5/5/2025

S Brain Storming Questions:

What is fermentation ?

What is industrial fermentation?

 What are fermenter, reactor and bioreactor (s)?

 What are the processes in the fermentation?

 What is fermentor configuration?

FERMENTATION: AN ANCIENT TRADITION

• Fermentation has been known and practiced by humankind

since prehistoric times which dates back to 3000 BC

• For example, the productions of bread, beer, vinegar, yogurt,

cheese, and wine were started in ancient Egypt

▪ Fermentation process is the oldest technology of all biotechnological

process
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Definitions For Fermentation

▪ The most common meaning of fermentation is the conversion of a

sugar into an organic acid or an alcohol

C6H12O6 → 2C2H5OH + 2CO2

▪ However, the term “fermentation” is also used in a broader sense

for the intentional use of microorganisms to make products useful
to humans on an industrial scale

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▪ The word ‘fermentation’ origin from the Latin verb ‘fevere’,

which means to boil
▪ Fermentation has three different meanings:
▪ 1st:- relates to microbial physiology, defined as the type of
metabolism of a carbon source in which energy is generated by
substrate level phosphorylation
▪ 2nd:- in terms of industrial microbiology, is any process in
which microorganisms are grown on a large scale,
▪ 3rd:- concerns food, is the process of converting carbohydrates
to alcohol or organic acids using microorganisms yeasts or
bacteria under anaerobic conditions
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▪ The term fermentation is used to indicate microbial cell

propagation and generation of products under either aerobic,
microaerobic, or anaerobic condition

▪ 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

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What Is Industrial Fermentation?

▪ Is the intentional use of fermentation technology for the large

production of microbial biomass or their metabolites

▪ Fermented products have immense use in food, medicine, and

other industries

▪ Modern industrial fermentation units use genetically modified

microbes for the rapid production of desired metabolites

Where fermentation takes place?

▪ Fermentation is carried out in vessels known as fermentor

▪ A fermentor provides controlled environment to the organisms

for the production of desired product

▪ The fermentor is also termed as bioreactor if plant, animal or

bacterial cells are used to make the product

▪ Fermenter prevents the entry and growth of contaminating

microbes from outside

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How To Achieving Sterility

➢ Physical Methods

▪ Asepsis

▪ Filtration

▪ Heat

▪ Radiations

➢ Chemical Methods

➢ Gaseous Sterility:-Ethylene oxide

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The Sterilization Of The Fermenters And Its

Accessories

 The fermentor itself, unless sterilized, is a source of

contamination

 Steam is used to sterilize the medium in situ(cip) in the

fermenter

 Crevices and rough edges are avoided in the construction of

fermentors.

 Probes used for monitoring fermentor activities, namely probes

for dissolved oxygen, CO2, pH, foam, etc., should also be
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Fermentation Process

▪ Process in industrial biotechnology is divided into a number of sections:

🖑 In-bound logistics:- the delivery and storage of raw materials

🖑 Upstream processing:- the processing of raw materials for the

fermentation

🖑 Fermentation:-where the major conversion occurs

🖑 Downstream processing:- the purification and concentrating of the raw

product(s)

🖑 Out-bound logistics:- the final packaging, storage and delivery of the

purified product(s)

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➢The entire process can be divided in three stages:-

1. Upstream processing
2. Fermentation process
3. Downstream process

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Upstream Processing

▪ Involves preparation of liquid medium, separation of particulate

and inhibitory chemicals from the medium, sterilization, air
purification etc.
▪ Formulation of media to be used in culturing the process
▪ Sterilization of the medium, fermenters and ancillary equipment
▪ Production of an active, pure culture in sufficient quantity to
inoculate the production vessel

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Fermentation Processing

▪ Involves the conversion of substrates to desired product with

the help of biological agents such as microorganisms

Substrate

Product

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Downstream Processing
➢ The processes that follows fermentation:-

▪ The growth of the organism in the production fermenter

under optimum conditions for product formation-
fermentation

▪ Cell harvesting

▪ Cell disruption

▪ Product purification from cell extracts or the growth medium

▪ The disposal of effluents produced by the process-waste

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Scale Up Fermentation

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Fermentation volume and product growth

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Fermentation raw materials

Upstream Processes Source of carbon, nitrogen, phosphorus, Sulphur,
mineral elements, trace elements, growth factors,
Microorganism water, etc. (availability, cost, stability, and
pretreatment and sterilization requirements)

