fermentation takes place in bacteria and animal muscle cells.

Fermentation is the process involving the biochemical activity of organisms, during their
growth, development, reproduction, even senescence and death. Fermentation technology is
the use of organisms to produce food, pharmaceuticals and alcoholic beverages on a large
scale industrial basis.

The basic principle involved in the industrial fermentation technology is that organisms are
grown under suitable conditions, by providing raw materials meeting all the necessary
requirements such as carbon, nitrogen, salts, trace elements and vitamins.

The end products formed as a result of their metabolism during their life span are released
into the media, which are extracted for use by human being and that have a high
commercial value. The major products of fermentation technology produced economically
on a large-scale industrial basis are wine, beer, cider, vinegar, ethanol, cheese, hormones,
antibiotics, complete proteins, enzymes and other useful products.
 Fermentation Methodology
Fermentation Methodology:
Fermentation process is carried out in a container called the fermentor or bioreactor. The
design and nature of the fermentor varies depending upon the type of fermentation
carried out.
Invariably all the fermentors have facilities to measure some of the fermentation
parameters like temperature, pressure, pH, elapsed fermentation time, liquid level, mass
etc.
The different types of fermentors are:
1. External recycle airlift fermentor—for producing bacterial biomass, with methanol as
substrate.
2. Internal recycle airlift fermentor—for producing yeast with oil as substrate.
3. Tubular tower fermentor—Used for making beer, wine, vinegar etc.
4. Nathan fermentor—used in brewing industry.
5. Stirred fermentor—used for making antibiotics.
 Types of Fermentation
The following points highlight the eight main types of fermentations.
The types are:-
1. Batch Fermentation
2. Continuous Fermentation
3. Fed Batch Fermentation
4. Anaerobic Fermentation
5. Aerobic Fermentation
6. Surface Fermentations
7. Submerged Fermentations
8. State Fermentation.
 Types of Fermentation
Type # 1. Batch Fermentation: 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. 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. The used
medium along with cells of microorganism and the product is drawn out from the fermenter.
When the desired product is formed in optimum quantities, the product is separated from the
microorganism and purified later on
There are three phases in the batch fermentation process - lag phase, exponential phase, and
stationary phase.
•In the lag phase, microbes adapt to the environment of the culture and
•In the exponential phase, the microbial cells grow rapidly and consume most of the nutrients
and the last phase is
•The stationary phase is when the growth of microbes stops due to the consumption of all
nutrients. It is the simplest type of all industrial fermentation.

The growth of a bacterial culture can be represented by a curve that consists of four
stages or phases:
Lag phase - growth and reproduction are just beginning
Log phase - reproduction is occurring at an exponential rate
Stationary phase - environmental surroundings and food supply cannot support any
It has both advantages and disadvantages which are detailed below:
(i) Merits:
(a) The possibility of contamination and mutation is very less.
(b) Simplicity of operation and reduced risk of contamination.

(ii) Demerits:
(a) For every fermentation process, the fermenter and other equipment are to be cleaned
and sterilized.
(b) Only fraction of each batch fermentation cycle is productive.
(c) It is useful in fermentation with high yield per unit substratum and cultures that can
tolerate initial high substrate concentration.
(d) It can be run in repeated mode with small portion of the previous batch left in the
fermenter for inoculum.
(e) Use of fermenter is increased by eliminating turn round time or down time.
(f) Running costs are greater for preparing and maintaining stock cultures.
(g) Increased, frequency of sterilization may also cause greater stress on instrumentation
and
probes.
(h) Fresh sterilized medium and pure culture are to be made for every fermentation
process.
Type # 2. Continuous Fermentation: It is a closed system of fermentation, run for indefinite
period. In this method, fresh nutrient medium is added continuously or intermittently to the
fermenter and equivalent amount of used medium with microorganisms is withdrawn
continuously or intermittently for the recovery of cells or fermentation products. As a result,
volume of the medium and concentration of nutrients at optimum level are being maintained.
This has been operated in an automatic manner.

The continuous fermenter has its maximum use that take long time to reach high productivity,
reduces down time and lowers the operating costs. In continuous mode, starting medium and
inoculum are added to the fermenter. After the culture is grown the fermenter is fed with
nutrients and broth is withdrawn at the same rate maintaining a constant volume of broth in
the fermenter. In continuous mode with cell cycle, the cell mass is returned to the fermenter
using micro filtrations with bacteria or screens with fungal mycelium.
Fed-Batch Fermentation
 It is a modification of batch fermentation.

 In this nutrition is added aseptically and the amount of liquid culture in the
bioreactor increases as the culture is added systematically.

 It is a type of semi-open system.

 It yields a better result than batch fermentation.

 After consumption of early substrate continuous and constant nutrition is added.

 Types of Fermentation
Type # 5. Aerobic Fermentation: A fermentation process carried out in the presence of oxygen
is called as aerobic fermentation. In most of the commercial processes and majority of the
products of human utility are produced by this type of fermentation. Fermentation can be
surface culture or static and submerged.

Type # 6. Surface Fermentations: Surface fermentations are those where the substratum may
be solid or liquid. The organism grows on the substratum and draws the nutrients from the
substratum. These types of fermentations are desirable where the products are based on
sporulation. But it has several disadvantages such as it exposes the organism to unequal
conditions, both oxygen and nutrients.

Type # 7. Submerged Fermentations: Submerged Fermentations are those in which the

nutrient substratum is liquid and the organism grows inside the substratum. The culture
conditions are made uniform with the help of spargers and impeller blades. Most of the
industrial fermentations are of this type. The substratum which is in a liquid state and such
medium is also called as broth.

Type # 8. Solid Substrate/State Fermentation: Solid state (substratum) fermentation (SSF) is

generally defined as the growth of the microorganism on moist solid materials in the absence or
near the absence of free water. In recent years SSF has shown much promise in the
development of several bioprocesses and products, SSF has been ambiguously used as solid-
state fermentation or solid-substrate fermentation.