Initial Isolation
Media development

Strain Improvement Propagation medium Maintenance medium

Production Medium
Production Strain Starter Culture
Constraints: nutritional requirements, propagation
metabolic controls, shear sensitivity, +/_
temperature optima, morphology, pH, O2 and Oxygen
CO2 effects and requirements, genetic pH control
stability, metabolic byproducts, viscosity Antifoam
effects Cooling/Heating
Supported or suspended growth, fermentor
type, striking mechanism, size, geometry,
mode of operation and automation
Fermentation 37

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The Range Of Fermentation Process

▪ There are five major groups of commercially important

fermentation:
i. Those that produce microbial cells (cell biomass)
ii. Those that produce microbial enzymes
iii. Those that produce microbial metabolites
iv. Those that produce microbial recombinant products
v. Those that modify a compound which are added to
the fermentation –perform biotransformation
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Mode And Types Of Fermentation

A. Modes of Fermentation

▪ There are general three modes of fermentation

operation are there:

▪ Batch Fermentation

▪ Continuous Fermentation and,

▪ Fed-batch Fermentation

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i. Batch of Fermentation

▪ Batch culture technique is also called closed system cultivation

▪ At first, nutrient solution is prepared, filled to the fermenter

and it is inoculated with inoculum

▪ Then nothing is added in the fermentation tank except aeration

▪ In the batch culture, neither fresh medium is added nor used up

media removed from culture vessel during fermentation time

▪ Therefore, the volume of the culture remains the same

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▪ However, concentration of nutrition decrease continuously

▪ Accumulation of wastes keep increasing as a time goes

I Cells are growth in a batch reactor, goes through a series of

stages:

❖ Lag phase

❖ Exponential phase

❖ Stationary phase

❖ Death phase

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Advantages of Batch Fermentation

I Batch culture systems provide a number of advantages :

❖ Reduced risk of contamination or cell mutation

❖ Lower capital investment when compared to continuous

processes for the same bioreactor volume

❖ More flexibility with varying product/biological systems

❖ Higher raw material conversion levels

❖ Processes using one reactor to make various products

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Disadvantages of Batch Fermentation

I Batch culture systems disadvantages might be:

❖ The limitation and depletion of the substrate –toxins form
there

❖ the growth pattern may reach the death phase quickly in

an old culture

❖ Inhibition is another factor affecting the bioprocess

❖ process is highly labor intensive for downtime and

cleaning
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ii. Continuous Fermentation

▪ It represents an open system, in which fresh sterile medium is
added continuously in the vessel and used up media with cell
culture is removed continuously at the same rate

▪ the volume and cell density remain same in the cultivation vessel

▪ In this case, cells grow continuously in their log phase

▪ This type of growth termed as “steady state growth”

▪ The cell density in the culture remains constant and it is achieved

by maintaining constant dilution and flow rate

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Microbial growth in Chemostat

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Advantages of Continuous Fermentation

I Continuous fermentation provide a number of advantages :

▪ Growth rate is higher as nutrients are continuously added
to the fermentation tank
▪ Continuous cultures are more efficient as the fermenter
operates continuously
▪ Very useful for processes involving the production of
primary metabolites

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Disadvantages of Continuous Fermentation

I Continuous fermentation also has some limitations such as:

▪ Set up is more difficult, the maintenance of required
growing conditions can be difficult to achieve
▪ If contamination occurs, huge volumes of product may be
lost

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iii. Fed-batch Fermentation

▪ A semi-closed system of cultivation in which one or more nutrients
are gradually added to the fermenter during fermentation time
without removing used up media

▪ So, the volume of the culture broth in the vessel increases

continuously

▪ Such essential nutrient for the fermentation, initially kept in lower

concentration and added slowly and continuously during the course
of fermentation

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➢The main advantages of fed-batch over batch cultures are:

▪ the possibility to prolong product synthesis

▪ the ability to achieve higher cell densities and thus increase

the amount of the product

▪ the capacity to enhance yield or productivity by controlled

sequential addition of nutrients,

▪ the feature of prolonged productive cultivation over the

“unprofitable periods”

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➢Disadvantages include:

▪ Lower productivity levels due to time for filling, heating,

sterilization, cooling, emptying and cleaning the reactor

▪ Higher costs in labor and/or dynamic process control for the

process

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B. Types of Fermentation

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Industrial Fermenter /Bioreactor

▪ Bioreactors are vessels or tanks in which whole cells or cell-

free enzymes transform raw materials into biochemical
products and/or less undesirable by-products

▪ The microbial cell itself is a miniature bioreactor; other

examples include shake flasks, Petri dishes, and industrial
fermentors

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Bioreactor Design Criteria

• To design a bioreactor, some objectives have to be defined

• The decisions made in the design of the bioreactor might have

a significant impact on overall process performance

• The bioreactor is the heart of any biochemical process in

which enzymes, microbial, mammalian or plant cell systems
are used for manufacture of a wide range of useful biological
products

• The performance of any bioreactor depends on many

functions such as:
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Biomass concentration Heat removal

Nutrient supply Aeration
Sterile conditions Correct shear conditions
Product removal Metabolisms/microbial activities
Effective agitations Implication of bioreactor design on
Product inhibition downstream product separation
Aseptic equipment design Capital and operating costs of the
Control of bioreactor bioreactor
environment Potential for bioreactor scale-up

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Parts of Fermenter /Bioreactor

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What are the design criteria…?

Microbiological and biochemical characteristics of the cell

systems (Microbial, mammalian, plant cell)

Hydrodynamic characteristics of the bioreactor

Mass and heat characteristics of the bioreactor

Kinetics of cell growth and product formation

Genetic stability characteristics of the cell system

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Fermentors Configuration

Different fermentors configuration are based on:

▪ Shape or configuration of vessel(Fermentor)
▪ Aerated or anaerobic
▪ They are batch or continuous
The construction and design of a fermentor are the province of the
engineer
Construction is as will help the biotechnologist or microbiologist
understand and utilize it efficiently
Its sampling point and others related to fermentation approved by
biotechnologist/Microbiologist
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Bioreactor Configuration

⚫It affects the liquid flow pattern

⚫The performance of a fermentation in

terms of yield and production rate

❑ Bioreactors can be classified into different types

based on many features

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1. Based on the presence or absence of oxygen and requirement

of stirring bioreactors can be conveniently classified into three
major types
• about 70% of bioreactor
• anaerobic processes, in which
Non-stirred, non-aerated system wine, beer, cheese produced

Non-stirred, aerated system • 10% of current bioreactor

• aerobic solid state fermentation
processes, production of food enzymes
Stirred and aerated systems
• 20% of the bioreactor in the industrial application
• aerobic submerged fermentation processes,
production of citric acid and penicillin

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2. Based on mode of operation, the bioreactors can be classified

into three types:

❖Batch reactors

❖Fed batch reactors

❖Continuous reactors

3. Based on the method of growing of microbes, bioreactors can be

either:

❖Suspended

❖Immobilized (Petri dish, Fibrous bed, Packed bed,

Membrane, and others… 73

4. On the basis of the microbial agent used, the bioreactors

can be classified into:

➢ Those based on living cells (either microbes, plant and

animal cells)

➢Which employ enzymes (biochemically active

substance –biocatalysts)

5. Based on the process requirements, bioreactors can be

classified into:-

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I. Aerobic

II. Anaerobic

III. Solid state

IV. Immobilized

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I. Aerobic Bioreactor

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a) Stirred Tank Bioreactor

▪ The most common type of bioreactor used in industry

▪ A draught is fitted which provides a defined circulation pattern

▪The stirrer can be either at the top or bottom

of the reactor
▪The dimensions of the reactor depend on the
amount of heat to be removed from the vessel
▪Baffles in the center of the tank prevent
formation of vortex and effective mixing
of the ingredients

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b) Airlift Bioreactor

• The bioreactor consists of two concentric

cylinders – creating outer and inner
chambers
• The bottom of inner chamber carries a sintered steel ring
though which 5% CO2/O2-8% in air is bubbled 80

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c) Bubble Column Bioreactor

• In this type of bioreactor, the air is introduced at the base of the column
through perforated pipes or plates

• The vessel used for bubble column bioreactors is

usually cylindrical with an aspect ratio of 4-6 (h/d)

• For the production of baker’s yeast, beer

and vinegar, bubble column fermenters
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d) The fluidized bed fermenter:

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Generally

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Next Will Continuous …

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